Point mutation in calcium-binding domain of mouse polyomavirus VP1 protein does not prevent virus-like particle formation, but changes VP1 interactions with Saccharomyces cerevisiae cell structures
Point mutation in calcium-binding domain of mouse polyomavirus VP1 protein does not prevent virus-like particle formation, but changes VP1 interactions with Saccharomyces cerevisiae cell structures
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DOI：
10.1016/j.femsyr.2004.10.012
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发表时间：
2005-02-01
影响因子：
3.2
通讯作者：
Forstová, J
中科院分区：
文献类型：
--
作者：
Adamec, T;Palková, Z;Forstová, J
在酿酒酵母和杆状病毒表达系统中表达了主要结构蛋白VP1的小鼠多瘤病毒基因VP1(ALA)。令人惊讶的是，VP1(ALA)在酵母和昆虫细胞的细胞核中形成病毒样颗粒(VLP)。酿酒酵母产生的VP1(Ala)-VLP是不稳定的，与野生型VP1(VP1(Wt))-VLP不同，它们在纯化过程和储存过程中会分解。与VP1(Wt)不同，VP1(Ala)不与酵母有丝分裂纺锤体相互作用。然而，野生型和突变型VP1都抑制酵母细胞的生长。这种抑制作用是cAMP依赖的。VP1(丙氨酸)和VP1(Wt)-VLP在昆虫细胞中的产生也显示出它们与细胞蛋白相互作用的不同(S)。因此，VP1钙离子口袋的突变改变了VLP的稳定性和表面构象，而不是改变了VP1的自组装能力。(C)2004年欧洲微生物学会联合会。爱思唯尔出版，版权所有。
The mouse polyomavirus gene for the major structural protein, VP1, with point mutation in the calcium-binding pocket (VP1(Ala)), was expressed in Saccharomyces cerevisiae and in a baculovirus expression system. Surprisingly, VP1(Ala), forms virus-like particles (VLPs) in nuclei of both yeast and insect cells. VP1(Ala)-VLPs produced in S. cerevisiae are unstable and, unlike wild-type VP1 (VP1(wt))-VLPs, they disassemble during the purification procedure and storage. In contrast to VP1(wt), VP1(Ala) does not interact with the yeast mitotic spindle. Nevertheless, both wild-type and mutated VP1 inhibit yeast cell growth. The inhibition is cAMP-dependent. The production of VP1(Ala) and VP1(wt)-VLPs in insect cells also revealed differences in their interactions with cellular protein(s). Thus, the mutation in the VP1 calcium pocket alters the stability and surface conformation of VLPs rather than the ability of VP1 to self-assemble. (C) 2004 Federation of European Microbiological Societies. Published by Elsevier B.V. All rights reserved.