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

[1]	OUYANG T, ZHANG X W, LIU X H, et al. Co-Infection of swine with porcine circovirus type 2 and other swine viruses[J]. Viruses, 2019, 11(2):185.
[2]	KARUPPANNAN A K, OPRIESSNIG T. Porcine circovirus type 2 (PCV2) vaccines in the context of current molecular epidemiology[J]. Viruses, 2017, 9(5):99.
[3]	TURLEWICZ-PODBIELSKA H, AUGUSTYNIAK A, POMORSKA-MÓL M. Novel porcine circoviruses in view of lessons learned from porcine circovirus type 2-epidemiology and threat to pigs and other species[J]. Viruses, 2022, 14(2):261.
[4]	LV Q Z, GUO K K, ZHANG Y M. Current understanding of genomic DNA of porcine circovirus type 2[J]. Virus Genes, 2014, 49(1):1-10.
[5]	TIMMUSK S, FOSSUM C, BERG M. Porcine circovirus type 2 replicase binds the capsid protein and an intermediate filament-like protein[J]. J Gen Virol, 2006, 87(Pt 11):3215-3223.
[6]	MEEHAN B M, CREELAN J L, MCNULTY M S, et al. Sequence of porcine circovirus DNA:affinities with plant circoviruses[J]. J Gen Virol, 1997, 78(Pt 1):221-227.
[7]	MANKERTZ A, PERSSON F, MANKERTZ J, et al. Mapping and characterization of the origin of DNA replication of porcine circovirus[J]. J Virol, 1997, 71(3):2562-2566.
[8]	STEINFELDT T, FINSTERBUSCH T, MANKERTZ A. Demonstration of nicking/joining activity at the origin of DNA replication associated with the rep and rep' proteins of porcine circovirus type 1[J]. J Virol, 2006, 80(13):6225-6234.
[9]	KIM C, WANG X D, YU Y H. PARP1 inhibitors trigger innate immunity via PARP1 trapping-induced DNA damage response[J]. Elife, 2020, 9:e60637.
[10]	KO H L, REN E C. Novel poly (ADP-ribose) polymerase 1 binding motif in hepatitis B virus core promoter impairs DNA damage repair[J]. Hepatology, 2011, 54(4):1190-1198.
[11]	NA T Y, KA N L, RHEE H, et al. Interaction of hepatitis B virus X protein with PARP1 results in inhibition of DNA repair in hepatocellular carcinoma[J]. Oncogene, 2016, 35(41):5435-5445
[12]	SUN R J, SUN S Q, ZHANG Y K, et al. PCV2 induces reactive oxygen species to promote nucleocytoplasmic translocation of the viral DNA binding protein HMGB1 to enhance its replication[J]. J Virol, 2020, 94(13):e00238-20.
[13]	CHEUNG A K. Transcriptional analysis of porcine circovirus type 2[J]. Virology, 2003, 305(1):168-180.
[14]	CHEUNG A K. Porcine circovirus:transcription and DNA replication[J]. Virus Res, 2012, 164(1-2):46-53.
[15]	FINSTERBUSCH T, STEINFELDT T, DOBERSTEIN K, et al. Interaction of the replication proteins and the capsid protein of porcine circovirus type 1 and 2 with host proteins[J]. Virology, 2009, 386(1):122-131.
[16]	MANKERTZ A. Molecular interactions of porcine circoviruses type 1 and type 2 with its host[J]. Virus Res, 2012, 164(1-2):54-60.
[17]	ZHANG X, ZHOU J Y, WU Y P, et al. Differential proteome analysis of host cells infected with porcine circovirus type 2[J]. J Proteome Res, 2009, 8(11):5111-5119.
[18]	HE J, LENG C L, PAN J Z, et al. Identification of lncRNAs involved in PCV2 infection of PK-15 cells[J]. Pathogens, 2020, 9(6):479.
