Regulation of the tryptophan genes in Bacillus

芽孢杆菌色氨酸基因的调控

• 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
• 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
• 来源: https://reporter.nih.gov/project-details/7902750
• 关键词: Affect; Bacillus (bacterium); Bacillus subtilis; Bacteria; Binding; Binding Sites; Biochemical; Biological Models; Code; Collaborations; Complex; Elements; Fluorescence; Gene Expression; Genes; Genetic; Genetic Transcription; Goals; Health; Human; Kinetics; Messenger RNA; Operon; Proteins; RNA; RNA Binding; RNA-Binding Proteins; Regulation; Research; Resolution; Scanning; Site; Structure; Surface Plasmon Resonance; System; Testing; Thermodynamics; Transfer RNA; Transfer RNA Aminoacylation; Translations; Trinucleotide Repeats; Tryptophan; antitermination; attenuation; genetic regulatory protein; novel; nuclease; protein protein interaction; response; stopped-flow fluorescence; transcription termination

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）：编码区域上游的转录终止是调节细菌基因表达的常见策略，其中包括许多对人类健康重要性的表达。这样的控制机制统称为衰减和抗衰测。拟议的研究将研究RNA结合蛋白识别并结合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相互作用的机制调节RNA结合基因调节蛋白的活性。研究模型系统是TRAP蛋白，一种RNA结合蛋白，可调节枯草芽孢杆菌和相关细菌中色氨酸（TRP）基因的转录衰减。在存在过多的色氨酸的情况下，陷阱被激活与TRP操纵子mRNA的5'领导区域结合并诱导转录终结子的形成，该转录终结子停止了基因的表达。陷阱是特征性的RNA结合蛋白中的独特之处，因为它由11个相同的亚基组成，该亚基在环结构中排列，并且它结合了最多包含11个小（三核苷酸）重复元件的RNA。最近的研究表明，陷阱通过两步机制与RNA结合。要测试的假设是，陷阱首先结合了RNA的5'末端，然后扫描直到遇到结合位点的5个重复序列，此时形成了启动复合物。接下来是将剩余的重复序列包裹在蛋白环的外周围。将诱捕器与其RNA靶标相关联的详细机制将通过动力学结合研究以及快速猝灭的核酸酶保护和荧光研究来表征。一种称为抗陷阱（AT）的蛋白质特异性结合了色氨酸激活的诱捕器，并抑制其与RNA结合。将进行研究以表征AT的结构和功能，尤其是识别并结合陷阱的机制。

项目成果

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

• 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。

• DOI: 10.1128/jb.186.3.818-828.2004
• 发表时间: 2004
• 期刊: Journal of bacteriology
• 影响因子: 3.2
• 作者: Szigeti,Reka;Milescu,Mirela;Gollnick,Paul
• 通讯作者: Gollnick,Paul

Cellular levels of trp RNA-binding attenuation protein in Bacillus subtilis.

枯草芽孢杆菌中 trp RNA 结合衰减蛋白的细胞水平。

• DOI: 10.1128/jb.186.15.5157-5159.2004
• 发表时间: 2004
• 期刊: Journal of bacteriology.
• 作者: McCabe,BarbaraC;Gollnick,Paul
• 通讯作者: Gollnick,Paul

Mechanism for pH-dependent gene regulation by amino-terminus-mediated homooligomerization of Bacillus subtilis anti-trp RNA-binding attenuation protein.

通过枯草芽孢杆菌抗色氨酸 RNA 结合衰减蛋白的氨基末端介导的同源寡聚化实现 pH 依赖性基因调节的机制。

• DOI: 10.1073/pnas.1004981107
• 发表时间: 2010
• 期刊: Proceedings of the National Academy of Sciences of the United States of America
• 影响因子: 11.1
• 作者: Sachleben,JosephR;McElroy,CraigA;Gollnick,Paul;Foster,MarkP
• 通讯作者: Foster,MarkP