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来自三种古细菌物种的类recA基因,其推定的蛋白质产物与酿酒酵母的Rad51和Dmc1蛋白相似。

recA-like genes from three archaean species with putative protein products similar to Rad51 and Dmc1 proteins of the yeast Saccharomyces cerevisiae.

作者信息

Sandler S J, Satin L H, Samra H S, Clark A J

机构信息

Department of Molecular and Cell Biology, Division of Genetics, University of California at Berkeley, Berkeley, CA 94720-3202, USA.

出版信息

Nucleic Acids Res. 1996 Jun 1;24(11):2125-32. doi: 10.1093/nar/24.11.2125.

Abstract

The process of homologous recombination has been documented in bacterial and eucaryotic organisms. The Escherichia coli RecA and Saccharomyces cerevisiae Rad51 proteins are the archetypal members of two related families of proteins that play a central role in this process. Using the PCR process primed by degenerate oligonucleotides designed to encode regions of the proteins showing the greatest degree of identity, we examined DNA from three organisms of a third phylogenetically divergent group, Archaea, for sequences encoding proteins similar to RecA and Rad51. The archaeans examined were a hyperthermophilic acidophile, Sulfolobus sofataricus (Sso); a halophile, Haloferax volcanii (Hvo); and a hyperthermophilic piezophilic methanogen, Methanococcus jannaschii (Mja). The PCR generated DNA was used to clone a larger genomic DNA fragment containing an open reading frame (orf), that we refer to as the radA gene, for each of the three archaeans. As shown by amino acid sequence alignments, percent amino acid identities and phylogenetic analysis, the putative proteins encoded by all three are related to each other and to both the RecA and Rad51 families of proteins. The putative RadA proteins are more similar to the Rad51 family (approximately 40% identity at the amino acid level) than to the RecA family (approximately 20%). Conserved sequence motifs, putative tertiary structures and phylogenetic analysis implied by the alignment are discussed. The 5' ends of mRNA transcripts to the Sso radA were mapped. The levels of radA mRNA do not increase after treatment with UV irradiation as do recA and RAD51 transcripts in E.coli and S.cerevisiae. Hence it is likely that radA in this organism is a constitutively expressed gene and we discuss possible implications of the lack of UV-inducibility.

摘要

同源重组过程已在细菌和真核生物中得到记载。大肠杆菌RecA蛋白和酿酒酵母Rad51蛋白是在这一过程中起核心作用的两个相关蛋白家族的典型成员。我们利用由简并寡核苷酸引发的聚合酶链式反应(PCR),这些简并寡核苷酸被设计用于编码蛋白质中显示出最大程度同一性的区域,我们检测了来自第三个系统发育上不同的生物类群——古细菌的三种生物的DNA,以寻找编码与RecA和Rad51相似蛋白质的序列。所检测的古细菌包括嗜热嗜酸菌嗜热栖热菌(Sso)、嗜盐菌沃氏嗜盐栖热菌(Hvo)以及嗜热嗜压产甲烷菌詹氏产甲烷球菌(Mja)。通过PCR产生的DNA被用于克隆包含一个开放阅读框(orf)的更大基因组DNA片段,我们将其称为radA基因,针对这三种古细菌中的每一种都进行了克隆。氨基酸序列比对、氨基酸同一性百分比和系统发育分析表明,这三种古细菌所编码的假定蛋白质彼此相关,并且与RecA和Rad51蛋白家族都相关。假定的RadA蛋白与Rad51家族更为相似(在氨基酸水平上约有40%的同一性),而与RecA家族的相似性较低(约20%)。文中讨论了比对所暗示的保守序列基序、假定的三级结构和系统发育分析。绘制了嗜热栖热菌radA的mRNA转录本的5'端图谱。嗜热栖热菌radA的mRNA水平在紫外线照射处理后不像大肠杆菌和酿酒酵母中的recA和RAD51转录本那样增加。因此,该生物体中的radA可能是一个组成型表达的基因,我们讨论了缺乏紫外线诱导性的可能影响。

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