CORD-19-Sentences  

CORD-19:500c5ea0961b573ba5355d364ca94c00a0a26e4a JSONTXT 8 Projects

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Id Subject Object Predicate Lexical cue
TextSentencer_T1 0-153 Sentence denotes The dengue virus non-structural protein 1 (NS1) is secreted from mosquito 2 cells in association with the intracellular cholesterol transporter chaperone
TextSentencer_T1 0-153 Sentence denotes The dengue virus non-structural protein 1 (NS1) is secreted from mosquito 2 cells in association with the intracellular cholesterol transporter chaperone
TextSentencer_T2 155-163 Sentence denotes Abstract
TextSentencer_T2 155-163 Sentence denotes Abstract
TextSentencer_T3 164-179 Sentence denotes 11 12 Keywords:
TextSentencer_T3 164-179 Sentence denotes 11 12 Keywords:
TextSentencer_T4 180-451 Sentence denotes 13 dengue virus; zika virus; yellow fever virus; flavivirus; NS1; caveolin-1; chaperone caveolin 14 complex; viral protein trafficking; unconventional secretion; mosquito cells 15 16 17 ABSTRACT 18 Dengue virus (DENV) is a mosquito-borne virus of the family Flaviviridae.
TextSentencer_T4 180-451 Sentence denotes 13 dengue virus; zika virus; yellow fever virus; flavivirus; NS1; caveolin-1; chaperone caveolin 14 complex; viral protein trafficking; unconventional secretion; mosquito cells 15 16 17 ABSTRACT 18 Dengue virus (DENV) is a mosquito-borne virus of the family Flaviviridae.
TextSentencer_T5 452-575 Sentence denotes The RNA viral genome 19 encodes for a polyprotein that is co-translationally processed into three structural proteins and .
TextSentencer_T5 452-575 Sentence denotes The RNA viral genome 19 encodes for a polyprotein that is co-translationally processed into three structural proteins and .
TextSentencer_T6 576-633 Sentence denotes CC-BY 4.0 International license is made available under a
TextSentencer_T6 576-633 Sentence denotes CC-BY 4.0 International license is made available under a
TextSentencer_T7 634-724 Sentence denotes The copyright holder for this preprint (which was not peer-reviewed) is the author/funder.
TextSentencer_T7 634-724 Sentence denotes The copyright holder for this preprint (which was not peer-reviewed) is the author/funder.
TextSentencer_T8 725-818 Sentence denotes It . https://doi.org/10.1101/370932 doi: bioRxiv preprint 2 20 seven non-structural proteins.
TextSentencer_T8 725-818 Sentence denotes It . https://doi.org/10.1101/370932 doi: bioRxiv preprint 2 20 seven non-structural proteins.
TextSentencer_T9 819-965 Sentence denotes The non-structural protein 1 (NS1) is a multifunctional viral protein 21 actively secreted in vertebrate and mosquito cells during DENV infection.
TextSentencer_T9 819-965 Sentence denotes The non-structural protein 1 (NS1) is a multifunctional viral protein 21 actively secreted in vertebrate and mosquito cells during DENV infection.
TextSentencer_T10 966-1074 Sentence denotes In mosquito cells, NS1 22 is secreted in a caveolin-1 (CAV-1) dependent manner by an unconventional pathway.
TextSentencer_T10 966-1074 Sentence denotes In mosquito cells, NS1 22 is secreted in a caveolin-1 (CAV-1) dependent manner by an unconventional pathway.
TextSentencer_T11 1075-1265 Sentence denotes The 23 caveolin chaperone complex (CCC) is a cytoplasmic complex formed by caveolin-1 and the 24 chaperones FKBP52, Cy40 and CyA which is responsible for cholesterol traffic inside the cell.
TextSentencer_T11 1075-1265 Sentence denotes The 23 caveolin chaperone complex (CCC) is a cytoplasmic complex formed by caveolin-1 and the 24 chaperones FKBP52, Cy40 and CyA which is responsible for cholesterol traffic inside the cell.
TextSentencer_T12 1266-1460 Sentence denotes 25 In this work, we demonstrate that in infected mosquito cells, DENV NS1 is secreted by an early 26 and unconventional route that bypasses the Golgi apparatus in close association with the CCC.
TextSentencer_T12 1266-1460 Sentence denotes 25 In this work, we demonstrate that in infected mosquito cells, DENV NS1 is secreted by an early 26 and unconventional route that bypasses the Golgi apparatus in close association with the CCC.
TextSentencer_T13 1461-1693 Sentence denotes 27 Treatment of mosquito cells with classic secretion inhibitors such as brefeldin A, golgicide A and 28 Fli-06 showed no effect on NS1 secretion, but significant reductions in recombinant luciferase 29 secretion and virion release.
TextSentencer_T13 1461-1693 Sentence denotes 27 Treatment of mosquito cells with classic secretion inhibitors such as brefeldin A, golgicide A and 28 Fli-06 showed no effect on NS1 secretion, but significant reductions in recombinant luciferase 29 secretion and virion release.
TextSentencer_T14 1694-1879 Sentence denotes Silencing the expression of CAV1, FKBP52 with siRNAs or the 30 inhibition of CyA by cyclosporine A resulted in significant decrease in NS1 secretion without 31 affecting virion release.
TextSentencer_T14 1694-1879 Sentence denotes Silencing the expression of CAV1, FKBP52 with siRNAs or the 30 inhibition of CyA by cyclosporine A resulted in significant decrease in NS1 secretion without 31 affecting virion release.
TextSentencer_T15 1880-2075 Sentence denotes Using co-localization, co-inmunoprecipitation and proximity ligation 32 assays, NS1 was found to co-localize and interact with all the protein components of the CCC 33 in mosquito infected cells.
TextSentencer_T15 1880-2075 Sentence denotes Using co-localization, co-inmunoprecipitation and proximity ligation 32 assays, NS1 was found to co-localize and interact with all the protein components of the CCC 33 in mosquito infected cells.
TextSentencer_T16 2076-2163 Sentence denotes In addition, CAV-1 and FKBP52 expression was found augmented in 34 DENV infected cells.
TextSentencer_T16 2076-2163 Sentence denotes In addition, CAV-1 and FKBP52 expression was found augmented in 34 DENV infected cells.
TextSentencer_T17 2164-2321 Sentence denotes Finally, the treatment of ZIKV infected mosquito cells with brefeldin A and 35 golgicide A showed no effect on NS1 secretion, while affecting virion release.
TextSentencer_T17 2164-2321 Sentence denotes Finally, the treatment of ZIKV infected mosquito cells with brefeldin A and 35 golgicide A showed no effect on NS1 secretion, while affecting virion release.
TextSentencer_T18 2322-2402 Sentence denotes ZIKV NS1 was 36 also found to co-localize with CAV-1 in infected mosquito cells.
TextSentencer_T18 2322-2402 Sentence denotes ZIKV NS1 was 36 also found to co-localize with CAV-1 in infected mosquito cells.
TextSentencer_T19 2403-2518 Sentence denotes These results suggest that in 37 mosquito cells, ZIKV NS1 follows the same secretory pathway observed for DENV NS1.
TextSentencer_T19 2403-2518 Sentence denotes These results suggest that in 37 mosquito cells, ZIKV NS1 follows the same secretory pathway observed for DENV NS1.
TextSentencer_T20 2519-2646 Sentence denotes The 38 association of NS1 with the cholesterol transporter CCC agrees with the lipoprotein nature of 39 secreted hexameric NS1.
TextSentencer_T20 2519-2646 Sentence denotes The 38 association of NS1 with the cholesterol transporter CCC agrees with the lipoprotein nature of 39 secreted hexameric NS1.
