Functional coupling between HIV-1 integrase and the SWI/SNF chromatin remodeling complex for efficient in vitro integration into stable nucleosomes.
Functional coupling between HIV-1 integrase and the SWI/SNF chromatin remodeling complex for efficient in vitro integration into stable nucleosomes.
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DOI：
10.1371/journal.ppat.1001280
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发表时间：
2011-02-10
期刊：
影响因子：
6.7
通讯作者：
Parissi V
中科院分区：
文献类型：
--
作者：
Lesbats P;Botbol Y;Chevereau G;Vaillant C;Calmels C;Arneodo A;Andreola ML;Lavigne M;Parissi V
稳定的HIV-1感染的建立需要逆转录病毒基因组有效整合到宿主DNA中。染色质结构控制这一过程的分子机制尚未阐明。我们在这里表明，稳定相关的核小体强烈抑制在体外两个病毒末端的整合，通过降低DNA整合酶的可及性。通过SWI/SNF复合物重塑染色质化模板，其INI1主要组分与IN相互作用，恢复并重定向到稳定核小体区域的全位点整合。在通过其他人类重塑因子（如SNF2H或缺乏整合酶结合蛋白INI1的BRG 1）重塑后未观察到这些作用。这表明修复过程取决于IN和整个SWI/SNF复合体之间的直接相互作用，支持重塑和整合复合体之间的功能耦合。此外，从文献中选择的超过40，000个非冗余细胞整合位点与核小体占据预测之间的计算机比较也支持HIV-1整合在感染细胞中内在核小体密度较弱的基因组区域中被促进。我们的数据表明，一些染色质结构可以是难治性的整合和核小体重塑和HIV-1整合之间的耦合需要克服这一自然障碍。HIV-1的感染和致病性需要其基因组的DNA拷贝整合到宿主细胞染色体中。这导致宿主和逆转录病毒之间的稳定关联，防止其从患者中完全根除和耐药病毒的持续存档。即使病毒整合酶催化的反应机制现在是众所周知的，它与宿主染色质的相互作用是不完全理解。染色质是高度结构化的，这是由于核小体中的DNA压缩，其中DNA可及性可以影响整合的效率和选择性。使用体外试验，使我们能够复制和监测容易地整合到一个受体染色质化的质粒，我们表明，一个稳定的和有组织的核小体的存在下，防止病毒DNA的整合。此外，我们报告说，整合到核小体区域可以恢复，如果它是与细胞因子重塑染色质结构。我们的研究结果表明，HIV-1整合复合物和染色质维持宿主机制之间存在强有力的功能性相互作用，这是HIV-1基因组有效整合到细胞DNA中所必需的。这表明抑制HIV-1复制的潜在新的治疗靶点，以及在基因治疗中调节慢病毒载体介导的整合的选择性的潜在新方法。
Establishment of stable HIV-1 infection requires the efficient integration of the retroviral genome into the host DNA. The molecular mechanism underlying the control of this process by the chromatin structure has not yet been elucidated. We show here that stably associated nucleosomes strongly inhibit in vitro two viral-end integration by decreasing the accessibility of DNA to integrase. Remodeling of the chromatinized template by the SWI/SNF complex, whose INI1 major component interacts with IN, restores and redirects the full-site integration into the stable nucleosome region. These effects are not observed after remodeling by other human remodeling factors such as SNF2H or BRG1 lacking the integrase binding protein INI1. This suggests that the restoration process depends on the direct interaction between IN and the whole SWI/SNF complex, supporting a functional coupling between the remodeling and integration complexes. Furthermore, in silico comparison between more than 40,000 non-redundant cellular integration sites selected from literature and nucleosome occupancy predictions also supports that HIV-1 integration is promoted in the genomic region of weaker intrinsic nucleosome density in the infected cell. Our data indicate that some chromatin structures can be refractory for integration and that coupling between nucleosome remodeling and HIV-1 integration is required to overcome this natural barrier. The infection and pathogenicity of HIV-1 requires the integration of a DNA copy of its genome into host cell chromosomes. This leads to a stable association between the host and the retrovirus, preventing its total eradication from the patient and the constant archiving of drug-resistant viruses. Even if the reaction mechanism catalyzed by the viral integrase is now well known, its interaction with the host chromatin is incompletely understood. Chromatin is highly structured due to DNA compaction in nucleosomes where the DNA accessibility can affect both the efficiency and the selectivity of integration. Using an in vitro assay allowing us to reproduce and monitor easily the integration into an acceptor chromatinized plasmid, we show that the presence of a stable and organized nucleosome prevents the integration of the viral DNA. Additionally, we report that the integration into nucleosome regions can be restored if it is coupled with cellular factors that remodel the chromatin structure. Our results indicate a strong functional interaction between the HIV-1 integration complex and the chromatin maintenance host machinery required for efficient integration of the HIV-1 genome into the cellular DNA. This suggests potentially new therapeutical targets for inhibiting HIV-1 replication, and also potentially new ways for modulating the selectivity of the lentiviral vector-mediated integration in gene therapy.
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