Centore Richard C, Sandler Steven J
Department of Microbiology, Morrill Science Center IV N203, University of Massachusetts at Amherst, Amherst, MA 01003, USA.
J Bacteriol. 2007 Apr;189(7):2915-20. doi: 10.1128/JB.01777-06. Epub 2007 Jan 26.
RecA is important for recombination, DNA repair, and SOS induction. In Escherichia coli, RecBCD, RecFOR, and RecJQ prepare DNA substrates onto which RecA binds. UvrD is a 3'-to-5' helicase that participates in methyl-directed mismatch repair and nucleotide excision repair. uvrD deletion mutants are sensitive to UV irradiation, hypermutable, and hyper-rec. In vitro, UvrD can dissociate RecA from single-stranded DNA. Other experiments suggest that UvrD removes RecA from DNA where it promotes unproductive reactions. To test if UvrD limits the number and/or the size of RecA-DNA structures in vivo, an uvrD mutation was combined with recA-gfp. This recA allele allows the number of RecA structures and the amount of RecA at these structures to be assayed in living cells. uvrD mutants show a threefold increase in the number of RecA-GFP foci, and these foci are, on average, nearly twofold higher in relative intensity. The increased number of RecA-green fluorescent protein foci in the uvrD mutant is dependent on recF, recO, recR, recJ, and recQ. The increase in average relative intensity is dependent on recO and recQ. These data support an in vivo role for UvrD in removing RecA from the DNA.
RecA对于重组、DNA修复和SOS诱导至关重要。在大肠杆菌中,RecBCD、RecFOR和RecJQ会准备好RecA结合的DNA底物。UvrD是一种3'至5'解旋酶,参与甲基导向的错配修复和核苷酸切除修复。uvrD缺失突变体对紫外线照射敏感,具有高突变性且重组频率高。在体外,UvrD可以使RecA从单链DNA上解离。其他实验表明,UvrD会将RecA从促进非生产性反应的DNA上移除。为了测试UvrD在体内是否限制RecA-DNA结构的数量和/或大小,将uvrD突变与recA-gfp结合。这个recA等位基因使得可以在活细胞中检测RecA结构的数量以及这些结构上RecA的量。uvrD突变体显示RecA-GFP焦点数量增加了三倍,并且这些焦点的平均相对强度几乎高出近两倍。uvrD突变体中RecA-绿色荧光蛋白焦点数量的增加依赖于recF、recO、recR、recJ和recQ。平均相对强度的增加依赖于recO和recQ。这些数据支持UvrD在体内从DNA上移除RecA的作用。