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SYNCRIP 通过 NS1 mRNA 的选择性剪接促进 NS2 mRNA 的形成，从而促进猪细小病毒病毒 DNA 的复制。

SYNCRIP facilitates porcine parvovirus viral DNA replication through the alternative splicing of NS1 mRNA to promote NS2 mRNA formation.

机构信息

College of Veterinary Medicine, Northwest A&F University, Yangling, China.

出版信息

Vet Res. 2021 May 25;52(1):73. doi: 10.1186/s13567-021-00938-6.

DOI:10.1186/s13567-021-00938-6
PMID:34034820
原文链接:https://pmc.ncbi.nlm.nih.gov/articles/PMC8152309/
Abstract

Porcine Parvovirus (PPV), a pathogen causing porcine reproductive disorders, encodes two capsid proteins (VP1 and VP2) and three nonstructural proteins (NS1, NS2 and SAT) in infected cells. The PPV NS2 mRNA is from NS1 mRNA after alternative splicing, yet the corresponding mechanism is unclear. In this study, we identified a PPV NS1 mRNA binding protein SYNCRIP, which belongs to the hnRNP family and has been identified to be involved in host pre-mRNA splicing by RNA-pulldown and mass spectrometry approaches. SYNCRIP was found to be significantly up-regulated by PPV infection in vivo and in vitro. We confirmed that it directly interacts with PPV NS1 mRNA and is co-localized at the cytoplasm in PPV-infected cells. Overexpression of SYNCRIP significantly reduced the NS1 mRNA and protein levels, whereas deletion of SYNCRIP significantly reduced NS2 mRNA and protein levels and the ratio of NS2 to NS1, and further impaired replication of the PPV. Furthermore, we found that SYNCRIP was able to bind the 3'-terminal site of NS1 mRNA to promote the cleavage of NS1 mRNA into NS2 mRNA. Taken together, the results presented here demonstrate that SYNCRIP is a critical molecule in the alternative splicing process of PPV mRNA, while revealing a novel function for this protein and providing a potential target of antiviral intervention for the control of porcine parvovirus disease.

摘要

猪细小病毒（PPV）是一种引起猪繁殖障碍的病原体，在感染细胞中编码两种衣壳蛋白（VP1 和 VP2）和三种非结构蛋白（NS1、NS2 和 SAT）。PPV 的 NS2 mRNA 是 NS1 mRNA 通过选择性剪接产生的，但对应的机制尚不清楚。在本研究中，我们鉴定了一个 PPV NS1 mRNA 结合蛋白 SYNCRIP，它属于 hnRNP 家族，已通过 RNA 下拉和质谱方法鉴定为参与宿主前体 mRNA 剪接。研究发现，SYNCRIP 在体内和体外 PPV 感染时显著上调。我们证实它直接与 PPV NS1 mRNA 相互作用，并在 PPV 感染细胞的细胞质中共定位。SYNCRIP 的过表达显著降低了 NS1 mRNA 和蛋白水平，而 SYNCRIP 的缺失则显著降低了 NS2 mRNA 和蛋白水平以及 NS2 与 NS1 的比值，进一步抑制了 PPV 的复制。此外，我们发现 SYNCRIP 能够结合 NS1 mRNA 的 3'-末端位点，促进 NS1 mRNA 切割成 NS2 mRNA。综上所述，本研究结果表明，SYNCRIP 是 PPV mRNA 选择性剪接过程中的关键分子，同时揭示了该蛋白的新功能，并为控制猪细小病毒病提供了抗病毒干预的潜在靶点。

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