RecF蛋白与RecO、RecR及单链DNA结合蛋白的相互作用揭示了RecF-RecO-RecR复合物在DNA修复和重组中的作用。
Interactions of RecF protein with RecO, RecR, and single-stranded DNA binding proteins reveal roles for the RecF-RecO-RecR complex in DNA repair and recombination.
作者信息
Hegde S P, Qin M H, Li X H, Atkinson M A, Clark A J, Rajagopalan M, Madiraju M V
机构信息
Department of Microbiology, University of Texas Health Center at Tyler 75710, USA.
出版信息
Proc Natl Acad Sci U S A. 1996 Dec 10;93(25):14468-73. doi: 10.1073/pnas.93.25.14468.
The products of the recF, recO, and recR genes are thought to interact and assist RecA in the utilization of single-stranded DNA precomplexed with single-stranded DNA binding protein (Ssb) during synapsis. Using immunoprecipitation, size-exclusion chromatography, and Ssb protein affinity chromatography in the absence of any nucleotide cofactors, we have obtained the following results: (i) RecF interacts with RecO, (ii) RecF interacts with RecR in the presence of RecO to form a complex consisting of RecF, RecO, and RecR (RecF-RecO-RecR); (iii) RecF interacts with Ssb protein in the presence of RecO. These data suggested that RecO mediates the interactions of RecF protein with RecR and with Ssb proteins. Incubation of RecF, RecO, RecR, and Ssb proteins resulted in the formation of RecF-RecO-Ssb complexes; i.e., RecR was excluded. Preincubation of RecF, RecO, and RecR proteins prior to addition of Ssb protein resulted in the formation of complexes consisting of RecF, RecO, RecR, and Ssb proteins. These data suggest that one role of RecF is to stabilize the interaction of RecR with RecO in the presence of Ssb protein. Finally, we found that interactions of RecF with RecO are lost in the presence of ATP. We discuss these results to explain how the RecF-RecO-RecR complex functions as an anti-Ssb factor.
recF、recO和recR基因的产物被认为在联会过程中相互作用，并协助RecA利用与单链DNA结合蛋白（Ssb）预复合的单链DNA。在不存在任何核苷酸辅因子的情况下，我们使用免疫沉淀、尺寸排阻色谱和Ssb蛋白亲和色谱法，得到了以下结果：（i）RecF与RecO相互作用；（ii）在RecO存在的情况下，RecF与RecR相互作用形成由RecF、RecO和RecR组成的复合物（RecF-RecO-RecR）；（iii）在RecO存在的情况下，RecF与Ssb蛋白相互作用。这些数据表明，RecO介导了RecF蛋白与RecR以及与Ssb蛋白的相互作用。RecF、RecO、RecR和Ssb蛋白一起温育会形成RecF-RecO-Ssb复合物，即RecR被排除在外。在添加Ssb蛋白之前，先将RecF、RecO和RecR蛋白预温育，会形成由RecF、RecO、RecR和Ssb蛋白组成的复合物。这些数据表明，RecF的一个作用是在Ssb蛋白存在的情况下稳定RecR与RecO的相互作用。最后，我们发现，在ATP存在的情况下，RecF与RecO的相互作用消失。我们讨论这些结果，以解释RecF-RecO-RecR复合物如何作为一种抗Ssb因子发挥作用。
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