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耐辐射球菌中LexA阻遏蛋白结合位点的保守性。

Conservation of the LexA repressor binding site in Deinococcus radiodurans.

作者信息

Khan Feroz, Singh S P, Mishra B N

机构信息

Department of Biotechnology, Institute of Engineering & Technology, U. P. Technical University, Lucknow, India.

出版信息

J Integr Bioinform. 2008 Jan 24;5(1):86. doi: 10.2390/biecoll-jib-2008-86.

Abstract

The LexA protein is a transcriptional repressor of the bacterial SOS DNA repair system, which comprises a set of DNA repair and cellular survival genes that are induced in response to DNA damage. Its varied DNA binding motifs have been characterized and reported in the Escherichia coli, Bacillus subtilis, rhizobia family members, marine magnetotactic bacterium, Salmonella typhimurium and recently in Mycobacterium tuberculosis and this motifs information has been used in our theoretical analysis to detect its novel regulated genes in radio-resistant Deinococcus radiodurans genome. This bacterium showed presence of SOS-box like consensus sequence in the upstream sequences of 3166 genes with >60% motif score similarity percentage (MSSP) on both strands. Attempts to identify LexA-binding sites and the composition of the putative SOS regulon in D. radiodurans have been unsuccessful so far. To resolve the problem we performed theoretical analysis with modifications on reported data set of genes related to DNA repair (61 genes), stress response (145 genes) and some unusual predicted operons (21 clusters). Expression of some of the predicted SOS-box regulated operon members then was examined through the previously reported microarray data which confirm the expression of only single predicted operon i.e. DRB0143 (AAA superfamily NTPase related to 5-methylcytosine specific restriction enzyme subunit McrB) and DRB0144 (homolog of the McrC subunit of the McrBC restriction modification system). The methodology involved weight matrix construction through CONSENSUS algorithm using information of conserved upstream sequences of eight known genes including dinB, tagC, lexA, recA, uvrB, yneA of B. subtilis while lexA and recA of D. radiodurans through phylogenetic footprinting method and later detection of similar conserved SOS-box like LexA binding motifs through both RSAT & PoSSuMsearch programs. The resultant DNA consensus sequence had highly conserved 14 bp SOS-box like binding site.

摘要

LexA蛋白是细菌SOS DNA修复系统的转录阻遏物,该系统由一组DNA修复和细胞存活基因组成,这些基因在DNA损伤时被诱导表达。其多样的DNA结合基序已在大肠杆菌、枯草芽孢杆菌、根瘤菌家族成员、海洋趋磁细菌、鼠伤寒沙门氏菌中得到表征和报道,最近在结核分枝杆菌中也有相关报道,并且这些基序信息已被用于我们的理论分析,以检测耐辐射球菌基因组中其新的调控基因。该细菌在3166个基因的上游序列中显示出SOS框样共有序列,两条链上的基序得分相似百分比(MSSP)均>60%。到目前为止,在耐辐射球菌中鉴定LexA结合位点和推定的SOS调节子组成的尝试均未成功。为了解决这个问题,我们对与DNA修复(61个基因)、应激反应(145个基因)和一些异常预测操纵子(21个簇)相关的已报道基因数据集进行了修改后的理论分析。然后通过先前报道的微阵列数据检查了一些预测的SOS框调节操纵子成员的表达,这些数据仅证实了单个预测操纵子即DRB0143(与5-甲基胞嘧啶特异性限制酶亚基McrB相关的AAA超家族NTP酶)和DRB0144(McrBC限制修饰系统的McrC亚基的同源物)的表达。该方法包括通过CONSENSUS算法构建权重矩阵,使用包括枯草芽孢杆菌的dinB、tagC、lexA、recA、uvrB、yneA以及耐辐射球菌的lexA和recA在内的八个已知基因的保守上游序列信息,通过系统发育足迹法,随后通过RSAT和PoSSuMsearch程序检测类似的保守SOS框样LexA结合基序。所得的DNA共有序列具有高度保守的14bp SOS框样结合位点。

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