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FruA,一种对黄色黏球菌发育至关重要的假定转录因子。

FruA, a putative transcription factor essential for the development of Myxococcus xanthus.

作者信息

Ogawa M, Fujitani S, Mao X, Inouye S, Komano T

机构信息

Department of Biology, Tokyo Metropolitan University, Japan.

出版信息

Mol Microbiol. 1996 Nov;22(4):757-67. doi: 10.1046/j.1365-2958.1996.d01-1725.x.

Abstract

A new developmental gene, fruA, of Myxococcus xanthus was cloned using a one-step cloning vector, TnV. DNA sequencing of the wild-type allele of the fruA gene indicated that the fruA gene encodes a protein of 229 amino acid residues with a calculated molecular weight of 24672. The deduced amino acid sequence of FruA protein showed similarity to those of many bacterial regulatory proteins carrying a DNA-binding helix-turn-helix motif. The transcription-initiation site of the fruA gene was determined by a primer-extension experiment. Development of M. xanthus cells with a disrupted fruA gene stopped at the stage of mound formation. Although cells were able to aggregate to form mounds, myxospores were not formed. By Northern and Western blot analysis, it was found that the fruA expression was not detected during vegetative growth but initiated at around 6 h and reached the highest level at 12 h after the onset of development. Expression of the fruA gene was dependent on the expression of asg, bsg, csg, dsg, and esg genes, indicating that a series of intercellular signalling is necessary for the expression of the fruA gene. The effects of the fruA mutation on beta-galactosidase expression of various developmentally regulated genes fused with the lacZ gene were analysed; three developmental lacZ fusions (omega 4469, omega 4273 and omega 4500) were either poorly induced or not induced at all, while three other lacZ fusions (omega 4408, omega 4521 and omega 4455) expressed at the early stage of development were normally induced but were unable to be repressed at a later stage of development as in the wild-type strain. Interestingly, in the fruA mutant, tps (the gene for protein S) was not activated. From these results together with analysis of the amino acid sequence of FruA, we propose that FruA is a putative transcription factor required for the development of M.xanthus.

摘要

利用一步克隆载体TnV克隆了黄色粘球菌的一个新的发育基因fruA。fruA基因野生型等位基因的DNA测序表明,fruA基因编码一个由229个氨基酸残基组成的蛋白质,计算分子量为24672。推导的FruA蛋白氨基酸序列与许多带有DNA结合螺旋-转角-螺旋基序的细菌调节蛋白的序列相似。通过引物延伸实验确定了fruA基因的转录起始位点。fruA基因被破坏的黄色粘球菌细胞的发育在土堆形成阶段停止。虽然细胞能够聚集形成土堆,但不能形成粘孢子。通过Northern和Western印迹分析发现,fruA在营养生长期间未检测到表达,但在发育开始后约6小时开始表达,并在12小时达到最高水平。fruA基因的表达依赖于asg、bsg、csg、dsg和esg基因的表达,表明一系列细胞间信号传导对于fruA基因的表达是必需的。分析了fruA突变对与lacZ基因融合的各种发育调控基因的β-半乳糖苷酶表达的影响;三个发育性lacZ融合体(omega 4469、omega 4273和omega 4500)诱导不佳或根本不被诱导,而另外三个在发育早期表达的lacZ融合体(omega 4408、omega 4521和omega 4455)正常诱导,但在发育后期不能像野生型菌株那样被抑制。有趣的是,在fruA突变体中,tps(蛋白S的基因)未被激活。综合这些结果以及对FruA氨基酸序列的分析,我们提出FruA是黄色粘球菌发育所需的一个假定转录因子。

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