Intracytoplasmic maturation of the human immunodeficiency virus type 1 reverse transcription complexes determines their capacity to integrate into chromatin.
Intracytoplasmic maturation of the human immunodeficiency virus type 1 reverse transcription complexes determines their capacity to integrate into chromatin.
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DOI：
10.1186/1742-4690-3-4
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发表时间：
2006-01-12
期刊：
影响因子：
3.3
通讯作者：
Bukrinsky M
中科院分区：
文献类型：
--
作者：
Iordanskiy S;Berro R;Altieri M;Kashanchi F;Bukrinsky M
HIV-1生命周期的早期事件包括病毒核心进入靶细胞、组装执行逆转录的逆转录复合物(RTC)、其转化为具有整合能力的复合物(称为预整合复合物(PIC))、复合物运输到细胞核中，以及最后将病毒DNA整合到染色质中。这些过程的分子细节和时间组织仍然是艾滋病毒生物学中研究最少和最具争议的问题之一。为了定量评估 HIV-1 RTC 的成熟和核转位，通过实时 PCR 分析了从感染 MLV Env 假型 HIV-1 的 HeLa 细胞的细胞核 (nRTC) 和细胞质 (cRTC) 中分离的核蛋白复合物。虽然大多数复合物在细胞质中完成逆转录，但有些复合物在完成 DNA 合成之前进入细胞核。当逆转录酶抑制剂阻断逆转录时，HIV特异性RNA复合物可以进入细胞核，尽管RNA复合物的入核效率低于含有DNA的RTC。对细胞周期 G2/M 期感染的同步细胞中 RTC 核输入的分析表明，与非同步细胞相比，含有不完整 HIV-1 DNA 的 RTC 细胞核富集，其中具有完整逆转录本的 RTC 占主导地位。免疫沉淀分析鉴定出与 cRTC 和 nRTC 相关的病毒蛋白 IN、Vpr、MA 以及细胞 Ini1 和 PML，而 CA 仅在 cRTC 中检测到，而 RT 在 nRTC 中减少。复合物的细胞质成熟与抗Vpr和抗IN抗体的免疫反应性增加以及与RT抗体的反应性降低相关。 cRTCs和nRTCs均在体外进行内源逆转录反应。与 cRTC 相比，nRTC 的体外逆转录完成并没有增加它们与染色质的整合。这些结果表明 RTC 成熟主要发生在细胞质中。含有 RT 和不完整 DNA 的未成熟 RTC 可以在有丝分裂和完成逆转录过程中易位到细胞核中，但整合有缺陷。
The early events of the HIV-1 life cycle include entry of the viral core into target cell, assembly of the reverse transcription complex (RTCs) performing reverse transcription, its transformation into integration-competent complexes called pre-integration complexes (PICs), trafficking of complexes into the nucleus, and finally integration of the viral DNA into chromatin. Molecular details and temporal organization of these processes remain among the least investigated and most controversial problems in the biology of HIV. To quantitatively evaluate maturation and nuclear translocation of the HIV-1 RTCs, nucleoprotein complexes isolated from the nucleus (nRTC) and cytoplasm (cRTC) of HeLa cells infected with MLV Env-pseudotyped HIV-1 were analyzed by real-time PCR. While most complexes completed reverse transcription in the cytoplasm, some got into the nucleus before completing DNA synthesis. The HIV-specific RNA complexes could get into the nucleus when reverse transcription was blocked by reverse transcriptase inhibitor, although nuclear import of RNA complexes was less efficient than of DNA-containing RTCs. Analysis of the RTC nuclear import in synchronized cells infected in the G2/M phase of the cell cycle showed enrichment in the nuclei of RTCs containing incomplete HIV-1 DNA compared to non-synchronized cells, where RTCs with complete reverse transcripts prevailed. Immunoprecipitation assays identified viral proteins IN, Vpr, MA, and cellular Ini1 and PML associated with both cRTCs and nRTCs, whereas CA was detected only in cRTCs and RT was diminished in nRTCs. Cytoplasmic maturation of the complexes was associated with increased immunoreactivity with anti-Vpr and anti-IN antibodies, and decreased reactivity with antibodies to RT. Both cRTCs and nRTCs carried out endogenous reverse transcription reaction in vitro. In contrast to cRTCs, in vitro completion of reverse transcription in nRTCs did not increase their integration into chromatin. These results suggest that RTC maturation occurs predominantly in the cytoplasm. Immature RTCs containing RT and incomplete DNA can translocate into the nucleus during mitosis and complete reverse transcription, but are defective for integration.
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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：
Fassati, A;Goff, SP
通讯作者：
Goff, SP
DOI：
10.1073/pnas.90.13.6125
发表时间：
1993-07-01
影响因子：
11.1
作者：
BUKRINSKY, MI;SHAROVA, N;STEVENSON, M
通讯作者：
STEVENSON, M
影响因子：
5.4
作者：
FARNET, CM;HASELTINE, WA
通讯作者：
HASELTINE, WA
DOI：
10.1073/pnas.87.11.4164
发表时间：
1990-06-01
影响因子：
11.1
作者：
FARNET, CM;HASELTINE, WA
通讯作者：
HASELTINE, WA