tRNAs promote nuclear import of HIV-1 intracellular reverse transcription complexes.
tRNAs promote nuclear import of HIV-1 intracellular reverse transcription complexes.
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DOI：
10.1371/journal.pbio.0040332
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发表时间：
2006-10
期刊：
影响因子：
9.8
通讯作者：
Fassati A
中科院分区：
文献类型：
--
作者：
Zaitseva L;Myers R;Fassati A
感染非分裂细胞是HIV-1的一种生物学特性，对于病毒传播和AIDS发病至关重要。这种性质取决于细胞内逆转录和前整合复合物（RTC/PIC）的核输入。为了鉴定参与HIV-1 RTCs核输入的细胞因子，通过色谱法分离胞质提取物，并通过核输入测定法检测输入活性。获得了一个接近均匀的级分，它在诱导纯化和标记的RTCs的核输入中是活跃的。活性组分中含有tRNA，大部分具有缺陷的3′ CCA末端。这种tRNA在体外合成时促进HIV-1 RTC核输入。将活性tRNA掺入病毒颗粒中并从病毒颗粒中回收。突变分析表明，反密码子环介导的病毒复合物的结合，而T-臂可能与细胞因子参与核输入。这些tRNA种类以能量和温度依赖性的方式有效地自行积累到细胞核中。含有MLV gag的HIV-1突变体没有掺入能够诱导HIV-1 RTC核输入的tRNA物质，并且未能感染细胞周期停滞的细胞。在这里，我们提供的证据表明，至少有一些tRNA种类可以输入到人类细胞的细胞核，并促进HIV-1核输入。HIV-1逆转录和整合前复合物（RTC）被输入到细胞核中，在那里它们整合到细胞DNA中;一些tRNA物质进行核再摄取并促进HIV RTC核输入。
Infection of non-dividing cells is a biological property of HIV-1 crucial for virus transmission and AIDS pathogenesis. This property depends on nuclear import of the intracellular reverse transcription and pre-integration complexes (RTCs/PICs). To identify cellular factors involved in nuclear import of HIV-1 RTCs, cytosolic extracts were fractionated by chromatography and import activity examined by the nuclear import assay. A near-homogeneous fraction was obtained, which was active in inducing nuclear import of purified and labeled RTCs. The active fraction contained tRNAs, mostly with defective 3′ CCA ends. Such tRNAs promoted HIV-1 RTC nuclear import when synthesized in vitro. Active tRNAs were incorporated into and recovered from virus particles. Mutational analyses indicated that the anticodon loop mediated binding to the viral complex whereas the T-arm may interact with cellular factors involved in nuclear import. These tRNA species efficiently accumulated into the nucleus on their own in a energy- and temperature-dependent way. An HIV-1 mutant containing MLV gag did not incorporate tRNA species capable of inducing HIV-1 RTC nuclear import and failed to infect cell cycle–arrested cells. Here we provide evidence that at least some tRNA species can be imported into the nucleus of human cells and promote HIV-1 nuclear import. HIV-1 reverse transcription and pre-integration complexes (RTC) are imported into the nucleus, where they integrate into the cellular DNA; some tRNA species undergo nuclear re-uptake and promote HIV RTC nuclear import.
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