E3泛素连接酶RNF185对A亚群禽白血病病毒体外复制的影响

doi:10.16431/j.cnki.1671-7236.2022.11.027

Abstract

Abstract: 【Objective】 Ring finger protein 185 (RNF185) is a member of Ring family E3 ubiquitin ligases.This study was aimed to explore the effect of chicken-derived RNF185 on the replication of Avian leukemia virus subgroup A (ALV-A) in vitro, and study the effect of chicken-derived RNF185 on the pathogenicity and mediated tumorigenic mechanism of ALV-A.【Method】 Referring to the sequence of RNF185 gene in chicken (accession No.:NP_001007841) published in GenBank, RNF185 overexpression primers and RNF185 single-stranded shRNA were designed, respectively.The RNF185 gene and double-stranded shRNA were synthesized by PCR and cloned into eukaryotic expression vectors psi-flag and interference vector pLKO.1-GFP, respectively.The constructed recombinant plasmids were verified by PCR, double restriction digestion and sequencing, and the CCK-8 kit was used to detect the effect of the recombinant plasmid on the viability of DF-1 cells.10⁴ tissue culture infective dose 50% (TCID₅₀) ALV-A (rHB2015012) virus solution was inoculated into DF-1 cells after RNF185 overexpression and interference, respectively, the cells were harvested at 3, 4 and 5 d after inoculation, and the viral load of ALV-A was detected by Western blotting.【Result】 The RNF185 gene was successfully amplified by PCR and the single-stranded shRNA was annealed to form double-stranded shRNA.The recombinant plasmids psi-flag-RNF185, pLKO.1-RNF185-1 and pLKO.1-RNF185-2 were successfully constructed.Western blotting results showed that the constructed overexpression and interference plasmid could stably express and interfere with RNF185 in DF-1 cells.The CCK-8 results indicated that overexpression and interference of RNF185 did not affect the activity of DF-1 cells.Overexpression of RNF185 could promote the replication of ALV-A in DF-1 cells, and interfere with the expression of RNF185 could inhibit the replication of ALV-A in DF-1 cells.【Conclusion】 This study constructed an overexpression plasmid and an interference plasmid that could highly express and interfere with RNF185 in DF-1 cells.RNF185 had no significant effect on the proliferation of DF-1 cells, confirming that RNF185 could promote the replication of ALV-A (rHB2015012) in vitro.

Key words: ring finger protein 185 (RNF185); Avian leukemia virus subgroup A (ALV-A); virus replication

CLC Number:

S852.65⁺7

LIU Jing, CHEN Xueyang, ZENG Yang, YANG Yuying. Effect of E3 Ubiquitin Ligase RNF185 on Replication of Avian Leukemia Virus Subgroup A in vitro[J]. China Animal Husbandry and Veterinary Medicine, 2022, 49(11): 4364-4371.

