A NUCLEOPROTEIN COMPLEX MEDIATES THE INTEGRATION OF RETROVIRAL DNA
A NUCLEOPROTEIN COMPLEX MEDIATES THE INTEGRATION OF RETROVIRAL DNA
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DOI：
10.1101/gad.3.4.469
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发表时间：
1989-04-01
影响因子：
10.5
通讯作者：
VARMUS, HE
中科院分区：
文献类型：
--
作者：
BOWERMAN, B;BROWN, PO;VARMUS, HE
病毒DNA整合到宿主基因组中是逆转录病毒生命周期中必不可少的一步。为了更好地了解这一过程，我们使用病毒DNA的物理分析和整合活性的功能分析，研究了急性感染小鼠白血病病毒(MLV)的细胞中病毒DNA的自然状态(Brown等人)。1987)。病毒DNA和整合活性在速度沉降、凝胶过滤和密度平衡离心过程中相互作用，表明病毒DNA存在于一个大的(约160S)核蛋白复合体中，它包括体外整合活性所需的所有功能。免疫沉淀法分析表明，病毒衣壳蛋白是活性核蛋白复合体的一部分，但只有一小部分抗衣壳血清识别该复合体意味着该蛋白受到构象限制。这种结构中的病毒DNA可以被核酸酶访问；核酸酶对复合体完整性的影响表明，整合能力强的颗粒来自细胞外病毒粒子的核心，并与其相似。
The intergration of viral DNA into the host genome is an essential step in the retrovirus life cycle. To understand this process better, we have examined the native state of viral DNA in cells acutely infected by murine leukemia virus (MLV), using both a physical assay of viral DNA and a functional assay for integration activity (Brown et al. 1987). The viral DNA and integration activity copurify during velocity sedimentation, gel filtration, and density equilibrium centrifugation, indicating that virual DNA is in a large (.apprx.160S) nucleoprotein complex that includes all functions required for integration activity in vitro. Analysis by immunoprecipitation shows that the viral capsid protein is part of the active nucleoprotein complex, but recognition of the complex by only a subset of anti-capsid sera implies that the protein is constrained conformationally. The viral DNA within this structures is accessible to nucleases; the effects of nucleases on the integrity of the complex suggest that the integration-competent particle is derived from and similar to the core of extracellular virions.