PMC:7464876 / 38426-39751 JSONTXT

{"target":"https://pubannotation.org/docs/sourcedb/PMC/sourceid/7464876","sourcedb":"PMC","sourceid":"7464876","source_url":"https://www.ncbi.nlm.nih.gov/pmc/7464876","text":"3.7. HBV L Interacts with ERGIC-53 in a N146-Glycan-Dependent Manner\nAccordingly, we next investigated whether there might be physical links between the viral N-glycan and cellular ERGIC-53. As above, HuH-7 cells were depleted for Sec24A in order to arrest ERGIC-53 within the ER, following a retransfection with the wt replicon construct or the envelope-defective HBV.Envminus replicon trans-complemented with Env.N146D. In control cells, neither the wt L nor the L.N146D mutant exhibited any colocalization with the tubulovesicular clusters stained with anti-ERGIC-53 (Figure 7). Upon depletion of Sec24A, the ERGIC-53-specific labeling pattern changed and again appeared in ER-like reticular areas. Importantly, this pattern strongly coincided with the CS structure of wt L (Figure 7A) but poorly with the CS structure of L.N146D (Figure 7B). To corroborate these findings, CO-IP analyses were performed. Since ERGIC-53/cargo interactions are known to be transient and short-lived [42,50], we here used the ectopic expression of a Myc-tagged ERGIC-53 construct together with the wt or mutant HBV replicons. As shown in Figure 7C, wt L, but not L.N146D, efficiently coprecipitated ERGIC-53. Together, these results point to a productive interaction between L and ERGIC-53 that is primary mediated by the N146-linked glycan.","divisions":[{"label":"title","span":{"begin":0,"end":68}}],"tracks":[{"project":"2_test","denotations":[{"id":"32806600-10559958-20296233","span":{"begin":985,"end":987},"obj":"10559958"},{"id":"32806600-19609866-20296234","span":{"begin":988,"end":990},"obj":"19609866"}],"attributes":[{"subj":"32806600-10559958-20296233","pred":"source","obj":"2_test"},{"subj":"32806600-19609866-20296234","pred":"source","obj":"2_test"}]}],"config":{"attribute types":[{"pred":"source","value type":"selection","values":[{"id":"2_test","color":"#ec93d5","default":true}]}]}}