Farnet C M, Bushman F D
Salk Institute for Biological Studies, La Jolla, California 92024, USA.
Cell. 1997 Feb 21;88(4):483-92. doi: 10.1016/s0092-8674(00)81888-7.
We present data indicating that a host protein is important for function of HIV-1 preintegration complexes (PICs) in vitro. PICs partially purified from infected cells were subjected to gel filtration in 600 mM KCl, which removed a factor required for integration without fully disrupting PICs. Addition of an extract from uninfected cells restored activity. Fractionation of the complementing activity yielded HMG I(Y), a nonhistone chromosomal protein important for transcriptional control and chromosomal architecture. Complementing activity could be isolated from PICs, and activity could be depleted from such fractions with an antibody against HMG I(Y). Recombinant HMG I(Y) also complemented salt-stripped complexes. The finding that a host protein is required for integration by HIV PICs parallels findings in several well-studied transposition and site-specific recombination systems.
我们提供的数据表明，一种宿主蛋白对于HIV-1整合前复合物（PICs）在体外的功能很重要。从感染细胞中部分纯化得到的PICs在600 mM KCl中进行凝胶过滤，这去除了整合所需的一个因子，但并未完全破坏PICs。添加未感染细胞的提取物可恢复活性。对互补活性进行分级分离得到了HMG I(Y)，这是一种对转录调控和染色体结构很重要的非组蛋白染色体蛋白。互补活性可从PICs中分离出来，并且这种级分中的活性可用抗HMG I(Y)抗体去除。重组HMG I(Y)也能互补经盐处理去除某些成分的复合物。HIV PICs整合需要宿主蛋白这一发现与几个经过充分研究的转座和位点特异性重组系统中的发现相似。
