Inhibition of HIV-1 infection by TNPO3 depletion is determined by capsid and detectable after viral cDNA enters the nucleus.
Inhibition of HIV-1 infection by TNPO3 depletion is determined by capsid and detectable after viral cDNA enters the nucleus.
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DOI：
10.1186/1742-4690-8-98
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发表时间：
2011-12-06
期刊：
影响因子：
3.3
通讯作者：
Luban J
中科院分区：
文献类型：
--
作者：
De Iaco A;Luban J
HIV-1感染非分裂细胞。这意味着病毒在整合到染色体DNA之前会穿过核孔。最近的研究表明，TnPO3是HIV-1完全传染性所必需的。TnPO3是一种核粘附素，这一事实表明它通过直接促进HIV-1的核进入而发挥作用。一些研究支持这一假设，而另一些研究则未能做到这一点。此外，一些研究表明，TNPO3通过HIV-1整合酶(IN)发挥作用，另一些研究表明，它通过衣壳(CA)发挥作用。为了阐明TNPO3促进HIV-1感染的机制，我们设计了一组27个单循环HIV-1载体，每个载体携带不同的CA突变，并鉴定了它们转导TNPO3被击倒的细胞的能力(KD)。与野生型(WT)HIV-1相比，14个CA突变体相对不依赖于TNPO3。两个突变体比WT对TNPO3的依赖性更强，11个突变体实际上被TNPO3抑制。然后对野生型HIV-1和选择的三个CA突变体的病毒DNA、2-LTR环和前病毒DNA的合成效率进行了评估。对照包括用非靶向编码序列抢救TNPO3KD，RT和IN突变病毒，以及RT和IN的药物抑制剂。TNPO3KD阻断了野生型HIV-1对前病毒DNA的转导和建立，但对2-LTR环水平无明显影响。通过使用两种不同的方法(慢病毒载体和siRNA寡核苷酸转染法)获得TNPO3 KD，通过攻击三种不同的细胞类型，通过使用两种不同的挑战病毒，每种病毒需要不同的一组PCR引物，以及通过使用VSV G或使用HIV-1 Env进行伪分型病毒，证实了PCR结果。在病毒生命周期中的一个阶段，在前整合复合体到达细胞核后可以检测到的阶段，TNPO3促进了HIV-1的传染性，而CA是依赖TNPO3的病毒决定因素。
HIV-1 infects non-dividing cells. This implies that the virus traverses the nuclear pore before it integrates into chromosomal DNA. Recent studies demonstrated that TNPO3 is required for full infectivity of HIV-1. The fact that TNPO3 is a karyopherin suggests that it acts by directly promoting nuclear entry of HIV-1. Some studies support this hypothesis, while others have failed to do so. Additionally, some studies suggest that TNPO3 acts via HIV-1 Integrase (IN), and others indicate that it acts via capsid (CA). To shed light on the mechanism by which TNPO3 contributes to HIV-1 infection we engineered a panel of twenty-seven single-cycle HIV-1 vectors each bearing a different CA mutation and characterized them for the ability to transduce cells in which TNPO3 had been knocked down (KD). Fourteen CA mutants were relatively TNPO3-independent, as compared to wild-type (WT) HIV-1. Two mutants were more TNPO3-dependent than the WT, and eleven mutants were actually inhibited by TNPO3. The efficiency of the synthesis of viral cDNA, 2-LTR circles, and proviral DNA was then assessed for WT HIV-1 and three select CA mutants. Controls included rescue of TNPO3 KD with non-targetable coding sequence, RT- and IN- mutant viruses, and pharmacologic inhibitors of RT and IN. TNPO3 KD blocked transduction and establishment of proviral DNA by wild-type HIV-1 with no significant effect on the level of 2-LTR circles. PCR results were confirmed by achieving TNPO3 KD using two different methodologies (lentiviral vector and siRNA oligonucleotide transfection); by challenging three different cell types; by using two different challenge viruses, each necessitating different sets of PCR primers; and by pseudotyping virus with VSV G or using HIV-1 Env. TNPO3 promotes HIV-1 infectivity at a step in the virus life cycle that is detectable after the preintegration complex arrives in the nucleus and CA is the viral determinant for TNPO3 dependence.
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DOI：
10.1073/pnas.86.8.2525
发表时间：
1989-04-01
影响因子：
11.1
作者：
BROWN, PO;BOWERMAN, B;BISHOP, JM
通讯作者：
BISHOP, JM
影响因子：
6.7
作者：
Blair WS;Pickford C;Irving SL;Brown DG;Anderson M;Bazin R;Cao J;Ciaramella G;Isaacson J;Jackson L;Hunt R;Kjerrstrom A;Nieman JA;Patick AK;Perros M;Scott AD;Whitby K;Wu H;Butler SL
通讯作者：
Butler SL
影响因子：
5.4
作者：
Brussel, A;Sonigo, P
通讯作者：
Sonigo, P
影响因子：
5.4
作者：
Forshey, BM;Shi, J;Aiken, C
通讯作者：
Aiken, C
影响因子：
30.3
作者：
Lee K;Ambrose Z;Martin TD;Oztop I;Mulky A;Julias JG;Vandegraaff N;Baumann JG;Wang R;Yuen W;Takemura T;Shelton K;Taniuchi I;Li Y;Sodroski J;Littman DR;Coffin JM;Hughes SH;Unutmaz D;Engelman A;KewalRamani VN
通讯作者：
KewalRamani VN