The SV40 Capsid Is Stabilized by a Conserved Pentapeptide Hinge of the Major Capsid Protein VP1

被引:14
作者
Ben-nun-Shaul, Orly [2 ]
Bronfeld, Hagit [2 ]
Reshef, Dan [1 ]
Schueler-Furman, Ora [1 ]
Oppenheim, Ariella [2 ]
机构
[1] Hebrew Univ Jerusalem, Hadassah Med Sch, Dept Mol Genet & Biotechnol, IL-91120 Jerusalem, Israel
[2] Hebrew Univ Jerusalem, Hadassah Med Sch, Dept Hematol, IL-91120 Jerusalem, Israel
基金
美国国家卫生研究院;
关键词
SV40; major capsid protein VP1; site-directed mutagenesis; computational binding prediction; protein structure-function; SIMIAN-VIRUS-40; VP1; PACKAGING SIGNAL; SIMIAN VIRUS-40; VISUALIZATION; RESOLUTION; COMPLEXES; SES;
D O I
10.1016/j.jmb.2008.11.034
中图分类号
Q5 [生物化学]; Q7 [分子生物学];
学科分类号
071010 ; 081704 ;
摘要
The simian virus 40 (SV40) outer shell is composed of 72 pentamers of VP1. The core of the VP1 monomer is a beta-barrel with jelly-roll topology and extending N- and C-terminal arms. A pentapeptide hinge, KNPYP, tethers the C-arm to the VP1 beta-barrel core. The five C-arms that extend from each pentamer insert into the neighbouring pentamers, tying them together through different types of interactions. In the mat-Lire virion, this element adopts either of six conformations according to their location in the capsid. We found that the hinge is conserved among 16 members of the Polyomaviridae, attesting to its importance in capsid assembly and/or structure. We have used site-directed mutagenesis to gain an understanding into the structural requirements of this element: Y299 was changed to A, F, and T, and P300 to A and G. The mutants showed reduction in viability to varying degrees. Unexpectedly, assembly was reduced only to a small extent. However, the data showed that the mutants were highly unstable. The largest effect was observed for mutations of P300, indicating a role of the proline in the virion structure. P300G was more unstable than P300A, indicating a requirement for rigidity of the pentapeptide hinge. Y299T and Y299A were more defective in viability than Y299F, highlighting the importance of an aromatic ring at this position. Structural inspection showed that this aromatic ring contacts C-arms of neighbouring pentamers. Computational modelling predicted loss of stability of the Y mutants in concordance with the experimental results. This study provides insights into the structural. details of the pentapeptide hinge that are responsible for capsid stability. (c) 2008 Elsevier Ltd. All rights reserved.
引用
收藏
页码:1382 / 1391
页数:10
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