Analysis of mouse polyomavirus mutants with lesions in the minor capsid proteins.

Analysis of mouse polyomavirus mutants with lesions in the minor capsid proteins.
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分析具有次要衣壳蛋白损伤的小鼠多瘤病毒突变体。

DOI:
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发表时间:
2002
影响因子:
3.8
通讯作者:
J. Forstová
J. Forstová
中科院分区:
医学3区
文献类型:
--
作者:
Petra Mannová;D. Liebl;N. Krauzewicz;Anna Fejtová;J. Štokrová;Z. Palková;B. Griffin;J. Forstová

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多瘤病毒突变体E、Q和H分别用谷氨酸、谷氨酰胺或组氨酸取代N-末端甘氨酸残基,获得表达非肉豆蔻化VP2的突变体。还制备了VP2或VP3翻译起始密码子突变的病毒。所有突变的基因组,当被转移到小鼠宿主细胞中时,都会产生病毒颗粒。VP2和VP3病毒的感染性,以表达病毒抗原的细胞数量衡量,显著降低,表明感染早期存在缺陷。相反,VP2上缺少多年生花柱部分并没有实质上影响病毒感染的早期步骤。在一个生命周期中,野生型(Wt)病毒和E或Q MYR病毒在表达早期或晚期病毒抗原的细胞数量上没有差异。此外,没有检测到病毒DNA复制的延迟。然而,当细胞在培养中停留更长时间时,使用突变基因组而不是wt基因组时,以典型病毒爆发衡量的感染细胞数量要低得多。原位细胞分离研究显示,病毒颗粒与宿主细胞结构的相互作用存在差异。突变体的侵染性不仅受VP2上肉豆蔻基缺失的影响,而且在更大程度上还受到N末端氨基酸组成的改变的影响。
Polyomavirus mutants E, Q and H, expressing non-myristylated VP2, were generated by replacing the N-terminal glycine residue with glutamic acid, glutamine or histidine, respectively. Viruses mutated in either VP2 or VP3 translation initiation codons were also prepared. All mutated genomes, when transfected into murine host cells, gave rise to viral particles. Infectivity of VP2- and VP3- viruses, as measured by the number of cells expressing viral antigens, was dramatically diminished, indicative of defects in the early stages of infection. In contrast, the absence of a myristyl moiety on VP2 did not substantially affect the early steps of virus infection. No differences in numbers of cells expressing early or late viral antigens were observed between wild-type (wt) and E or Q myr- viruses during the course of a life cycle. Furthermore, no delay in virus DNA replication was detected. However, when cells were left for longer in culture, the number of infected cells, measured by typical virus bursts, was much lower when mutant rather than wt genomes were used. In situ, cell fractionation studies revealed differences in the interaction of viral particles with host cell structures. The infectivity of mutants was affected not only by loss of the myristyl group on VP2, but also, and to a greater extent, by alterations of the N-terminal amino acid composition.
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