Masson J Y, Stasiak A Z, Stasiak A, Benson F E, West S C
Imperial Cancer Research Fund, Clare Hall Laboratories, South Mimms, Hertfordshire EN6 3LD, United Kingdom.
Proc Natl Acad Sci U S A. 2001 Jul 17;98(15):8440-6. doi: 10.1073/pnas.111005698.
In vertebrates, the RAD51 protein is required for genetic recombination, DNA repair, and cellular proliferation. Five paralogs of RAD51, known as RAD51B, RAD51C, RAD51D, XRCC2, and XRCC3, have been identified and also shown to be required for recombination and genome stability. At the present time, however, very little is known about their biochemical properties or precise biological functions. As a first step toward understanding the roles of the RAD51 paralogs in recombination, the human RAD51C and XRCC3 proteins were overexpressed and purified from baculovirus-infected insect cells. The two proteins copurify as a complex, a property that reflects their endogenous association observed in HeLa cells. Purified RAD51C--XRCC3 complex binds single-stranded, but not duplex DNA, to form protein--DNA networks that have been visualized by electron microscopy.
在脊椎动物中,RAD51蛋白对于基因重组、DNA修复和细胞增殖是必需的。已鉴定出RAD51的五个旁系同源物,即RAD51B、RAD51C、RAD51D、XRCC2和XRCC3,它们也被证明是重组和基因组稳定性所必需的。然而,目前对它们的生化特性或精确生物学功能知之甚少。作为了解RAD51旁系同源物在重组中作用的第一步,人RAD51C和XRCC3蛋白在杆状病毒感染的昆虫细胞中过表达并纯化。这两种蛋白作为复合物共同纯化,这一特性反映了它们在HeLa细胞中观察到的内源性结合。纯化的RAD51C-XRCC3复合物结合单链而非双链DNA,形成已通过电子显微镜观察到的蛋白质-DNA网络。