Id |
Subject |
Object |
Predicate |
Lexical cue |
T120 |
0-145 |
Sentence |
denotes |
The presence of multiple intrinsically disordered regions in the N protein precluded the determination of its structure by X-ray crystallography. |
T121 |
146-295 |
Sentence |
denotes |
Instead, we used rigid body modeling of the SAXS data with the N-terminal domain (NTD; solved in this study) and the C-terminal domains (CTD, PDB ID: |
T122 |
296-409 |
Sentence |
denotes |
6G13).23 In this way, we obtained structural models for the free N protein and its complex with P3 (Figure 3B,C). |
T123 |
410-439 |
Sentence |
denotes |
Excellent fits were obtained. |
T124 |
440-560 |
Sentence |
denotes |
Representative structural models for the full-length protein without and with P3 are shown in Figure 3D,E, respectively. |
T125 |
561-662 |
Sentence |
denotes |
The free N protein formed a tetramer through CTD with the NTD freely hanging in solution (Figure 3D). |
T126 |
663-863 |
Sentence |
denotes |
The conformation of the solution was consistent with structures previously reported for other CoV N proteins.33 The N-P3 complex formed a compact hexadecamer with a sunburst configuration (Figure 3E). |
T127 |
864-962 |
Sentence |
denotes |
The CTDs formed a central ring and non-native NTD dimers formed “spikes” protruding from the ring. |
T128 |
963-1135 |
Sentence |
denotes |
Consistent with ligand-induced aggregation, we observed a “blue shift” in the fluorescence spectrum of the full-length MERS-CoV N protein in the presence of P3 (Figure 3F). |
T129 |
1136-1338 |
Sentence |
denotes |
The addition of P3 also delayed N protein thermal denaturation and changed the shape of the denaturation curve, further suggesting that large protein aggregates formed in the presence of P3 (Figure 3G). |
T130 |
1339-1414 |
Sentence |
denotes |
The structure explains how N-NTD dimerization decreased MERS-CoV viability. |
T131 |
1415-1475 |
Sentence |
denotes |
The N protein packages the viral genome into an RNP complex. |
T132 |
1476-1735 |
Sentence |
denotes |
Several models for N-CTD dimer assembly have been proposed for the formation of filamentous RNPs.33 All of the proposed interfaces between N-CTD dimers occurred on the side-faces of the CTD cuboid perpendicular to the proposed RNA-binding surface (Figure 3H). |
T133 |
1736-2027 |
Sentence |
denotes |
Combinatorial use of any region on the side-faces of the CTD dimer cuboid may facilitate manipulation of the RNP length and curvature without obstructing the RNA-binding surface.28,34 However, the SAXS results indicated that N-CTD aggregation occurred on the β-sheet floor of the CTD cuboid. |
T134 |
2028-2214 |
Sentence |
denotes |
For this reason, the RNA-binding surface of the CTD is occluded by the neighboring CTD on the ring and by the non-native NTD dimer making direct contact with the CTD (Figures 3H and S3). |
T135 |
2215-2398 |
Sentence |
denotes |
In addition, the CTD cuboids in the aggregation naturally form a topologically closed octamer, leaving no open ends for further addition of CTD cuboids to form a long filamentous RNP. |
T136 |
2399-2556 |
Sentence |
denotes |
Both the loss of the RNA-binding surface and the inability to incorporate further N protein molecules beyond an octamer may inhibit the formation of the RNP. |
T137 |
2557-2640 |
Sentence |
denotes |
Therefore, P3 may inhibit MERS-CoV RNP formation by inducing N protein aggregation. |