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Regulation of the tryptophan genes in Bacillus

芽孢杆菌色氨酸基因的调控

基本信息

批准号：
7902750
负责人：
PAUL D GOLLNICK
金额：
$ 3.06万
依托单位：
STATE UNIVERSITY OF NEW YORK AT BUFFALO
依托单位国家：
美国
项目类别：
财政年份：
2009
资助国家：
美国
起止时间：
2009-08-14 至 2009-11-30
项目状态：
已结题

项目摘要

DESCRIPTION (provided by applicant): Controlling transcription termination upstream of a coding region is a common strategy to regulate gene expression in bacteria, including many with importance to human health. Such control mechanisms are collectively termed attenuation and antitermination. The proposed research will investigate the mechanisms by which RNA binding proteins recognize and bind to specific sites in RNA, and how these interactions regulate transcription. The mechanism by which protein-protein interactions can modulate the activity of an RNA-binding gene regulatory protein will also be studied. The model system of study is the TRAP protein, an RNA binding protein that regulates transcription attenuation of the tryptophan (trp) genes in Bacillus subtilis and related bacteria. In the presence of excess tryptophan, TRAP is activated to bind to the 5' leader region of the trp operon mRNA and induce formation of a transcription terminator, which halts expression of the genes. TRAP is a unique among characterized RNA binding proteins in that it consists of 11 identical subunits arranged in a ring structure, and in that it binds RNAs that contain up to 11 small (trinucleotide) repeated elements. Recent studies indicate that TRAP binds to RNA by a two-step mechanism. The hypothesis to be tested is that TRAP first binds to the 5'-end of the RNA and then scans until it encounters the 5-most repeats of the binding site, at which point an initiation complex is formed. This is followed by wrapping the remainder of the repeats around the outer perimeter of the protein ring. The detailed mechanism by which TRAP associates with its RNA target will be characterized using a combination of kinetic binding studies, as well as rapid-quench nuclease protection and fluorescence studies. A protein called anti-TRAP (AT) specifically binds to tryptophan-activated TRAP and inhibits it from binding to RNA. Studies will be performed to characterize the structure and function of AT, particularly the mechanism by which it recognizes and binds to TRAP.

描述（由申请人提供）：控制编码区上游的转录终止是调节细菌（包括许多对人类健康具有重要意义的细菌）中基因表达的常见策略。这种控制机制统称为衰减和抗终止。这项研究将研究RNA结合蛋白识别和结合RNA中特定位点的机制，以及这些相互作用如何调节转录。还将研究蛋白质-蛋白质相互作用调节RNA结合基因调节蛋白活性的机制。研究的模型系统是TRAP蛋白，一种调节枯草芽孢杆菌和相关细菌中色氨酸（trp）基因转录衰减的RNA结合蛋白。在过量色氨酸的存在下，TRAP被激活以结合trp操纵子mRNA的5'前导区并诱导转录终止子的形成，其停止基因的表达。TRAP是一种独特的RNA结合蛋白，它由11个相同的亚基组成，排列成环状结构，并且它结合含有多达11个小（三核苷酸）重复元件的RNA。最近的研究表明，TRAP通过两步机制与RNA结合。待测试的假设是TRAP首先结合到RNA的5 '末端，然后扫描，直到它遇到结合位点的最多5个重复，此时形成起始复合物。然后将剩余的重复序列缠绕在蛋白质环的外周上。TRAP与其RNA靶标结合的详细机制将使用动力学结合研究以及快速淬灭核酸酶保护和荧光研究的组合来表征。一种被称为抗TRAP（AT）的蛋白质特异性地结合到聚糖激活的TRAP上，并抑制它与RNA结合。将进行研究以表征AT的结构和功能，特别是其识别和结合TRAP的机制。

项目成果

期刊论文数量（21）

专著数量（0）

科研奖励数量（0）

会议论文数量（0）

专利数量（0）

Substitutions of Thr30 provide mechanistic insight into tryptophan-mediated activation of TRAP binding to RNA.

THR30的取代提供了对色氨酸介导的陷阱结合与RNA的激活的机械洞察力。

DOI：
10.1093/nar/gkl383
发表时间：
2006
期刊：
NUCLEIC ACIDS RESEARCH
影响因子：
14.9
作者：
Payal, Vandana;Gollnick, Paul
通讯作者：
Gollnick, Paul

Bacillus licheniformis Anti-TRAP can assemble into two types of dodecameric particles with the same symmetry but inverted orientation of trimers.

DOI：
10.1016/j.jsb.2010.01.013
发表时间：
2010-04
期刊：
JOURNAL OF STRUCTURAL BIOLOGY
影响因子：
3
作者：
Shevtsov, Mikhail B.;Chen, Yanling;Isupov, Michail N.;Leech, Andrew;Gollnick, Paul;Antson, Alfred A.
通讯作者：
Antson, Alfred A.

Regulation of the tryptophan biosynthetic genes in Bacillus halodurans: common elements but different strategies than those used by Bacillus subtilis.

耐盐芽孢杆菌色氨酸生物合成基因的调控：常见元件，但与枯草芽孢杆菌使用的策略不同。