[19]	CHEN X X, REN F, HESKETH J, et al. Interaction of porcine circovirus type 2 replication with intracellular redox status in vitro[J]. Redox Rep, 2013, 18(5):186-192.
[20]	ZHU B L, ZHOU Y S, XU F, et al. Porcine circovirus type 2 induces autophagy via the AMPK/ERK/TSC2/mTOR signaling pathway in PK-15 cells[J]. J Virol, 2012, 86(22):12003-12012.
[21]	ZHOU Y S, QI B Z, GU Y X, et al. Porcine circovirus 2 deploys PERK pathway and GRP78 for its enhanced replication in PK-15 cells[J]. Viruses, 2016, 8(2):56.
[22]	张辉, 唐青海, 危艳武, 等. 与猪圆环病毒2型基因组茎环结构相互作用的宿主细胞蛋白的筛选[J]. 中国兽医科学, 2013, 43(12):1211-1216.ZHANG H, TANG Q H, WEI Y W, et al. Screening of host proteins interacting with porcine circovirus type 2 genome stem-loop structure[J]. Chinese Veterinary Science, 2013, 43(12):1211-1216. (in Chinese)
[23]	ZHU H F, ZHENG C F. When PARPs meet antiviral innate immunity[J]. Trends Microbiol, 2021, 29(9):776-778.
[24]	CHAMBERS B S, HEATON B E, RAUSCH K, et al. DNA mismatch repair is required for the host innate response and controls cellular fate after influenza virus infection[J]. Nat Microbiol, 2019, 4(11):1964-1977.
[25]	OHASHI E, TSURIMOTO T. Functions of multiple clamp and clamp-loader complexes in eukaryotic DNA replication[M]//MASAI H, FOIANI M. DNA Replication. Singapore:Springer, 2017:135-162.
[26]	MAYER D, MOLAWI K, MARTÍNEZ-SOBRIDO L, et al. Identification of cellular interaction partners of the influenza virus ribonucleoprotein complex and polymerase complex using proteomic-based approaches[J]. J Proteome Res, 2007, 6(2):672-682.
[27]	BORTZ E, WESTERA L, MAAMARY J, et al. Host- and strain-specific regulation of influenza virus polymerase activity by interacting cellular proteins[J]. Mbio, 2011, 2(4):e00151-11.
[28]	HAGEN T M, HUANG S, CURNUTTE J, et al. Extensive oxidative DNA damage in hepatocytes of transgenic mice with chronic active hepatitis destined to develop hepatocellular carcinoma[J]. Proc Natl Acad Sci U S A, 1994, 91(26):12808-12812.
[29]	ZHANG Z, HILDEBRANDT E F, SIMBULAN-ROSENTHAL C M, et al. Sequence-specific binding of poly (ADP-ribose) polymerase-1 to the human T cell leukemia virus type-I tax responsive element[J]. Virology, 2002, 296(1):107-116.
[30]	OHSAKI E, UEDA K, SAKAKIBARA S, et al. Poly (ADP-ribose) polymerase 1 binds to kaposi's sarcoma-associated herpesvirus (KSHV) terminal repeat sequence and modulates KSHV replication in latency[J]. J Virol, 2004, 78(18):9936-9946.
[31]	WANG Y, LI H, TANG Q Y, et al. Kaposi's sarcoma-associated herpesvirus ori-Lyt-dependent DNA replication:involvement of host cellular factors[J]. J Virol, 2008, 82(6):2867-2882.
[32]	KIM M Y, ZHANG T, KRAUS W L. Poly (ADP-ribosyl) ation by PARP-1:‘PAR-laying’ NAD+ into a nuclear signal[J]. Genes Dev, 2005, 19(17):1951-1967.
[33]	KRIETSCH J, ROULEAU M, PIC É, et al. Reprogramming cellular events by poly (ADP-ribose)-binding proteins[J]. Mol Aspects Med, 2013, 34(6):1066-1087.

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

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