TextSentencer_T21 2647-2669 Sentence denotes 40 41 AUTHOR SUMMARY .
TextSentencer_T21 2647-2669 Sentence denotes 40 41 AUTHOR SUMMARY .
TextSentencer_T22 2670-2818 Sentence denotes CC-BY 4.0 International license is made available under a The copyright holder for this preprint (which was not peer-reviewed) is the author/funder.
TextSentencer_T22 2670-2818 Sentence denotes CC-BY 4.0 International license is made available under a The copyright holder for this preprint (which was not peer-reviewed) is the author/funder.
TextSentencer_T23 2819-2955 Sentence denotes It . https://doi.org/10.1101/370932 doi: bioRxiv preprint 3 42 Dengue protein NS1 is secreted in infected mosquito and vertebrate cells.
TextSentencer_T23 2819-2955 Sentence denotes It . https://doi.org/10.1101/370932 doi: bioRxiv preprint 3 42 Dengue protein NS1 is secreted in infected mosquito and vertebrate cells.
TextSentencer_T24 2956-3022 Sentence denotes In humans, secreted 43 NS1 have been associated with pathogenesis.
TextSentencer_T24 2956-3022 Sentence denotes In humans, secreted 43 NS1 have been associated with pathogenesis.
TextSentencer_T25 3023-3125 Sentence denotes In mosquito cells, NS1 follows an unconventional 44 secretion pathway that is dependent on Caveolin-1.
TextSentencer_T25 3023-3125 Sentence denotes In mosquito cells, NS1 follows an unconventional 44 secretion pathway that is dependent on Caveolin-1.
TextSentencer_T26 3126-3230 Sentence denotes This work shows that in mosquito cells, NS1 45 secretion is associated to the chaperone caveolin complex
TextSentencer_T26 3126-3230 Sentence denotes This work shows that in mosquito cells, NS1 45 secretion is associated to the chaperone caveolin complex
TextSentencer_T27 3231-3343 Sentence denotes , a complex formed by caveolin-1 and 46 several chaperones, in charge of cholesterol transport within the cells.
TextSentencer_T27 3231-3343 Sentence denotes , a complex formed by caveolin-1 and 46 several chaperones, in charge of cholesterol transport within the cells.
TextSentencer_T28 3344-3517 Sentence denotes Reduction of the 47 expression or the activity of chaperone caveolin complex in mosquito infected cells, diminished 48 the secretion of NS1 without affecting virion release.
TextSentencer_T28 3344-3517 Sentence denotes Reduction of the 47 expression or the activity of chaperone caveolin complex in mosquito infected cells, diminished 48 the secretion of NS1 without affecting virion release.
TextSentencer_T29 3518-3631 Sentence denotes Direct interaction between NS1 and the 49 chaperone caveolin complex proteins was demonstrated by several assays.
TextSentencer_T29 3518-3631 Sentence denotes Direct interaction between NS1 and the 49 chaperone caveolin complex proteins was demonstrated by several assays.
TextSentencer_T30 3632-3847 Sentence denotes Moreover, 50 increased expression of the caveolin-1 and co-chaperone FKBP52 during dengue infection was 51 found, presumably in response to the higher requirements of these proteins during dengue virus 52 infection.
TextSentencer_T30 3632-3847 Sentence denotes Moreover, 50 increased expression of the caveolin-1 and co-chaperone FKBP52 during dengue infection was 51 found, presumably in response to the higher requirements of these proteins during dengue virus 52 infection.
TextSentencer_T31 3848-4039 Sentence denotes Results obtained with ZIKV infected mosquito cells suggest that also ZIKV NS1 is 53 released following an unconventional secretory route in association with the chaperone caveolin 54 complex.
TextSentencer_T31 3848-4039 Sentence denotes Results obtained with ZIKV infected mosquito cells suggest that also ZIKV NS1 is 53 released following an unconventional secretory route in association with the chaperone caveolin 54 complex.
TextSentencer_T32 4040-4098 Sentence denotes The functions of secreted NS1 within mosquito are unclear.
TextSentencer_T32 4040-4098 Sentence denotes The functions of secreted NS1 within mosquito are unclear.
TextSentencer_T33 4099-4299 Sentence denotes However, giving the 55 importance of the soluble NS1 in the vertebrate host, manipulation of the NS1 secretory route 56 may prove a valuable strategy for dengue mosquito control and patient treatment.
TextSentencer_T33 4099-4299 Sentence denotes However, giving the 55 importance of the soluble NS1 in the vertebrate host, manipulation of the NS1 secretory route 56 may prove a valuable strategy for dengue mosquito control and patient treatment.
TextSentencer_T34 4300-4442 Sentence denotes 57 58 59 4 64 infection and its public health burden are hard to estimate, but at least 100 countries are currently 65 endemic for dengue [2].
TextSentencer_T34 4300-4442 Sentence denotes 57 58 59 4 64 infection and its public health burden are hard to estimate, but at least 100 countries are currently 65 endemic for dengue [2].
TextSentencer_T35 4443-4502 Sentence denotes DENV has a single stranded RNA genome of positive polarity.
TextSentencer_T35 4443-4502 Sentence denotes DENV has a single stranded RNA genome of positive polarity.
TextSentencer_T36 4503-4694 Sentence denotes The 66 genome of DENV is translated into a single polyprotein and encodes for three structural (C, E, 67 prM/M) and seven non-structural (NS) (NS1, NS2A, NS2B, NS3, NS4A, NS4B, NS5) proteins.
TextSentencer_T36 4503-4694 Sentence denotes The 66 genome of DENV is translated into a single polyprotein and encodes for three structural (C, E, 67 prM/M) and seven non-structural (NS) (NS1, NS2A, NS2B, NS3, NS4A, NS4B, NS5) proteins.
TextSentencer_T37 4695-4802 Sentence denotes 68 The polyprotein is then cleaved by host and viral proteases to release individual viral proteins 69 [3].
TextSentencer_T37 4695-4802 Sentence denotes 68 The polyprotein is then cleaved by host and viral proteases to release individual viral proteins 69 [3].
TextSentencer_T38 4803-4897 Sentence denotes The viral genome replication process within the host cell is mainly driven by the NS proteins.
TextSentencer_T38 4803-4897 Sentence denotes The viral genome replication process within the host cell is mainly driven by the NS proteins.
TextSentencer_T39 4898-5045 Sentence denotes 70 The NS1 protein act as an scaffolding protein that anchors the replication complex to the ER 71 membrane and interacts physically with NS4B [4].
TextSentencer_T39 4898-5045 Sentence denotes 70 The NS1 protein act as an scaffolding protein that anchors the replication complex to the ER 71 membrane and interacts physically with NS4B [4].
TextSentencer_T40 5046-5173 Sentence denotes The NS1 protein is a 352-amino-acid 72 polypeptide with a molecular weight of 46-55 kDa, depending on its glycosylation status.
TextSentencer_T40 5046-5173 Sentence denotes The NS1 protein is a 352-amino-acid 72 polypeptide with a molecular weight of 46-55 kDa, depending on its glycosylation status.
TextSentencer_T41 5174-5453 Sentence denotes The 73 NS1 protein exists in multiple oligomeric forms and is found in different cellular locations: a cell 74 membrane-bound form in association with virus-induced intracellular vesicular compartments, 75 on the cell surface and as a soluble secreted hexameric lipoparticle [4].
TextSentencer_T41 5174-5453 Sentence denotes The 73 NS1 protein exists in multiple oligomeric forms and is found in different cellular locations: a cell 74 membrane-bound form in association with virus-induced intracellular vesicular compartments, 75 on the cell surface and as a soluble secreted hexameric lipoparticle [4].
