PCV2复制起始区DNA互作蛋白的筛选及PARP1蛋白对病毒复制的调控

doi:10.11843/j.issn.0366-6964.2022.10.023

Abstract

Abstract: The purpose of this study was to gain a cell protein profile that interact with the porcine circovirus type 2 (PCV2) replication initiation region (Ori) in PK-15 cells and the regulation of PARP1 protein on PCV2 replication was preliminarily investigated. To gain a protein profile that interact with the PCV2 Ori in PK-15 cells, we performed a biotinylated DNA-protein pull down assay in conjunction with LC-MS/MS analysis; GO analysis and KEGG pathway enrichment analysis were performed and the protein interaction network was mapped; subsequently, we constructed eukaryotic plasmids of PARP1 and viral protein Rep, and the interaction between PARP1 protein and Ori region was verified by DNA pull down and IP experiments; Co-IP assay was used to detect the binding of PARP1 protein to Rep proteins; finally, the effect of over-expression or silencing of PARP1 protein on virus replication was detected. A total of 130 host proteins were identified, in which 45 were related to DNA function, mainly involved in host DNA damage repair and replication function; the interaction between PARP1 protein and PCV2 genome Ori or Rep protein was verified; after siRNA silenced the expression of PARP1 protein, the copy number of PCV2 genome was significantly decreased, and PARP1 protein was overexpressed instantly, and the copy number of PCV2 genome was significantly increased. In conclusion, the replication initiation of PCV2 in host cells involves the repair of host DNA damage repair and replication function. PARP1 protein could bind to Ori and Rep proteins of PCV2 and promote the replication of PCV2, which lays a foundation for the study of drug targets of PCV2 and the replication initiation of PCV2.

Key words: porcine circovirus type 2, replication initiation region, host interaction proteins, poly (ADP-ribose) polymerase 1, DNA damage repair

CLC Number:

S852.659.2

JIA Hanxiao, YUAN Honggen, LI Zhen, GUAN Shuaiyin, ZHANG Jinhua, SONG Yunfeng. Screening of Proteins Interacting with PCV2 Replication Initiation Region and Study on the Regulation of Virus Replication by PARP1 Protein[J]. Acta Veterinaria et Zootechnica Sinica, 2022, 53(10): 3511-3521.

References 33

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Screening of Proteins Interacting with PCV2 Replication Initiation Region and Study on the Regulation of Virus Replication by PARP1 Protein

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