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N6-甲基腺苷修饰细小病毒编码的小非编码 RNA 通过募集 Y 家族 DNA 聚合酶促进病毒 DNA 复制。

N6-methyladenosine modification of a parvovirus-encoded small noncoding RNA facilitates viral DNA replication through recruiting Y-family DNA polymerases.

机构信息

Department of Microbiology, Molecular Genetics and Immunology, University of Kansas Medical Center, Kansas City, KS 66160.

Department of Medicinal Chemistry, University of Kansas, Lawrence, KS 66045.

出版信息

Proc Natl Acad Sci U S A. 2024 Jun 18;121(25):e2320782121. doi: 10.1073/pnas.2320782121. Epub 2024 Jun 14.

DOI:10.1073/pnas.2320782121
PMID:38875150
原文链接:https://pmc.ncbi.nlm.nih.gov/articles/PMC11194592/
Abstract

Human bocavirus 1 (HBoV1) is a human parvovirus that causes lower respiratory tract infections in young children. It contains a single-stranded (ss) DNA genome of ~5.5 kb that encodes a small noncoding RNA of 140 nucleotides known as bocavirus-encoded small RNA (BocaSR), in addition to viral proteins. Here, we determined the secondary structure of BocaSR in vivo by using DMS-MaPseq. Our findings reveal that BocaSR undergoes N6-methyladenosine (m6A) modification at multiple sites, which is critical for viral DNA replication in both dividing HEK293 cells and nondividing cells of the human airway epithelium. Mechanistically, we found that m6A-modified BocaSR serves as a mediator for recruiting Y-family DNA repair DNA polymerase (Pol) η and Pol κ likely through a direct interaction between BocaSR and the viral DNA replication origin at the right terminus of the viral genome. Thus, this report represents direct involvement of a viral small noncoding RNA in viral DNA replication through m6A modification.

摘要

人博卡病毒 1（HBoV1）是一种人类细小病毒，可导致幼儿下呼吸道感染。它包含一个约 5.5 kb 的单链（ss）DNA 基因组，编码一个 140 个核苷酸的小非编码 RNA，称为博卡病毒编码的小 RNA（BocaSR），此外还有病毒蛋白。在这里，我们通过 DMS-MaPseq 确定了 BocaSR 的体内二级结构。我们的研究结果表明，BocaSR 在多个位点发生 N6-甲基腺苷（m6A）修饰，这对于病毒 DNA 在有丝分裂的 HEK293 细胞和非分裂的人呼吸道上皮细胞中的复制至关重要。从机制上讲，我们发现 m6A 修饰的 BocaSR 通过 BocaSR 与病毒基因组右端病毒 DNA 复制起点之间的直接相互作用，作为招募 Y 家族 DNA 修复 DNA 聚合酶（Pol）η和 Pol κ 的介质。因此，本报告代表了病毒小非编码 RNA 通过 m6A 修饰直接参与病毒 DNA 复制。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/a2df/11194592/22c3c5741fee/pnas.2320782121fig08.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/a2df/11194592/78ba029c50b1/pnas.2320782121fig01.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/a2df/11194592/4115b1a22c6e/pnas.2320782121fig02.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/a2df/11194592/9a7c9a6c7db8/pnas.2320782121fig03.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/a2df/11194592/105a0ecb4c28/pnas.2320782121fig04.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/a2df/11194592/9b6dd50f1d97/pnas.2320782121fig05.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/a2df/11194592/9bbda1819f55/pnas.2320782121fig06.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/a2df/11194592/eb7403629a4a/pnas.2320782121fig07.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/a2df/11194592/22c3c5741fee/pnas.2320782121fig08.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/a2df/11194592/78ba029c50b1/pnas.2320782121fig01.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/a2df/11194592/4115b1a22c6e/pnas.2320782121fig02.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/a2df/11194592/9a7c9a6c7db8/pnas.2320782121fig03.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/a2df/11194592/105a0ecb4c28/pnas.2320782121fig04.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/a2df/11194592/9b6dd50f1d97/pnas.2320782121fig05.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/a2df/11194592/9bbda1819f55/pnas.2320782121fig06.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/a2df/11194592/eb7403629a4a/pnas.2320782121fig07.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/a2df/11194592/22c3c5741fee/pnas.2320782121fig08.jpg

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