TextSentencer_T42 5454-5599 Sentence denotes The NS1 monomeric 76 form rapidly dimerizes in the endoplasmic reticulum (ER), then three dimeric forms of NS1 77 arrange to form an hexamer [5].
TextSentencer_T42 5454-5599 Sentence denotes The NS1 monomeric 76 form rapidly dimerizes in the endoplasmic reticulum (ER), then three dimeric forms of NS1 77 arrange to form an hexamer [5].
TextSentencer_T43 5600-5751 Sentence denotes The hexameric form of NS1 shows an open barrel form filled 78 with lipids and cholesterol which resemble the lipid composition of the HDL particle [6].
TextSentencer_T43 5600-5751 Sentence denotes The hexameric form of NS1 shows an open barrel form filled 78 with lipids and cholesterol which resemble the lipid composition of the HDL particle [6].
TextSentencer_T44 5752-5926 Sentence denotes 79 Recent studies have shown that the DENV NS1 protein was not only secreted from vertebrate 80 cells, but that was also efficiently secreted from mosquito cells lines [7,8].
TextSentencer_T44 5752-5926 Sentence denotes 79 Recent studies have shown that the DENV NS1 protein was not only secreted from vertebrate 80 cells, but that was also efficiently secreted from mosquito cells lines [7,8].
TextSentencer_T45 5927-6011 Sentence denotes The secretion of NS1 81 in vertebrate cells follows the classical Golgi-pathway [9].
TextSentencer_T45 5927-6011 Sentence denotes The secretion of NS1 81 in vertebrate cells follows the classical Golgi-pathway [9].
TextSentencer_T46 6012-6239 Sentence denotes However, NS1 secretion in infected 82 mosquito cells is associated to a caveolin-1 (CAV-1) dependent pathway and was found to be 83 Brefeldin A (BFA)-insensitive, suggesting a traffic route that bypasses the Golgi-complex [10].
TextSentencer_T46 6012-6239 Sentence denotes However, NS1 secretion in infected 82 mosquito cells is associated to a caveolin-1 (CAV-1) dependent pathway and was found to be 83 Brefeldin A (BFA)-insensitive, suggesting a traffic route that bypasses the Golgi-complex [10].
TextSentencer_T47 6240-6336 Sentence denotes 84 Caveolae are made up of interlocking heteropolymers of a family of small proteins (caveolins:
TextSentencer_T47 6240-6336 Sentence denotes 84 Caveolae are made up of interlocking heteropolymers of a family of small proteins (caveolins:
TextSentencer_T48 6337-6438 Sentence denotes 1-85 3) and a second family of accessory structural proteins (flotillins and three family of cavins).
TextSentencer_T48 6337-6438 Sentence denotes 1-85 3) and a second family of accessory structural proteins (flotillins and three family of cavins).
TextSentencer_T49 6439-6544 Sentence denotes The 86 caveolar architecture is connected with unstructured cavin filaments by coiled-coil domains into .
TextSentencer_T49 6439-6544 Sentence denotes The 86 caveolar architecture is connected with unstructured cavin filaments by coiled-coil domains into .
TextSentencer_T50 6545-6693 Sentence denotes CC-BY 4.0 International license is made available under a The copyright holder for this preprint (which was not peer-reviewed) is the author/funder.
TextSentencer_T50 6545-6693 Sentence denotes CC-BY 4.0 International license is made available under a The copyright holder for this preprint (which was not peer-reviewed) is the author/funder.
TextSentencer_T51 6694-6786 Sentence denotes It . https://doi.org/10.1101/370932 doi: bioRxiv preprint 5 87 a polygonal net-like complex.
TextSentencer_T51 6694-6786 Sentence denotes It . https://doi.org/10.1101/370932 doi: bioRxiv preprint 5 87 a polygonal net-like complex.
TextSentencer_T52 6787-7029 Sentence denotes This complex is believed to provide scaffolding for compartmented 88 cellular processes and participates in multiple cellular functions, including endocytosis, 89 transcytosis, membrane homeostasis, inflammation, and signal transduction [11].
TextSentencer_T52 6787-7029 Sentence denotes This complex is believed to provide scaffolding for compartmented 88 cellular processes and participates in multiple cellular functions, including endocytosis, 89 transcytosis, membrane homeostasis, inflammation, and signal transduction [11].
TextSentencer_T53 7030-7243 Sentence denotes Caveolin 1 90 (Cav-1), a 21-24-kDa scaffolding protein, is not only a key structural component of the caveolae 91 organelle but also plays an important in the transport of free-cholesterol inside the cell [12,13].
TextSentencer_T53 7030-7243 Sentence denotes Caveolin 1 90 (Cav-1), a 21-24-kDa scaffolding protein, is not only a key structural component of the caveolae 91 organelle but also plays an important in the transport of free-cholesterol inside the cell [12,13].
TextSentencer_T54 7244-7432 Sentence denotes 92 The chaperone caveolin complex (CCC) is a cytosolic complex reported to transport cholesterol 93 synthetized de novo from the ER to cell membranes or other compartments within the cell.
TextSentencer_T54 7244-7432 Sentence denotes 92 The chaperone caveolin complex (CCC) is a cytosolic complex reported to transport cholesterol 93 synthetized de novo from the ER to cell membranes or other compartments within the cell.
TextSentencer_T55 7433-7604 Sentence denotes CCC 94 has been described as a complex of CAV-1, Cyclophilin A (CyA), FK506-binding protein 4 or 95 heat shock protein 56 (FKBP52), and Cyclophilin 40 or D (Cy40) [12,14].
TextSentencer_T55 7433-7604 Sentence denotes CCC 94 has been described as a complex of CAV-1, Cyclophilin A (CyA), FK506-binding protein 4 or 95 heat shock protein 56 (FKBP52), and Cyclophilin 40 or D (Cy40) [12,14].
TextSentencer_T56 7605-7719 Sentence denotes Cyclophilin A (CyA), 96 an 18 kDa peptidylprolyl cis-trans isomerase, is a ubiquitous and multifunctional protein.
TextSentencer_T56 7605-7719 Sentence denotes Cyclophilin A (CyA), 96 an 18 kDa peptidylprolyl cis-trans isomerase, is a ubiquitous and multifunctional protein.
TextSentencer_T57 7720-7868 Sentence denotes In 97 addition to its role as a host-cell receptor for cyclosporine A, CyA has diverse functions in 98 inflammatory conditions and diseases [15,16].
TextSentencer_T57 7720-7868 Sentence denotes In 97 addition to its role as a host-cell receptor for cyclosporine A, CyA has diverse functions in 98 inflammatory conditions and diseases [15,16].
TextSentencer_T58 7869-8155 Sentence denotes The 52-kDa FK506-binding protein (FKBP52) an 99 immunophilin belonging to the FKBP family, is a known co-chaperone of the heat shock protein 100 90 (HSP90) and thus may play a role in the intracellular trafficking of hetero-oligomeric forms of 101 the steroid hormone receptors [17,18].
TextSentencer_T58 7869-8155 Sentence denotes The 52-kDa FK506-binding protein (FKBP52) an 99 immunophilin belonging to the FKBP family, is a known co-chaperone of the heat shock protein 100 90 (HSP90) and thus may play a role in the intracellular trafficking of hetero-oligomeric forms of 101 the steroid hormone receptors [17,18].
TextSentencer_T59 8156-8399 Sentence denotes Cyclophilin 40 (Cy40), a member of a family of highly 102 homologous peptidylprolyl cis-trans isomerases (PPIases), is known to play a role in 103 mitochondrial permeability transition (MPT), being an integral constituent of the MPT pore [19].
