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DinI和RecX调节大肠杆菌K-12中的RecA-DNA结构。

DinI and RecX modulate RecA-DNA structures in Escherichia coli K-12.

作者信息

Renzette Nicholas, Gumlaw Nathan, Sandler Steven J

机构信息

Molecular and Cell Biology Graduate Program, Morrill Science Center, University of Massachusetts at Amherst, Amherst, MA 01003, USA.

出版信息

Mol Microbiol. 2007 Jan;63(1):103-15. doi: 10.1111/j.1365-2958.2006.05496.x. Epub 2006 Dec 5.

Abstract

RecA plays a central role in recombination, DNA repair and SOS induction through forming a RecA-DNA helical filament. Biochemical observations show that at low ratios to RecA, DinI and RecX stabilize and destabilize RecA-DNA filaments, respectively, and that the C-terminal 17 residues of RecA are important for RecX function. RecA-DNA filament formation was assayed in vivo using RecA-GFP foci formation in log-phase and UV-irradiated cells. In log-phase cells, dinI mutants have fewer foci than wild type and that recX mutants have more foci than wild type. A recADelta17::gfp mutant had more foci like a recX mutant. dinI recX double mutants have the same number of foci as dinI mutants alone, suggesting that dinI is epistatic to recX. After UV treatment, the dinI, recX and dinI recX mutants differed in their ability to form foci. All three mutants had fewer foci than wild type. The dinI mutant's foci persisted longer than wild-type foci. Roles of DinI and RecX after UV treatment differed from those during log-phase growth and may reflect the different DNA substrates, population of proteins or amounts during the SOS response. These experiments give new insight into the roles of these proteins.

摘要

RecA通过形成RecA-DNA螺旋丝在重组、DNA修复和SOS诱导中发挥核心作用。生化观察表明,在与RecA的比例较低时,DinI和RecX分别稳定和破坏RecA-DNA丝,并且RecA的C末端17个残基对RecX功能很重要。使用对数期和紫外线照射细胞中RecA-GFP病灶形成在体内测定RecA-DNA丝的形成。在对数期细胞中,dinI突变体的病灶比野生型少,而recX突变体的病灶比野生型多。recADelta17::gfp突变体的病灶与recX突变体一样多。dinI recX双突变体的病灶数量与单独的dinI突变体相同,表明dinI对recX呈上位性。紫外线处理后,dinI、recX和dinI recX突变体在形成病灶的能力上有所不同。所有三个突变体的病灶都比野生型少。dinI突变体的病灶持续时间比野生型病灶长。紫外线处理后DinI和RecX的作用与对数期生长期间的作用不同,可能反映了SOS反应期间不同的DNA底物、蛋白质群体或数量。这些实验为这些蛋白质的作用提供了新的见解。

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