Madiraju M V, Templin A, Clark A J
Department of Molecular Biology, University of California, Berkeley 94720.
Proc Natl Acad Sci U S A. 1988 Sep;85(18):6592-6. doi: 10.1073/pnas.85.18.6592.
A mutation partially suppressing the UV sensitivity caused by recF143 in a uvrA6 background was located at codon 37 of recA where GTG (valine) became ATG (methionine). This mutation, originally named srf-803, was renamed recA803. Little if any suppression of the recF143 defect in UV induction of a lexA regulon promoter was detected. This led to the hypothesis that a defect in recombination repair of UV damage was suppressed by recA803. The mutant RecA protein (RecA803) was purified and compared with wild-type protein (RecA+) as a catalyst of formation of joint molecules. Under suboptimal conditions, RecA803 produces both a higher rate of formation and a higher yield of joint molecules. The suboptimal conditions tested included addition of single-stranded DNA binding protein to single-stranded DNA prior to addition of RecA. We hypothesize that the ability of RecA803 to overcome interference by single-stranded DNA binding protein is the property that allows recA803 to suppress partially the deficiency in repair caused by recF mutations in the uvrA6 background. Implications of this hypothesis for the function of RecF protein in recombination are discussed.
在uvrA6背景下,一个部分抑制recF143所导致的紫外线敏感性的突变位于recA的第37密码子处,此处GTG(缬氨酸)变为ATG(甲硫氨酸)。这个最初命名为srf - 803的突变,后来被重新命名为recA803。在紫外线诱导lexA调控子启动子时,几乎没有检测到对recF143缺陷的抑制作用。这导致了一种假设,即recA803抑制了紫外线损伤的重组修复缺陷。对突变型RecA蛋白(RecA803)进行了纯化,并将其作为连接分子形成的催化剂与野生型蛋白(RecA +)进行比较。在次优条件下,RecA803产生连接分子的形成速率更高,产量也更高。所测试的次优条件包括在添加RecA之前向单链DNA中添加单链DNA结合蛋白。我们假设RecA803克服单链DNA结合蛋白干扰的能力是使得recA803能够部分抑制uvrA6背景下recF突变所导致的修复缺陷的特性。讨论了这一假设对RecF蛋白在重组中功能的影响。