TextSentencer_T59 8156-8399 Sentence denotes Cyclophilin 40 (Cy40), a member of a family of highly 102 homologous peptidylprolyl cis-trans isomerases (PPIases), is known to play a role in 103 mitochondrial permeability transition (MPT), being an integral constituent of the MPT pore [19].
TextSentencer_T60 8400-8676 Sentence denotes 104 Given the CAV-1 dependent secretion of NS1 protein in mosquito cells and the lipoprotein nature 105 of the released hexameric form of NS1, it was found plausible to study the association of NS1 106 traffic to the novo cholesterol transport in DENV infected mosquito cells.
TextSentencer_T60 8400-8676 Sentence denotes 104 Given the CAV-1 dependent secretion of NS1 protein in mosquito cells and the lipoprotein nature 105 of the released hexameric form of NS1, it was found plausible to study the association of NS1 106 traffic to the novo cholesterol transport in DENV infected mosquito cells.
TextSentencer_T61 8677-8982 Sentence denotes In this work, data is 107 presented indicating that in infected mosquito cells, DENV NS1 enters the unconventional 108 secretory pathway very early after maturation in the ER and usurps the cholesterol transport 109 between the ER and the plasma membrane, mediated by the CCC, to reach the extracellular .
TextSentencer_T61 8677-8982 Sentence denotes In this work, data is 107 presented indicating that in infected mosquito cells, DENV NS1 enters the unconventional 108 secretory pathway very early after maturation in the ER and usurps the cholesterol transport 109 between the ER and the plasma membrane, mediated by the CCC, to reach the extracellular .
TextSentencer_T62 8983-9131 Sentence denotes CC-BY 4.0 International license is made available under a The copyright holder for this preprint (which was not peer-reviewed) is the author/funder.
TextSentencer_T62 8983-9131 Sentence denotes CC-BY 4.0 International license is made available under a The copyright holder for this preprint (which was not peer-reviewed) is the author/funder.
TextSentencer_T63 9132-9202 Sentence denotes It . https://doi.org/10.1101/370932 doi: bioRxiv preprint 6 110 space.
TextSentencer_T63 9132-9202 Sentence denotes It . https://doi.org/10.1101/370932 doi: bioRxiv preprint 6 110 space.
TextSentencer_T64 9203-9351 Sentence denotes In addition, data is presented suggesting that a similar pathway is used for the secretion 111 of Zika Virus NS1 protein in infected mosquito cells.
TextSentencer_T64 9203-9351 Sentence denotes In addition, data is presented suggesting that a similar pathway is used for the secretion 111 of Zika Virus NS1 protein in infected mosquito cells.
TextSentencer_T65 9352-9573 Sentence denotes 112 113 RESULTS 114 NS1 secretion is not affected by drugs that disrupt early steps of the classical secretion 115 pathway 116 Golgicide A (GCA) is a powerful inhibitor of the COPI vehicle transport from ER to Golgi [20].
TextSentencer_T65 9352-9573 Sentence denotes 112 113 RESULTS 114 NS1 secretion is not affected by drugs that disrupt early steps of the classical secretion 115 pathway 116 Golgicide A (GCA) is a powerful inhibitor of the COPI vehicle transport from ER to Golgi [20].
TextSentencer_T66 9574-9841 Sentence denotes 117 Thus, the cytotoxicity of GCA in the mosquito cell lines (C6/36, Aag2) and the vertebrate cell line 118 BHK-21, used for comparisons, was measured using the reduction of tetrazolium salts to 119 examine proliferation in cells treated with serial dilutions of GCA.
TextSentencer_T66 9574-9841 Sentence denotes 117 Thus, the cytotoxicity of GCA in the mosquito cell lines (C6/36, Aag2) and the vertebrate cell line 118 BHK-21, used for comparisons, was measured using the reduction of tetrazolium salts to 119 examine proliferation in cells treated with serial dilutions of GCA.
TextSentencer_T67 9842-9947 Sentence denotes No significant cytotoxicity was 120 observed under 30 M of GCA in any of the three cell types (Fig 1A) .
TextSentencer_T67 9842-9947 Sentence denotes No significant cytotoxicity was 120 observed under 30 M of GCA in any of the three cell types (Fig 1A) .
TextSentencer_T68 9948-10060 Sentence denotes Fli-06 is a novel drug which 121 inhibits the diffusion of ER synthetized proteins to the ER-exit sites [21,22].
TextSentencer_T68 9948-10060 Sentence denotes Fli-06 is a novel drug which 121 inhibits the diffusion of ER synthetized proteins to the ER-exit sites [21,22].
TextSentencer_T69 10061-10167 Sentence denotes Fli-06 cytotoxicity 122 was determined in C6/36 and Aag2 cells also using also tetrazolium salt reduction.
TextSentencer_T69 10061-10167 Sentence denotes Fli-06 cytotoxicity 122 was determined in C6/36 and Aag2 cells also using also tetrazolium salt reduction.
TextSentencer_T70 10168-10274 Sentence denotes No significant 123 cytotoxicity was observed under 100 M of Fli-06 in the mosquito cells lines (Fig 1B) .
TextSentencer_T70 10168-10274 Sentence denotes No significant 123 cytotoxicity was observed under 100 M of Fli-06 in the mosquito cells lines (Fig 1B) .
TextSentencer_T71 10275-10464 Sentence denotes Thus, 124 concentration of 27 M and 100 M were used for GCA and Fli-06, respectively, since these 125 concentrations proved non-toxic, yet effective in causing Golgi disruption (Fig 1C) .
TextSentencer_T71 10275-10464 Sentence denotes Thus, 124 concentration of 27 M and 100 M were used for GCA and Fli-06, respectively, since these 125 concentrations proved non-toxic, yet effective in causing Golgi disruption (Fig 1C) .
TextSentencer_T72 10465-10601 Sentence denotes Figures 1D 126 and 1E show that the GCA-treatment of DENV infected C6/36 and Aag2 cells did not cause any 127 change in NS1 secretion.
TextSentencer_T72 10465-10601 Sentence denotes Figures 1D 126 and 1E show that the GCA-treatment of DENV infected C6/36 and Aag2 cells did not cause any 127 change in NS1 secretion.
TextSentencer_T73 10602-10720 Sentence denotes In contrast, NS1 secretion was reduced over 70% in infected BHK-21 128 cell treated either with GCA or BFA ( Fig 1F) .
TextSentencer_T73 10602-10720 Sentence denotes In contrast, NS1 secretion was reduced over 70% in infected BHK-21 128 cell treated either with GCA or BFA ( Fig 1F) .
TextSentencer_T74 10721-10846 Sentence denotes As expected, virion release was reduced in 129 mosquito and vertebrate cells after GCA or BFA treatment (Fig 1G, 1H and 1I ).
TextSentencer_T74 10721-10846 Sentence denotes As expected, virion release was reduced in 129 mosquito and vertebrate cells after GCA or BFA treatment (Fig 1G, 1H and 1I ).
TextSentencer_T75 10847-10964 Sentence denotes To further explore 130 the traffic route of NS1 in DENV infected mosquito cells, cells were also treated with Fli-06.
TextSentencer_T75 10847-10964 Sentence denotes To further explore 130 the traffic route of NS1 in DENV infected mosquito cells, cells were also treated with Fli-06.
TextSentencer_T76 10965-11068 Sentence denotes 131 Treatment of infected C6/36 or Aag2 cells with non-toxic concentrations of Fli-06 shows no effect .
TextSentencer_T76 10965-11068 Sentence denotes 131 Treatment of infected C6/36 or Aag2 cells with non-toxic concentrations of Fli-06 shows no effect .
TextSentencer_T77 11069-11217 Sentence denotes CC-BY 4.0 International license is made available under a The copyright holder for this preprint (which was not peer-reviewed) is the author/funder.