References

[1] QU Y, LIU L, NIU Y, et al.Viral proliferation and expression of tumor-related gene in different chicken embryo fibroblasts infected with different tumorigenic phenotypes of Avian leukosis virus subgroup J[J].Poultry Science, 2016, 95(10):2383-2390.
[2] LIANG X Y, GU Y F, CHEN X Y, et al.Identification and characterization of a novel natural recombinant Avian leucosis virus from Chinese indigenous chicken flock[J].Virus Genes, 2019, 55(5):726-733.
[3] PANG Y, ZHOU D F, XUE J W, et al.Interplay between CTHRC1 and the SU protein of Avian leukosis virus subgroup J (ALV-J) facilitates viral replication[J].Virus Research, 2019, 264:32-39.
[4] 王鑫, 赵鹏, 崔治中.我国地方品种鸡分离到的一个禽白血病病毒新亚群的鉴定[J].病毒学报, 2012, 28(6):609-614. WANG X, ZHAO P, CUI Z Z.Identification of a new subgroup of Avian leukosis virus isolated from Chinese indigenous chicken breeds[J].Chinese Journal of Virology, 2012, 28(6):609-614.(in Chinese)
[5] 崔治中, 郭惠君, 孙淑红.鸡白血病的流行现状和防制对策[J].中国兽药杂志, 2009, 43(10):37-41. CUI Z Z, GUO H J, SUN S H.Prevalence situation and prevention and control of avian leukosis[J].Chinese Journal of Veterinary Drug, 2009, 43(10):37-41.(in Chinese)
[6] PAYNE L N, NAIR V.The long view:40 years of avian leukosis research[J].Avian Pathology, 2012, 41(1):11-19.
[7] 崔治中.种鸡场禽白血病防控和净化技术方案[J].中国家禽, 2015, 37(23):1-7. CUI Z Z.Technical scheme on prevention and control of avian leucosis in breeding chicken farm[J].China Poultry, 2015, 37(23):1-7.(in Chinese)
[8] POPOVIC D, VUCIC D, DIKIC I.Ubiquitination in disease pathogenesis and treatment[J].Nature Medicine, 2014, 20(11):1242-1253.
[9] TOMA-FUKAI S, SHIMIZU T.Structural diversity of ubiquitin E3 ligase[J].Molecules (Basel, Switzerland), 2021, 26(21):6682.
[10] SHAID S, BRANDTS C H, SERVE H, et al.Ubiquitination and selective autophagy cell death and differentiation[J].Cell Death and Differentiation, 2013, 20(1):21-30.
[11] ADAM H, RICARDO R.To TRIM or not to TRIM:The balance of host-virus interactions mediated by the ubiquitin system[J].The Journal of General Virology, 2019, 100(12):1641-1662.
[12] QIU D P, WANG Q, WANG Z D, et al.RNF185 modulates JWA ubiquitination and promotes gastric cancer metastasis[J].Biochimica et Biophysica Acta.Molecular Basis of Disease, 2018, 1864(5 Pt A):1552-1561.
[13] KIM S Y, HERBST A, TWORKOWSKI K A, et al.Skp2 regulates Myc protein stability and activity[J].Molecular Cell, 2003, 11(5):1177-1188.
[14] ZHANG Z S, HU W G, LI B Q, et al.Comparison of viremia, Cloacal virus shedding, antibody responses and pathological lesions in adult chickens, quails, and pigeons infected with ALV-A[J].Scientific Reports, 2019, 9(1):3027.
[15] MAO Y Q, SU Q, LI J P, et al.Avian leukosis virus contamination in live vaccines:A retrospective investigation in China[J].Veterinary Microbiology, 2020, 246(7):108712.
[16] DAI M, FENG M, LIU D, et al.Development and application of SYBR Green Ⅰ Real-time PCR assay for the separate detection of subgroup J Avian leukosis virus and multiplex detection of Avian leukosis virus subgroups A and B[J].Journal of Virology, 2015, 12:52.
[17] BO Z, ZHANG L, LEI C, et al.The ubiquitin ligase RNF5 regulates antiviral responses by mediating degradation of the adaptor protein MITA[J].Immunity, 2009, 30(3):397-407.
[18] KHATEB A, DESHPANDE A, FENG Y, et al.The ubiquitin ligase RNF5 determines acute myeloid leukemia growth and susceptibility to histone deacetylase inhibitors[J].Nature Communications, 2021, 12(1):5397.
[19] GAO Y, XUAN C, JIN M, et al.Ubiquitin ligase RNF5 serves an important role in the development of human glioma[J].Oncology Letters, 2019, 18(5):4659-4666.
[20] FEI T, WANG B, LI N, et al.RNF185, a novel mitochondrial ubiquitin E3 ligase, regulates autophagy through interaction with BNIP1[J].PLoS One, 2011, 6(9):e24367.
[21] WELCKER M, ORIAN A, JIN J, et al.The Fbw7 tumor suppressor regulates glycogen synthase kinase 3 phosphorylation-dependent c-Myc protein degradation[J].Proceedings of the National Academy of Sciences of the United States of America, 2004, 101(24):9085-9090.
[22] ZHANG X, YAN Y, LIN W, et al.Circular RNA Vav3 sponges gga-miR-375 to promote epithelial-mesenchymal transition[J]. RNA Biology, 2019, 16(1):118-132.
[23] REN C, YU M, ZHANG Y, et al.Avian leukosis virus subgroup J promotes cell proliferation and cell cycle progression through miR-221 by targeting CDKN1B[J].Virology, 2018, 519:121-130.
[24] LI Z, LUO Q, XU H, et al.miR-34b-5p suppresses melanoma differentiation-associated gene 5(MDA5) signaling pathway to promote Avian leukosis virus subgroup J (ALV-J)-infected cells proliferaction and ALV-J replication[J].Frontiers in Cellular and Infection Microbiology, 2017, 7:17.
[25] 张晨, 左其生, 邹艺琛, 等.鸡Ash2l基因携带Flag标签的真核融合表达载体构建及表达分析[J].农业生物技术学报, 2021, 29(6):1161-1168. ZHANG C, ZUO Q S, ZOU Y C, et al.Construction and expression analysis of eukaryotic fusion expression vector carrying flag tag of Ash2l gene in chicken (Gallus gallus)[J].Journal of Agricultural Biotechnology, 2021, 29(6):1161-1168.(in Chinese)
[26] 刘容秀, 凌莹, 喜琴琴, 等.ANXA2真核表达载体的构建[J].现代畜牧兽医, 2019, 8:17-21. LIU R X, LING Y, XI Q Q, et al.Optimization of the construction process of eukaryotic expression vector[J].Modern Journal of Animal Husbandry and Veterinary Medicine, 2019, 8:17-21.(in Chinese)

Effect of E3 Ubiquitin Ligase RNF185 on Replication of Avian Leukemia Virus Subgroup A in vitro

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