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斜纹夜蛾核多角体病毒假定抗凋亡基因iap4的鉴定与功能分析

Identification and functional analysis of the putative anti-apoptotic gene iap4 of Spodoptera litura nucleopolyhedrovirus.

作者信息

Yu Qian

机构信息

College of Light Industry and Food Science, Zhongkai University of Agiculture and Engineering, Guangzhou, People's Republic of China.

出版信息

Virus Genes. 2010 Oct;41(2):298-304. doi: 10.1007/s11262-010-0513-4. Epub 2010 Jul 13.

Abstract

Baculoviruses possess two types of anti-apoptotic genes, p35 and inhibitor of apoptosis (iap). In this study, we cloned a novel IAP family member, Splt-IAP4, from Spodoptera litura nucleopolyhedrovirus (SpltNPV). ScanProsite analysis of Splt-IAP4 amino acid sequence show that it contain a baculovirus IAP repeat (BIR) motif at the N-terminal portion from 18 to 61 amino acid and a Zinc finger RING motif near its C-terminus. Baculovirus IAPs could be divided into five types from IAP1 to IAP5 according to amino acid sequence homology, so the Splt-IAP4 was grouped into IAP4 because of containing a single BIR motif. Alignment of the six IAP4s in sequenced baculovirus genomes indicated that the Splt-IAP4 gene shared 74% amino acid sequence similarity with Spodoptera littoralis NPV (SpliNPV) IAP4 higher than other baculovirus IAP4 proteins, suggesting evolutionary conservation. In transient expression assay, Splt-IAP4 was not able to suppress apoptosis induced by a mutant AcMNPV deficient in p35 (vAcAnh) infection and rescue the mutant virus replication from apoptosis in Sf-9 cells. Cell viabilities analysis showed that Splt-IAP4 could not inhibit or delay the apoptosis of Sf-9 cells induced by vAcAnh infection. In summary, Splt-IAP4 had no anti-apoptotic function during vAcAnh-infected Sf-9 cells.

摘要

杆状病毒拥有两种抗凋亡基因,即p35和凋亡抑制因子(iap)。在本研究中,我们从斜纹夜蛾核多角体病毒(SpltNPV)中克隆了一个新的IAP家族成员Splt-IAP4。对Splt-IAP4氨基酸序列进行ScanProsite分析表明,它在N端第18至61个氨基酸处含有一个杆状病毒IAP重复(BIR)基序,在其C端附近有一个锌指RING基序。根据氨基酸序列同源性,杆状病毒IAP可分为从IAP1到IAP5的五种类型,因此Splt-IAP4因含有单个BIR基序而被归类为IAP4。对已测序杆状病毒基因组中的六个IAP4进行比对表明,Splt-IAP4基因与滨海斜纹夜蛾NPV(SpliNPV)IAP4的氨基酸序列相似性为74%,高于其他杆状病毒IAP4蛋白,表明其具有进化保守性。在瞬时表达试验中,Splt-IAP4无法抑制由缺乏p35的突变型AcMNPV(vAcAnh)感染诱导的凋亡,也无法挽救Sf-9细胞中因凋亡而导致的突变病毒复制。细胞活力分析表明,Splt-IAP4不能抑制或延迟vAcAnh感染诱导的Sf-9细胞凋亡。综上所述,在vAcAnh感染的Sf-9细胞中,Splt-IAP4没有抗凋亡功能。

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