TextSentencer_T77 11069-11217 Sentence denotes CC-BY 4.0 International license is made available under a The copyright holder for this preprint (which was not peer-reviewed) is the author/funder.
TextSentencer_T78 11218-11359 Sentence denotes It . https://doi.org/10.1101/370932 doi: bioRxiv preprint 7 132 in NS1 secretion, while significantly affecting virion release (Fig 1G, 1H) .
TextSentencer_T78 11218-11359 Sentence denotes It . https://doi.org/10.1101/370932 doi: bioRxiv preprint 7 132 in NS1 secretion, while significantly affecting virion release (Fig 1G, 1H) .
TextSentencer_T79 11360-11445 Sentence denotes Experiments with Fli-133 06 were not carried out in BHK-21 due to high drug toxicity.
TextSentencer_T79 11360-11445 Sentence denotes Experiments with Fli-133 06 were not carried out in BHK-21 due to high drug toxicity.
TextSentencer_T80 11446-11672 Sentence denotes The results with GCA and Fli-06 134 indicate NS1 is not secreted by a classical secretory pathway and that newly synthetized DENV 135 NS1 do not reach the ER-exit sites and leaves the ER compartment very early after synthesis.
TextSentencer_T80 11446-11672 Sentence denotes The results with GCA and Fli-06 134 indicate NS1 is not secreted by a classical secretory pathway and that newly synthetized DENV 135 NS1 do not reach the ER-exit sites and leaves the ER compartment very early after synthesis.
TextSentencer_T81 11673-11915 Sentence denotes 136 Since the release of mature virions, which are multimolecular complexes, may not be equivalent 137 to the release of a single protein, an additional control for protein trafficking through the classical 138 secretory pathway was included.
TextSentencer_T81 11673-11915 Sentence denotes 136 Since the release of mature virions, which are multimolecular complexes, may not be equivalent 137 to the release of a single protein, an additional control for protein trafficking through the classical 138 secretory pathway was included.
TextSentencer_T82 11916-12050 Sentence denotes To this aim, a vector, pAc5-mCherry-GLuc-neo, constitutively 139 expressing Gaussia Luciferase in C6/36 cell was constructed (S1 Fig).
TextSentencer_T82 11916-12050 Sentence denotes To this aim, a vector, pAc5-mCherry-GLuc-neo, constitutively 139 expressing Gaussia Luciferase in C6/36 cell was constructed (S1 Fig).
TextSentencer_T83 12051-12249 Sentence denotes To evaluate the effect of 140 GCA treatment on GLuc secretion, cells were treated with GCA for 24 h and the activity of GLuc 141 present in the cell supernatants analyzed using a luminescence assay.
TextSentencer_T83 12051-12249 Sentence denotes To evaluate the effect of 140 GCA treatment on GLuc secretion, cells were treated with GCA for 24 h and the activity of GLuc 141 present in the cell supernatants analyzed using a luminescence assay.
TextSentencer_T84 12250-12465 Sentence denotes A decrease in luciferase 142 activity, in relation to the control (DMSO), was detected upon treatment of C6/36 with GCA, and 143 in BHK-21 cells transfected with ptk-GLuc vector, run in parallel as control (Fig.1J).
TextSentencer_T84 12250-12465 Sentence denotes A decrease in luciferase 142 activity, in relation to the control (DMSO), was detected upon treatment of C6/36 with GCA, and 143 in BHK-21 cells transfected with ptk-GLuc vector, run in parallel as control (Fig.1J).
TextSentencer_T85 12466-12770 Sentence denotes These results 144 indicated that the luciferase reporter is secreted in both mosquito and vertebrate cells following 145 a classical secretory route and reinforce the results indicating that in mosquito cells DENV NS1 146 is secreted by an unconventional secretory route that bypasses the Golgi (Fig 1K).
TextSentencer_T85 12466-12770 Sentence denotes These results 144 indicated that the luciferase reporter is secreted in both mosquito and vertebrate cells following 145 a classical secretory route and reinforce the results indicating that in mosquito cells DENV NS1 146 is secreted by an unconventional secretory route that bypasses the Golgi (Fig 1K).
TextSentencer_T86 12771-12917 Sentence denotes 147 148 NS1 secretion is dependent on the CCC 149 The CCC is formed by the association of CAV-1 with the chaperones CyA, FKBP52, Cy40 150 [12,13].
TextSentencer_T86 12771-12917 Sentence denotes 147 148 NS1 secretion is dependent on the CCC 149 The CCC is formed by the association of CAV-1 with the chaperones CyA, FKBP52, Cy40 150 [12,13].
TextSentencer_T87 12918-12983 Sentence denotes The CCC is responsible for cholesterol transport inside the cell.
TextSentencer_T87 12918-12983 Sentence denotes The CCC is responsible for cholesterol transport inside the cell.
TextSentencer_T88 12984-13065 Sentence denotes Previous data by us [10] 151 indicated that NS1 secretion was dependent on CAV-1.
TextSentencer_T88 12984-13065 Sentence denotes Previous data by us [10] 151 indicated that NS1 secretion was dependent on CAV-1.
TextSentencer_T89 13066-13298 Sentence denotes Thus, to fully evaluate the participation 152 of the CCC in DENV NS1 secretion from mosquito cells, CyA was pharmacologically inhibited 153 by treatment with Cyclosporin A (CsA) and the expression of the chaperones FKBP52 and Cy40 .
TextSentencer_T89 13066-13298 Sentence denotes Thus, to fully evaluate the participation 152 of the CCC in DENV NS1 secretion from mosquito cells, CyA was pharmacologically inhibited 153 by treatment with Cyclosporin A (CsA) and the expression of the chaperones FKBP52 and Cy40 .
TextSentencer_T90 13299-13356 Sentence denotes CC-BY 4.0 International license is made available under a
TextSentencer_T90 13299-13356 Sentence denotes CC-BY 4.0 International license is made available under a
TextSentencer_T91 13357-13447 Sentence denotes The copyright holder for this preprint (which was not peer-reviewed) is the author/funder.
TextSentencer_T91 13357-13447 Sentence denotes The copyright holder for this preprint (which was not peer-reviewed) is the author/funder.
TextSentencer_T92 13448-13538 Sentence denotes It . https://doi.org/10.1101/370932 doi: bioRxiv preprint 8 154 knocked down using siRNAs.
TextSentencer_T92 13448-13538 Sentence denotes It . https://doi.org/10.1101/370932 doi: bioRxiv preprint 8 154 knocked down using siRNAs.
TextSentencer_T93 13539-13654 Sentence denotes Previously, the non-cytotoxic concentration for CsA in mosquito 155 and vertebrate cells were determined (Fig 2A) .
TextSentencer_T93 13539-13654 Sentence denotes Previously, the non-cytotoxic concentration for CsA in mosquito 155 and vertebrate cells were determined (Fig 2A) .
TextSentencer_T94 13655-13847 Sentence denotes DENV2 or DENV4-infected C6/36 or Aag2 cells 156 were treated with 9 M CsA for 24 hours; NS1 secretion was reduced in about 30% and 40% in 157 C6/36 and Aag2 cells, respectively (Fig 2B, 2E) .
TextSentencer_T94 13655-13847 Sentence denotes DENV2 or DENV4-infected C6/36 or Aag2 cells 156 were treated with 9 M CsA for 24 hours; NS1 secretion was reduced in about 30% and 40% in 157 C6/36 and Aag2 cells, respectively (Fig 2B, 2E) .
TextSentencer_T95 13848-13954 Sentence denotes However, virion release in the treated mosquito 158 cells was not significantly affected (Fig 2C and 2F ).
