Cyclophilin A regulates HIV-1 infectivity, as demonstrated by gene targeting in human T cells
Cyclophilin A regulates HIV-1 infectivity, as demonstrated by gene targeting in human T cells
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DOI：
10.1093/emboj/20.6.1300
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发表时间：
2001-03-15
期刊：
影响因子：
11.4
通讯作者：
Luban, J
中科院分区：
文献类型：
--
作者：
Braaten, D;Luban, J
人类免疫缺陷病毒1型（HIV-1）Gag多聚蛋白结合肽基脯氨酰异构酶亲环蛋白家族的大多数成员。在已知的15种人类亲环蛋白中，亲环蛋白A（CypA）由于在HIV-1病毒粒子中被检测到而成为研究的焦点。为了确定CypA是否促进HIV-1复制，我们通过同源重组删除了人CD 4（+）T细胞中编码CypA（PPIA）的基因，PPIA（-/-）细胞中的HIV-1复制减少，并且不被环孢菌素或gag突变进一步抑制，这些突变破坏了Gag与亲环素的相互作用，表明没有其他亲环素家族成员促进HIV-1复制。缺陷复制表型对野生型HIV-1是特异性的，因为HIV-2/SIV分离株以及携带gag突变的HIV-1赋予环孢菌素抗性，在PPIA（+/+）和PPIA（-/-）细胞中复制相同。CypA在PPIA（-/-）细胞中的稳定再表达使HIV-1复制恢复到与CypA稳态水平相关的程度。最后，来自PPIA（-/-）细胞的病毒粒子没有明显的生化异常，但比来自轻度型Tells的病毒粒子感染性低。这些数据正式证明CypA调节HIV-1病毒粒子的感染性。
The human immunodeficiency virus type 1 (HIV-1) Gag polyprotein binds most members of the cyclophilin family of peptidyl-prolyl isomerases. Of 15 known human cyclophilins, cyclophilin A (CypA) has been the focus of investigation because it was detected in HIV-1 virions. To determine whether CypA promotes HIV-1 replication, we deleted the gene encoding CypA (PPIA) in human CD4(+) T cells by homologous recombination, HIV-1 replication in PPIA(-/-) cells was decreased and not inhibited further by cyclosporin or gag mutations that disrupt Gag's interaction with cyclophilins, indicating that no other cyclophilin family members promote HIV-1 replication. The defective replication phenotype was specific for wild-type HIV-1 since HIV-2/SIV isolates, as well as HIV-1 bearing a gag mutation that confers cyclosporin resistance, replicated the same in PPIA(+/+) and PPIA(-/-) cells. Stable re-expression of CypA in PPIA(-/-) cells restored HIV-1 replication to an extent that correlated with steady-state levels of CypA, Finally, virions from PPIA(-/-) cells possessed no obvious biochemical abnormalities but were less infectious than virions from mild-type tells. These data formally demonstrate that CypA regulates the infectivity of HIV-1 virions.