Study of anti-APOBEC3G activities of HIV-1 Vif proteins among different subtypes

HIV-1 Vif蛋白不同亚型抗APOBEC3G活性研究

• 批准号: 18590460
• 负责人: TOKUNAGA Kenzo
• 金额: $ 2.57万
• 依托单位: National Institute of Infectious Diseases
• 依托单位国家: 日本
• 项目类别: Grant-in-Aid for Scientific Research (C)
• 财政年份: 2006
• 资助国家: 日本
• 起止时间: 2006 至 2007
• 项目状态: 已结题
• 来源: https://kaken.nii.ac.jp/grant/KAKENHI-PROJECT-18590460/
• 关键词: HIV-1; subtype; Vif; APOBEC3

Antiretroviral cytidine deaminase APOBEC3G, which is abundantly expressed in peripheral blood lymphocytes and macrophages, strongly protects these cells against HIV-I infection. The Vif protein of HIV-1 overcomes this antiviral effect by inducing proteasome-mediated degradation of APOBEC3G, and is a key for maintaining viral infectivity. The vif gene, which is 579-bp long, displays high genetic diversity among HIV-I subtypes and it is therefore intriguing to address whether Vif proteins derived from different subtypes might be differ in the viral defense activity against APOBEC3G. To test this, we created expression plasmids encoding Vif proteins derived from subtypes A, B, C, CRF01_AE, and CRF02_AG clinical isolates and compared their anti-APOBEC3G activities. Viruses, produced from cells in the presence of APOBEC3G and Vif proteins of different subtypes, showed differential viral infectivities, that is, subtype C-derived Vif proteins showed exclusively high anti-APOBEC3G activities, compared with any. other subtype-derived Vif proteins, and this was found to be because of the different levels of proteasomal APOBEC3G degradation by Vif, depending on subtypes. To determine which portion of subtype C-Vif protein would be responsible for the robust anit-APOBEC3G activities, we created chimeric Vif constructs between subtypes B and C, and further pursued the responsible amino acid (s) by generating point mutants of Vif plasmids. As a result, we identified two amino acids at N-terminus responsible for subtype C-Vif-specific anti-APOBEC3G activity. Intriguingly, one of these amino acids is contained in the conserved sequence which is proposed to be involved in the interaction with APOBEC3G, suggesting that subtype C-vif might be able to bind APOBEC3G more efficiently than do Vif proteins of other subtypes. These results imply that biological differences of Vif proteins among subtypes might have an impact on viral transmissibility.

抗逆转录病毒胞苷脱氨酶APOBEC 3G在外周血淋巴细胞和巨噬细胞中大量表达，强烈保护这些细胞免受HIV-I感染。HIV-1的Vif蛋白通过诱导蛋白酶体介导的APOBEC 3G降解来克服这种抗病毒作用，并且是维持病毒感染性的关键。长579 bp的vif基因在HIV-I亚型中显示出高度的遗传多样性，因此解决来自不同亚型的Vif蛋白是否在针对APOBEC 3G的病毒防御活性上可能不同是有趣的。为了测试这一点，我们创建了编码来自亚型A、B、C、CRF 01_AE和CRF 02_AG临床分离株的Vif蛋白的表达质粒，并比较了它们的抗APOBEC 3G活性。在不同亚型的APOBEC 3G和Vif蛋白存在下从细胞产生的病毒显示出不同的病毒感染性，也就是说，亚型C衍生的Vif蛋白与任何病毒相比仅显示出高的抗APOBEC 3G活性。其他亚型衍生的Vif蛋白，发现这是因为Vif对蛋白酶体APOBEC 3G的降解水平不同，这取决于亚型。为了确定亚型C-Vif蛋白的哪一部分将负责稳健的抗APOBEC 3G活性，我们在亚型B和C之间创建嵌合Vif构建体，并通过产生Vif质粒的点突变体进一步追踪负责的氨基酸。因此，我们在N-末端鉴定了两个负责亚型C-Vif特异性抗APOBEC 3G活性的氨基酸。有趣的是，这些氨基酸之一包含在保守序列中，该保守序列被认为参与与APOBEC 3G的相互作用，这表明亚型C-vif可能能够比其他亚型的Vif蛋白更有效地结合APOBEC 3G。这些结果表明，不同亚型之间Vif蛋白的生物学差异可能对病毒的传播性产生影响。

项目成果

APOBEC3ファミリーによるLINE-1レトロトランスポゾン抑制効果.

APOBEC3 家族对 LINE-1 逆转录转座子的抑制作用。

• 发表时间: 2006
• 作者: 徳永研三;木ノ本正信;志村まり;石坂幸人;佐多徹太郎
• 通讯作者: 佐多徹太郎

All APOBEC3 family proteins differentially inhibit LINE-1 retrotransposition.

• DOI: 10.1093/nar/gkm181
• 发表时间: 2007
• 期刊: NUCLEIC ACIDS RESEARCH
• 影响因子: 14.9
• 作者: Kinomoto, Masanobu;Kanno, Takayuki;Shimura, Mari;Ishizaka, Yukihito;Kojima, Asato;Kurata, Takeshi;Sata, Tetsutaro;Tokunaga, Kenzo
• 通讯作者: Tokunaga, Kenzo

Differential inhibitory activities of APOBEC3 family proteins on retroviruses and LINE-1 retrotransposon.

APOBEC3 家族蛋白对逆转录病毒和 LINE-1 逆转录转座子的差异抑制活性。

• 发表时间: 2006
• 作者: Kinomoto M;Shimura M;Ishizaka Y;Sata T;and Tokunaga K
• 通讯作者: and Tokunaga K

HIV-1 Vpr induces DNA double-strand breaks.

• DOI: 10.1158/0008-5472.can-05-3144
• 发表时间: 2006-01
• 期刊: Cancer research
• 影响因子: 11.2
• 作者: H. Tachiwana;M. Shimura;Chikako Nakai-Murakami;K. Tokunaga;Y. Takizawa;T. Sata;H. Kurumizaka;Y. Ish

HIV-1 Vprによる核膜異常.

HIV-1 Vpr 引起的核膜异常。

• 发表时间: 2007
• 作者: 志村まり;前島一博;宮澤雅之;森美樹;徳永研三;佐多徹太郎;今本尚子;瀧澤俊博;石坂幸人
• 通讯作者: 石坂幸人