TextSentencer_T95 13848-13954 Sentence denotes However, virion release in the treated mosquito 158 cells was not significantly affected (Fig 2C and 2F ).
TextSentencer_T96 13955-14085 Sentence denotes In contrast, the inhibition of CyA in BHK-21 159 cells did not have any effect in either NS1 or virion secretion (Figs 2H and 2I).
TextSentencer_T96 13955-14085 Sentence denotes In contrast, the inhibition of CyA in BHK-21 159 cells did not have any effect in either NS1 or virion secretion (Figs 2H and 2I).
TextSentencer_T97 14086-14253 Sentence denotes To further 160 corroborate the participation of CyA in cholesterol and lipid traffic, the lipid droplet count in CsA 161 treated versus untreated cells was determined.
TextSentencer_T97 14086-14253 Sentence denotes To further 160 corroborate the participation of CyA in cholesterol and lipid traffic, the lipid droplet count in CsA 161 treated versus untreated cells was determined.
TextSentencer_T98 14254-14430 Sentence denotes In all cases, C6/36, Aag2 and BHK-21 cells 162 treated with CsA showed a diminished amount of lipid droplets per cell as compared with un-163 treated cells (Fig 2D, 2G and 2J).
TextSentencer_T98 14254-14430 Sentence denotes In all cases, C6/36, Aag2 and BHK-21 cells 162 treated with CsA showed a diminished amount of lipid droplets per cell as compared with un-163 treated cells (Fig 2D, 2G and 2J).
TextSentencer_T99 14431-14512 Sentence denotes 164 The expression of FKBP52 in C6/36 cells was knock down by siRNA transfection.
TextSentencer_T99 14431-14512 Sentence denotes 164 The expression of FKBP52 in C6/36 cells was knock down by siRNA transfection.
TextSentencer_T100 14513-14592 Sentence denotes Optimal gene 165 knock down was achieved after 48h post transfection (Fig 3A) .
TextSentencer_T100 14513-14592 Sentence denotes Optimal gene 165 knock down was achieved after 48h post transfection (Fig 3A) .
TextSentencer_T101 14593-14845 Sentence denotes The effect of FKPB52 reduced 166 expression in cholesterol and lipid metabolism was evaluated measuring the lipid droplet 167 counting per cell and a reduction of about 50% in the number of lipid droplets was observed after 168 the treatment (Fig 3B) .
TextSentencer_T101 14593-14845 Sentence denotes The effect of FKPB52 reduced 166 expression in cholesterol and lipid metabolism was evaluated measuring the lipid droplet 167 counting per cell and a reduction of about 50% in the number of lipid droplets was observed after 168 the treatment (Fig 3B) .
TextSentencer_T102 14846-14964 Sentence denotes In cells treated with siRNA FKBP52, a significant reduction in NS1 of 169 about 20% secretion was observed ( Fig 3C) .
TextSentencer_T102 14846-14964 Sentence denotes In cells treated with siRNA FKBP52, a significant reduction in NS1 of 169 about 20% secretion was observed ( Fig 3C) .
TextSentencer_T103 14965-15031 Sentence denotes Viral release was not affected by FKBP52-knock 170 down (Fig 3D) .
TextSentencer_T103 14965-15031 Sentence denotes Viral release was not affected by FKBP52-knock 170 down (Fig 3D) .
TextSentencer_T104 15032-15212 Sentence denotes Given that FKBP52 is a co-chaperone of HSP90 [17,23], we wanted to discard if 171 the observed inhibitory effect on NS1 secretion might be related to a reduction in HSP90 activity.
TextSentencer_T104 15032-15212 Sentence denotes Given that FKBP52 is a co-chaperone of HSP90 [17,23], we wanted to discard if 171 the observed inhibitory effect on NS1 secretion might be related to a reduction in HSP90 activity.
TextSentencer_T105 15213-15336 Sentence denotes 172 Thus, the effect of the HSP90 inhibitor geldanamycin (GA) in NS1 release from DENV infected 173 C6/36 cells was tested.
TextSentencer_T105 15213-15336 Sentence denotes 172 Thus, the effect of the HSP90 inhibitor geldanamycin (GA) in NS1 release from DENV infected 173 C6/36 cells was tested.
TextSentencer_T106 15337-15415 Sentence denotes As shown in Figure 3E , NS1 secretion was not affected after GA 174 treatment.
TextSentencer_T106 15337-15415 Sentence denotes As shown in Figure 3E , NS1 secretion was not affected after GA 174 treatment.
TextSentencer_T107 15416-15534 Sentence denotes The expected reduction in virus yield observed in GA treated cell (Fig 3F) show that 175 effective GA doses were used.
TextSentencer_T107 15416-15534 Sentence denotes The expected reduction in virus yield observed in GA treated cell (Fig 3F) show that 175 effective GA doses were used.
TextSentencer_T108 15535-15696 Sentence denotes These results indicate that the effect in NS1 secretion observed 176 after FKBP52 silencing is not related with HSP90 activity and was rather associated with the
TextSentencer_T108 15535-15696 Sentence denotes These results indicate that the effect in NS1 secretion observed 176 after FKBP52 silencing is not related with HSP90 activity and was rather associated with the
TextSentencer_T109 15698-15788 Sentence denotes The copyright holder for this preprint (which was not peer-reviewed) is the author/funder.
TextSentencer_T109 15698-15788 Sentence denotes The copyright holder for this preprint (which was not peer-reviewed) is the author/funder.
TextSentencer_T110 15789-15873 Sentence denotes It . https://doi.org/10.1101/370932 doi: bioRxiv preprint 9 177 function of the CCC.
TextSentencer_T110 15789-15873 Sentence denotes It . https://doi.org/10.1101/370932 doi: bioRxiv preprint 9 177 function of the CCC.
TextSentencer_T111 15874-15980 Sentence denotes Finally, the expression of Cy40 was significantly knock down at 48h post 178 siRNA transfection (Fig 4A) .
TextSentencer_T111 15874-15980 Sentence denotes Finally, the expression of Cy40 was significantly knock down at 48h post 178 siRNA transfection (Fig 4A) .
TextSentencer_T112 15981-16091 Sentence denotes A reduction in about 60% of lipid droplets counting per cell was 179 observed in transfected cells ( Fig 4B) .
TextSentencer_T112 15981-16091 Sentence denotes A reduction in about 60% of lipid droplets counting per cell was 179 observed in transfected cells ( Fig 4B) .
TextSentencer_T113 16092-16189 Sentence denotes Yet, neither NS1 secretion nor virus release was affected 180 by Cy40 silencing (Fig 4C and 4D ).
TextSentencer_T113 16092-16189 Sentence denotes Yet, neither NS1 secretion nor virus release was affected 180 by Cy40 silencing (Fig 4C and 4D ).
TextSentencer_T114 16190-16307 Sentence denotes This result suggests that while Cy40 participates in 181 cholesterol traffic it may be dispensable for NS1 secretion.
TextSentencer_T114 16190-16307 Sentence denotes This result suggests that while Cy40 participates in 181 cholesterol traffic it may be dispensable for NS1 secretion.
TextSentencer_T115 16308-16398 Sentence denotes The copyright holder for this preprint (which was not peer-reviewed) is the author/funder.
TextSentencer_T115 16308-16398 Sentence denotes The copyright holder for this preprint (which was not peer-reviewed) is the author/funder.
TextSentencer_T116 16399-16529 Sentence denotes It . https://doi.org/10.1101/370932 doi: bioRxiv preprint 13 266 were detected in BHK-21 cells with ZIKV and YFV (data not shown).
TextSentencer_T116 16399-16529 Sentence denotes It . https://doi.org/10.1101/370932 doi: bioRxiv preprint 13 266 were detected in BHK-21 cells with ZIKV and YFV (data not shown).
