A single amino acid substitution in the cyclin D binding domain of the infected cell protein no. 0 abrogates the neuroinvasiveness of herpes simplex virus without affecting its ability to replicate

作     者：Van Sant, C Kawaguchi, Y Roizman, B 

作者机构：Univ Chicago Marjorie B Kovler Viral Oncol Labs Chicago IL 60637 USA 

出 版 物：《PROCEEDINGS OF THE NATIONAL ACADEMY OF SCIENCES OF THE UNITED STATES OF AMERICA》 (美国国家科学院汇刊)

年 卷 期：1999年第96卷第14期

页      面：8184-8189页

核心收录：

学科分类：07[理学] 08[工学] 

基　　金：NCI NIH HHS [R01 CA078766, R37 CA078766, CA78766, CA71933, CA47451, P01 CA071933] Funding Source: Medline NIGMS NIH HHS [T32 GM007183] Funding Source: Medline 

主　　题：氨基酸取代 天冬氨酸 结合部位 细胞 培养的 中枢神经系统/病毒学 黑长尾猴 密码子 接触抑制 细胞周期蛋白D3 细胞周期蛋白类/代谢 基因组 病毒 HeLa细胞 疱疹病毒1型 人/遗传学 疱疹病毒1型 人/致病力 疱疹病毒1型 人/生理学 即早蛋白质类/化学 即早蛋白质类/代谢 诱变 定点 磷酰化 重组蛋白质类/化学 重组蛋白质类/代谢 重组 遗传 视网膜母细胞瘤蛋白质/代谢 皮肤/病毒学 肿瘤细胞 培养的 泛素蛋白连接酶类 Vero细胞 毒力 病毒复制 动物 人类 小鼠 兔 

摘      要：The infected cell protein no, 0 (ICP0) of herpes simplex virus 1 is a promiscuous transactivator shown to enhance the expression of genes introduced into cells by infection or transfection. The protein interacts with several viral and cellular proteins. Earlier studies have shown that ICP0 binds and stabilizes cyclin D3 but does interfere with the phosphorylation of retinoblastoma protein, its major function. Cyclin D3 plays a key role in the transition from GI to S phase. To define the role of cyclin D3 in productive infection, the ICP0 binding site for cyclin D3 was mapped and mutagenized by substitution of aspartic acid codon 199 with the alanine codon. We report that the substitution precluded the interaction of this protein with cyclin D3 in the yeast two-hybrid system and the stabilization of cyclin D3 in infected cells, A recombinant virus carrying this mutation could not be differentiated from wild-type parent with respect to replication in dividing cells but yielded 10-fold less progeny from infected resting cells and serum-deprived or contact-inhibited human fibroblasts, In mice, the mutant was only slightly less pathogenic than the wild-type parent by intracerebral route but was significantly less neuroinvasive after peripheral inoculation. Replacement of the mutated amino acid with aspartic acid restored wild-type phenotype. Stabilization of cyclin D3 therefore is linked to higher:virus yields in nondividing cells and potentially higher virulence in experimental and natural hosts, One function of ICP0 is to scavenge the cell for proteins that could bolster viral replication.