Assembly of herpes simplex virus capsids using the human cytomegalovirus scaffold protein: critical role of the C terminus
Assembly of herpes simplex virus capsids using the human cytomegalovirus scaffold protein: critical role of the C terminus
复制标题
使用人巨细胞病毒支架蛋白组装单纯疱疹病毒衣壳：C 末端的关键作用
作者：
N. Oien;D. Thomsen;M. Wathen;W. Newcomb;J. Brown;F. Homa
单纯疱疹病毒 (HSV) 1 型衣壳组装的一个重要步骤涉及主要衣壳蛋白 (VP5) 与支架蛋白 C 末端（由 UL26.5 基因编码）的相互作用。 HSV 支架蛋白的最后 12 个残基包含 A-X-X-F-V/A-X-Q-M-M-X-X-R 基序，该基序在其他 α 疱疹病毒中发现的支架蛋白之间是保守的，但在 β 或 γ 疱疹病毒成员中不存在。先前的研究表明，牛疱疹病毒1（α疱疹病毒）UL26.5同源物将在功能上替代HSV UL26.5基因（E.J.Haanes等人，J.Virol.69：7375-7379，1995）。人类巨细胞病毒 (HCMV) 基因组中 UL26.5 基因的同源物是 UL80.5 基因。在这些研究中，我们测试了HCMV UL80.5基因是否会替代我们先前描述的杆状病毒衣壳组装系统中的HSV UL26.5基因(D.R.Thomsen等人，J.Virol.68：2442-2457，1994)。结果表明，(i) 当测试全长或截短的（缺少 C 端 65 个氨基酸）UL80.5 蛋白时，没有组装完整的衣壳； (ii)当UL80.5蛋白的C端65个氨基酸被UL26.5蛋白的C端25个氨基酸取代时，制备了完整的衣壳并检测到UL80.5蛋白与VP5的直接相互作用； (iii)当UL80.5蛋白的最后12个氨基酸的序列从RRIFVA ALNKLE变为RRIFVAAMMKLE时，证明了完整衣壳的组装； (iv)支架蛋白的自相互作用是由成熟切割位点N端的序列介导的； (v) UL26.5和UL80.5蛋白不会共组装成支架结构。结果表明，UL26.5和UL80.5蛋白通过蛋白质N末端序列的自相互作用形成支架，并强调C末端对于支架与形成衣壳壳的蛋白质相互作用的重要性。
An essential step in assembly of herpes simplex virus (HSV) type 1 capsids involves interaction of the major capsid protein (VP5) with the C terminus of the scaffolding protein (encoded by the UL26.5 gene). The final 12 residues of the HSV scaffolding protein contains an A-X-X-F-V/A-X-Q-M-M-X-X-R motif which is conserved between scaffolding proteins found in other alphaherpesviruses but not in members of the beta- or gamma-herpesviruses. Previous studies have shown that the bovine herpesvirus 1 (alphaherpesvirus) UL26.5 homolog will functionally substitute for the HSV UL26.5 gene (E. J. Haanes et al., J. Virol. 69:7375-7379, 1995). The homolog of the UL26.5 gene in the human cytomegalovirus (HCMV) genome is the UL80.5 gene. In these studies, we tested whether the HCMV UL80.5 gene would substitute for the HSV UL26.5 gene in a baculovirus capsid assembly system that we have previously described (D. R. Thomsen et al., J. Virol. 68:2442-2457, 1994). The results demonstrate that (i) no intact capsids were assembled when the full-length or a truncated (missing the C-terminal 65 amino acids) UL80.5 protein was tested; (ii) when the C-terminal 65 amino acids of the UL80.5 protein were replaced with the C-terminal 25 amino acids of the UL26.5 protein, intact capsids were made and direct interaction of the UL80.5 protein with VP5 was detected; (iii) assembly of intact capsids was demonstrated when the sequence of the last 12 amino acids of the UL80.5 protein was changed from RRIFVA ALNKLE to RRIFVAAMMKLE; (iv) self-interaction of the scaffold proteins is mediated by sequences N terminal to the maturation cleavage site; and (v) the UL26.5 and UL80.5 proteins will not coassemble into scaffold structures. The results suggest that the UL26.5 and UL80.5 proteins form a scaffold by self-interaction via sequences in the N termini of the proteins and emphasize the importance of the C terminus for interaction of scaffold with the proteins that form the capsid shell.
DOI：
10.1099/0022-1317-64-2-373
发表时间：
1983-02
期刊：
The Journal of general virology
影响因子：
--
作者：
R. Lafemina;G. Hayward
通讯作者：
R. Lafemina;G. Hayward
DOI：
10.1073/pnas.88.23.10792
发表时间：
1991-12-01
影响因子：
11.1
作者：
WELCH, AR;WOODS, AS;GIBSON, W
通讯作者：
GIBSON, W