TextSentencer_T117 16530-16651 Sentence denotes The secretion of YFV NS1 267 was found to be sensitive to BFA treatment in both mosquito and vertebrate cells (Fig. 9A) .
TextSentencer_T117 16530-16651 Sentence denotes The secretion of YFV NS1 267 was found to be sensitive to BFA treatment in both mosquito and vertebrate cells (Fig. 9A) .
TextSentencer_T118 16652-16821 Sentence denotes In 268 contrast, the secretion of ZIKV NS1 was found to be insensitive to BFA and GCA treatment in 269 infected mosquito, but not in infected vertebrate cells (Fig 9B) .
TextSentencer_T118 16652-16821 Sentence denotes In 268 contrast, the secretion of ZIKV NS1 was found to be insensitive to BFA and GCA treatment in 269 infected mosquito, but not in infected vertebrate cells (Fig 9B) .
TextSentencer_T119 16822-16970 Sentence denotes In addition, the secretion of ZIKV 270 NS1 was reduced in mosquito, but not in vertebrate cells, treated with the CyA inhibitor CsA (Fig 271 9C ).
TextSentencer_T119 16822-16970 Sentence denotes In addition, the secretion of ZIKV 270 NS1 was reduced in mosquito, but not in vertebrate cells, treated with the CyA inhibitor CsA (Fig 271 9C ).
TextSentencer_T120 16971-17234 Sentence denotes Co-localizations experiments carried out in C6/36, Aag2 and Vero-E6 cells infected with 272 ZIKV and YFV, showed a stronger, and a significantly different, co-localization between ZIKV 273 NS1 and CAV-1 than between YFV NS1 and CAV-1, in both mosquito cell lines.
TextSentencer_T120 16971-17234 Sentence denotes Co-localizations experiments carried out in C6/36, Aag2 and Vero-E6 cells infected with 272 ZIKV and YFV, showed a stronger, and a significantly different, co-localization between ZIKV 273 NS1 and CAV-1 than between YFV NS1 and CAV-1, in both mosquito cell lines.
TextSentencer_T121 17235-17346 Sentence denotes No co-274 localization of ZIKV or YFV NS1 with CAV-1 was observed in vertebrate cell line ( Fig 9D and 275 9E).
TextSentencer_T121 17235-17346 Sentence denotes No co-274 localization of ZIKV or YFV NS1 with CAV-1 was observed in vertebrate cell line ( Fig 9D and 275 9E).
TextSentencer_T122 17347-17528 Sentence denotes Of note, the co-localization data support the predictions derived from the presence of a 276 conserved caveolin binding (CBD) domain in ZIKV (and DENV) but not in YFV NS1 (Fig 9F) .
TextSentencer_T122 17347-17528 Sentence denotes Of note, the co-localization data support the predictions derived from the presence of a 276 conserved caveolin binding (CBD) domain in ZIKV (and DENV) but not in YFV NS1 (Fig 9F) .
TextSentencer_T123 17529-17619 Sentence denotes The copyright holder for this preprint (which was not peer-reviewed) is the author/funder.
TextSentencer_T123 17529-17619 Sentence denotes The copyright holder for this preprint (which was not peer-reviewed) is the author/funder.
TextSentencer_T124 17620-17730 Sentence denotes It . https://doi.org/10.1101/370932 doi: bioRxiv preprint 14 289 10.2210/pdb2a65/pdb) with a p-value 2.69e-03.
TextSentencer_T124 17620-17730 Sentence denotes It . https://doi.org/10.1101/370932 doi: bioRxiv preprint 14 289 10.2210/pdb2a65/pdb) with a p-value 2.69e-03.
TextSentencer_T125 17731-17979 Sentence denotes Molecular docking predicts a favorable 290 interaction between the NS1 hydrophobic domain including the grease fingers, where the CBD 291 is located, with a pocket in the CAV-1 scaffolding domain (CSD) with distances less 8Ǻ ( Fig 9A 292 and 9B).
TextSentencer_T125 17731-17979 Sentence denotes Molecular docking predicts a favorable 290 interaction between the NS1 hydrophobic domain including the grease fingers, where the CBD 291 is located, with a pocket in the CAV-1 scaffolding domain (CSD) with distances less 8Ǻ ( Fig 9A 292 and 9B).
TextSentencer_T126 17980-18186 Sentence denotes The molecular docking results obtained with CAV-1 and NS1 were used as a control to 293 carry on additional docking experiments between the DENV NS1 or CAV-1 and the CCC 294 chaperones FKBP52, Cy40 and CyA.
TextSentencer_T126 17980-18186 Sentence denotes The molecular docking results obtained with CAV-1 and NS1 were used as a control to 293 carry on additional docking experiments between the DENV NS1 or CAV-1 and the CCC 294 chaperones FKBP52, Cy40 and CyA.
TextSentencer_T127 18187-18399 Sentence denotes Calculations of the energy binding of several docking 295 clusters showed that the DENV NS1 has the lowest and more favorable energy for CAV-1, 296 compared to the energy binding obtained for FKBP52, Cy40 or CyA.
TextSentencer_T127 18187-18399 Sentence denotes Calculations of the energy binding of several docking 295 clusters showed that the DENV NS1 has the lowest and more favorable energy for CAV-1, 296 compared to the energy binding obtained for FKBP52, Cy40 or CyA.
TextSentencer_T128 18400-18579 Sentence denotes In addition, lower energies 297 were obtained for the binding of CAV-1 with the other chaperones of the CCC, than for the 298 binding of NS1 with those same chaperones ( Fig 9C) .
TextSentencer_T128 18400-18579 Sentence denotes In addition, lower energies 297 were obtained for the binding of CAV-1 with the other chaperones of the CCC, than for the 298 binding of NS1 with those same chaperones ( Fig 9C) .
TextSentencer_T129 18580-18819 Sentence denotes Based in the free energies data, we predict 299 a model of interaction of NS1 with the CCC proteins in which FKBP52, Cy40 and CyA are directly 300 bound to CAV-1, and NS1 is bound to the CCC through the CAV-1 scaffolding domain ( Fig 9D) .
TextSentencer_T129 18580-18819 Sentence denotes Based in the free energies data, we predict 299 a model of interaction of NS1 with the CCC proteins in which FKBP52, Cy40 and CyA are directly 300 bound to CAV-1, and NS1 is bound to the CCC through the CAV-1 scaffolding domain ( Fig 9D) .
TextSentencer_T130 18820-18910 Sentence denotes The copyright holder for this preprint (which was not peer-reviewed) is the author/funder.
TextSentencer_T130 18820-18910 Sentence denotes The copyright holder for this preprint (which was not peer-reviewed) is the author/funder.
TextSentencer_T131 18911-19207 Sentence denotes It . https://doi.org/10.1101/370932 doi: bioRxiv preprint 15 303 DISCUSSION 304 Our previous work demonstrated that DENV NS1 secretion in mosquito cell lines follows a BFA-305 insensitivity secretion pathway that depends on CAV-1; moreover, NS1 was found to interact 306 directly with CAV-1 [10].
TextSentencer_T131 18911-19207 Sentence denotes It . https://doi.org/10.1101/370932 doi: bioRxiv preprint 15 303 DISCUSSION 304 Our previous work demonstrated that DENV NS1 secretion in mosquito cell lines follows a BFA-305 insensitivity secretion pathway that depends on CAV-1; moreover, NS1 was found to interact 306 directly with CAV-1 [10].
TextSentencer_T132 19208-19430 Sentence denotes Therefore, in the present work, we explored the association of NS1 with 307 proteins involved, together with CAV-1, in the intracellular transport of cholesterol and the 308 dependence of NS1 secretion on such association.
