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recX,一种与大肠杆菌中的recA基因共转录的新SOS基因。

recX, a new SOS gene that is co-transcribed with the recA gene in Escherichia coli.

作者信息

Pagès Vincent, Koffel-Schwartz Nicole, Fuchs Robert P P

机构信息

UPR9003, CNRS Cancérogenèse et Mutagenèse Moléculaire et Structurale, ESBS, Blvd Sebastien Brant, 67400, Strasbourg, France.

出版信息

DNA Repair (Amst). 2003 Mar 1;2(3):273-84. doi: 10.1016/s1568-7864(02)00217-3.

Abstract

recX is a small open reading frame located downstream of recA that is conserved in many bacteria. In Escherichia coli, the recX gene (also named oraA) is a 501 bp open reading frame that encodes a predicted basic protein. Transcriptional analysis by Northern blots showed that in E. coli the recX gene is SOS-regulated. Primer extension data and transcriptional fusions indicate that recX transcription is down regulated with respect to recA by an intrinsic transcription terminator that is located between the recA and recX coding sequences. Despite the presence of this terminator, a recA-recX message resulting from transcriptional readthrough is detected at a level of 5-10% of the recA message. In addition, transcriptional/translational fusion experiments show that recX expression is further down regulated at the translational level reaching an estimated protein level about 500-fold lower than RecA. Strains in which the recX gene was disrupted were constructed by insertion of an antibiotic resistance cassette. The survival after UV irradiation, the spontaneous and UV-induced mutation rates were not significantly different in these recX strains compared to the corresponding wild type strain. Overexpression of RecA was shown to be lethal in a recX deletion strain in Pseudomonas aeruginosa [J. Bacteriol. 175 (1993) 2451], Mycobacterium tuberculosis [Mol. Microbiol. 30 (1998) 525] and Streptomyces lividans [J. Bacteriol. 182 (2000) 4005] suggesting that the recX gene may act as a regulator of recA. In contrast in E. coli, in a recX deletion strain, RecA overexpression is neither toxic nor is the expression of the recA(+) gene affected in a recX deletion strain at the basal level or after UV induction.

摘要

recX是一个位于recA下游的小开放阅读框,在许多细菌中保守存在。在大肠杆菌中,recX基因(也称为oraA)是一个501 bp的开放阅读框,编码一种预测的碱性蛋白。Northern印迹法进行的转录分析表明,在大肠杆菌中recX基因受SOS调控。引物延伸数据和转录融合表明,recX转录相对于recA通过位于recA和recX编码序列之间的一个内在转录终止子而下调。尽管存在这个终止子,但转录通读产生的recA-recX信使RNA在recA信使RNA水平的5%-10%被检测到。此外,转录/翻译融合实验表明,recX表达在翻译水平进一步下调,估计蛋白质水平比RecA低约500倍。通过插入抗生素抗性盒构建了recX基因被破坏的菌株。与相应的野生型菌株相比,这些recX菌株在紫外线照射后的存活率、自发和紫外线诱导的突变率没有显著差异。在铜绿假单胞菌[《细菌学杂志》175 (1993) 2451]、结核分枝杆菌[《分子微生物学》30 (1998) 525]和变铅青链霉菌[《细菌学杂志》182 (2000) 4005]中,RecA的过表达在recX缺失菌株中被证明是致死的,这表明recX基因可能作为recA的调节因子。相比之下,在大肠杆菌中,在recX缺失菌株中,RecA过表达既无毒,recA(+)基因在基础水平或紫外线诱导后在recX缺失菌株中的表达也不受影响。

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