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小鼠多瘤病毒衣壳蛋白与导入蛋白的相互作用是病毒 DNA 高效进入细胞核所必需的。

Interaction of the Mouse Polyomavirus Capsid Proteins with Importins Is Required for Efficient Import of Viral DNA into the Cell Nucleus.

机构信息

Department of Genetics and Microbiology, Faculty of Science, Charles University, Vinicna 5, 12844 Prague 2, Czech Republic.

出版信息

Viruses. 2018 Mar 31;10(4):165. doi: 10.3390/v10040165.

DOI:10.3390/v10040165
PMID:29614718
原文链接:https://pmc.ncbi.nlm.nih.gov/articles/PMC5923459/
Abstract

The mechanism used by mouse polyomavirus (MPyV) overcomes the crowded cytosol to reach the nucleus has not been fully elucidated. Here, we investigated the involvement of importin α/β1 mediated transport in the delivery of MPyV genomes into the nucleus. Interactions of the virus with importin β1 were studied by co-immunoprecipitation and proximity ligation assay. For infectivity and nucleus delivery assays, the virus and its capsid proteins mutated in the nuclear localization signals (NLSs) were prepared and produced. We found that at early times post infection, virions bound importin β1 in a time dependent manner with a peak of interactions at 6 h post infection. Mutation analysis revealed that only when the NLSs of both VP1 and VP2/3 were disrupted, virus did not bind efficiently to importin β1 and its infectivity remarkably decreased (by 80%). Nuclear targeting of capsid proteins was improved when VP1 and VP2 were co-expressed. VP1 and VP2 were effectively delivered into the nucleus, even when one of the NLS, either VP1 or VP2, was disrupted. Altogether, our results showed that MPyV virions can use VP1 and/or VP2/VP3 NLSs in concert or individually to bind importins to deliver their genomes into the cell nucleus.

摘要

小鼠多瘤病毒 (MPyV) 用于克服拥挤的细胞质到达细胞核的机制尚未完全阐明。在这里，我们研究了导入蛋白 α/β1 介导的运输在将 MPyV 基因组递送到细胞核中的作用。通过共免疫沉淀和邻近连接测定研究了病毒与导入蛋白 β1 的相互作用。为了进行感染性和核递呈测定，制备并产生了突变了核定位信号（NLS）的病毒及其衣壳蛋白。我们发现，在感染后早期，病毒粒子以时间依赖性方式与导入蛋白 β1 结合，在感染后 6 小时达到相互作用的峰值。突变分析表明，只有当 VP1 和 VP2/3 的 NLS 都被破坏时，病毒才不能有效地与导入蛋白 β1 结合，其感染性显著降低（80%）。当 VP1 和 VP2 共同表达时，衣壳蛋白的核靶向性得到改善。即使破坏了 NLS 中的一个（VP1 或 VP2），VP1 和 VP2 也能有效地递送到细胞核中。总之，我们的结果表明，MPyV 病毒粒子可以协同或单独使用 VP1 和/或 VP2/VP3 NLS 来结合导入蛋白，将其基因组递送到细胞核中。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/03a1/5923459/811f6637c921/viruses-10-00165-g009.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/03a1/5923459/0dd912feb286/viruses-10-00165-g001.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/03a1/5923459/d99fb83242a0/viruses-10-00165-g002.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/03a1/5923459/421af1fd9ef1/viruses-10-00165-g003.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/03a1/5923459/3141f7623bdf/viruses-10-00165-g004.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/03a1/5923459/ec12e0df085c/viruses-10-00165-g005.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/03a1/5923459/89029ac3eece/viruses-10-00165-g006.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/03a1/5923459/a07583d239d9/viruses-10-00165-g007.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/03a1/5923459/0a80733ed41c/viruses-10-00165-g008.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/03a1/5923459/811f6637c921/viruses-10-00165-g009.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/03a1/5923459/0dd912feb286/viruses-10-00165-g001.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/03a1/5923459/d99fb83242a0/viruses-10-00165-g002.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/03a1/5923459/421af1fd9ef1/viruses-10-00165-g003.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/03a1/5923459/3141f7623bdf/viruses-10-00165-g004.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/03a1/5923459/ec12e0df085c/viruses-10-00165-g005.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/03a1/5923459/89029ac3eece/viruses-10-00165-g006.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/03a1/5923459/a07583d239d9/viruses-10-00165-g007.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/03a1/5923459/0a80733ed41c/viruses-10-00165-g008.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/03a1/5923459/811f6637c921/viruses-10-00165-g009.jpg

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