TextSentencer_T132 19208-19430 Sentence denotes Therefore, in the present work, we explored the association of NS1 with 307 proteins involved, together with CAV-1, in the intracellular transport of cholesterol and the 308 dependence of NS1 secretion on such association.
TextSentencer_T133 19431-19651 Sentence denotes The results revealed that NS1 is associated 309 to the chaperone caveolin complex (CAV-1, Cy40, FKBP52 and CyA) and that the integrity of 310 this complex is necessary for the secretion of NS1 in mosquito infected cells.
TextSentencer_T133 19431-19651 Sentence denotes The results revealed that NS1 is associated 309 to the chaperone caveolin complex (CAV-1, Cy40, FKBP52 and CyA) and that the integrity of 310 this complex is necessary for the secretion of NS1 in mosquito infected cells.
TextSentencer_T134 19652-19850 Sentence denotes Additional data 311 suggest that in virus infected mosquito cells, ZIKV NS1, but not yellow fever virus NS1, also 312 associates with CAV-1 and usurps the cholesterol traffic pathways for secretion.
TextSentencer_T134 19652-19850 Sentence denotes Additional data 311 suggest that in virus infected mosquito cells, ZIKV NS1, but not yellow fever virus NS1, also 312 associates with CAV-1 and usurps the cholesterol traffic pathways for secretion.
TextSentencer_T135 19851-20055 Sentence denotes Thus, the lipid 313 content or molecular structure of the DENV NS1 seems to have taken advantage of mosquito 314 cell cholesterol transport to guarantee the efficient NS1 secretion in the mosquito vector.
TextSentencer_T135 19851-20055 Sentence denotes Thus, the lipid 313 content or molecular structure of the DENV NS1 seems to have taken advantage of mosquito 314 cell cholesterol transport to guarantee the efficient NS1 secretion in the mosquito vector.
TextSentencer_T136 20056-20146 Sentence denotes The copyright holder for this preprint (which was not peer-reviewed) is the author/funder.
TextSentencer_T136 20056-20146 Sentence denotes The copyright holder for this preprint (which was not peer-reviewed) is the author/funder.
TextSentencer_T137 20147-20293 Sentence denotes It . https://doi.org/10.1101/370932 doi: bioRxiv preprint 16 325 Few works have reported a luciferase secretion system in insect cell lines [26] .
TextSentencer_T137 20147-20293 Sentence denotes It . https://doi.org/10.1101/370932 doi: bioRxiv preprint 16 325 Few works have reported a luciferase secretion system in insect cell lines [26] .
TextSentencer_T138 20294-20537 Sentence denotes Yet, as an 326 additional tool to study classical secretion in mosquito cell lines, an efficient Gaussia luciferase 327 reporter system that enables the detection of Luciferase in cell supernatants 6 hours after 328 transfection was developed.
TextSentencer_T138 20294-20537 Sentence denotes Yet, as an 326 additional tool to study classical secretion in mosquito cell lines, an efficient Gaussia luciferase 327 reporter system that enables the detection of Luciferase in cell supernatants 6 hours after 328 transfection was developed.
TextSentencer_T139 20538-20811 Sentence denotes The secretion of luciferase was significantly affected after treatment 329 with GCA and BFA (data not shown); these data, together with the observed inhibition of viral 330 particle release, indicates the existence of a robust classical secretion pathway in mosquito cells.
TextSentencer_T139 20538-20811 Sentence denotes The secretion of luciferase was significantly affected after treatment 329 with GCA and BFA (data not shown); these data, together with the observed inhibition of viral 330 particle release, indicates the existence of a robust classical secretion pathway in mosquito cells.
TextSentencer_T140 20812-20902 Sentence denotes The copyright holder for this preprint (which was not peer-reviewed) is the author/funder.
TextSentencer_T140 20812-20902 Sentence denotes The copyright holder for this preprint (which was not peer-reviewed) is the author/funder.
TextSentencer_T141 20903-21010 Sentence denotes It . https://doi.org/10.1101/370932 doi: bioRxiv preprint 18 371 (CSD) with a distance of 5Ǻ approximately.
TextSentencer_T141 20903-21010 Sentence denotes It . https://doi.org/10.1101/370932 doi: bioRxiv preprint 18 371 (CSD) with a distance of 5Ǻ approximately.
TextSentencer_T142 21011-21178 Sentence denotes Further analysis also showed, high energy binding 372 through hydrophobic dynamics, between the NS1 -roll domain and FKBP52, CyA and Cy40 373 with distances under 6Ǻ.
TextSentencer_T142 21011-21178 Sentence denotes Further analysis also showed, high energy binding 372 through hydrophobic dynamics, between the NS1 -roll domain and FKBP52, CyA and Cy40 373 with distances under 6Ǻ.
TextSentencer_T143 21179-21330 Sentence denotes However, the model predicts all these proteins interacting at the same 374 domain in NS1 (data not shown), which seems unlikely, due to steric effects.
TextSentencer_T143 21179-21330 Sentence denotes However, the model predicts all these proteins interacting at the same 374 domain in NS1 (data not shown), which seems unlikely, due to steric effects.
TextSentencer_T144 21331-21530 Sentence denotes Thus, we favor an 375 alternative molecular docking model, based on interaction energies, in which NS1 is bound 376 directly to CAV-1, and in turn FKBP52, CyA and Cy40 are bound to CAV-1 (Fig. 10D ).
TextSentencer_T144 21331-21530 Sentence denotes Thus, we favor an 375 alternative molecular docking model, based on interaction energies, in which NS1 is bound 376 directly to CAV-1, and in turn FKBP52, CyA and Cy40 are bound to CAV-1 (Fig. 10D ).
TextSentencer_T145 21531-21610 Sentence denotes This 377 model still allowed interaction of NS1 with the CCC through membranes.
TextSentencer_T145 21531-21610 Sentence denotes This 377 model still allowed interaction of NS1 with the CCC through membranes.
TextSentencer_T146 21611-21757 Sentence denotes However, if the NS1-378 CCC complex will traffic to the plasma membrane as a free-cytosolic complex or membrane 379 associated is totally unknown.
TextSentencer_T146 21611-21757 Sentence denotes However, if the NS1-378 CCC complex will traffic to the plasma membrane as a free-cytosolic complex or membrane 379 associated is totally unknown.
TextSentencer_T147 21758-21848 Sentence denotes The copyright holder for this preprint (which was not peer-reviewed) is the author/funder.
TextSentencer_T147 21758-21848 Sentence denotes The copyright holder for this preprint (which was not peer-reviewed) is the author/funder.
TextSentencer_T148 21849-21908 Sentence denotes It . https://doi.org/10.1101/370932 doi: bioRxiv preprint .
TextSentencer_T148 21849-21908 Sentence denotes It . https://doi.org/10.1101/370932 doi: bioRxiv preprint .
TextSentencer_T149 21909-22057 Sentence denotes CC-BY 4.0 International license is made available under a The copyright holder for this preprint (which was not peer-reviewed) is the author/funder.
TextSentencer_T149 21909-22057 Sentence denotes CC-BY 4.0 International license is made available under a The copyright holder for this preprint (which was not peer-reviewed) is the author/funder.
TextSentencer_T150 22058-22148 Sentence denotes The copyright holder for this preprint (which was not peer-reviewed) is the author/funder.
TextSentencer_T150 22058-22148 Sentence denotes The copyright holder for this preprint (which was not peer-reviewed) is the author/funder.
TextSentencer_T151 22149-22206 Sentence denotes It . https://doi.org/10.1101/370932 doi: bioRxiv preprint
TextSentencer_T151 22149-22206 Sentence denotes It . https://doi.org/10.1101/370932 doi: bioRxiv preprint