Mechanisms of Anthrax Virulence Factor AtxA.

炭疽毒力因子 AtxA 的机制。

• 批准号: 8029037
• 负责人: EVGENY A NUDLER
• 金额: $ 25.35万
• 依托单位: NEW YORK UNIVERSITY SCHOOL OF MEDICINE
• 依托单位国家: 美国
• 财政年份: 2010
• 资助国家: 美国
• 起止时间: 2010-12-15 至 2012-11-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8029037
• 关键词: Address; Anthrax disease; Bacillus anthracis; Binding; Biochemical; Categories; Complex; DNA-Directed RNA Polymerase; Data; Drug Delivery Systems; Drug Design; Enzymes; Escherichia coli; Evaluation; Gene Expression; Genes; Genetic Transcription; In Vitro; Infection; Investigation; Mass Spectrum Analysis; Methods; Molecular; Molecular Conformation; Pathogenesis; Pathogenicity; Phosphorylation; Plasmids; Play; Production; Protein Dephosphorylation; Proteins; Regulation; Regulator Genes; Research; Resolution; Role; Soil; Solutions; Staging; Structure; System; Toxin; Variant; Virulence; Virulence Factors; anthrax toxin; base; design; experience; his6 tag; in vitro activity; in vivo; inhibitor/antagonist; insight; knowledge base; light scattering; new technology; pathogen; tool; transcription factor; weapons

DESCRIPTION (provided by applicant): Bacillus anthracis plasmid-borne gene atxA encodes a major regulator of anthrax toxin expression. Although its role in virulence has been confirmed genetically, the mechanistic details of its action remain obscure. We have discovered that its product, AtxA, binds to anthrax RNA polymerase in vivo, and demonstrated its transcriptional activity in vitro. Using our extensive and in many ways unique experience in elucidating structural and biochemical mechanisms of various bacterial regulators of gene expression we propose a comprehensive plan of AtxA structural and functional research. As a result of the proposed research a detailed understanding of AtxA role in the expression of the anthrax toxin will emerge, complete with in-depth investigation of the molecular mechanism of its action, thorough analysis of the interplay between the differential phosphorylation of this factor and its transcriptional activity, and a high resolution structure of this factor in the "ON" and "OFF" states. Altogether these insights would allow for a better understanding of anthrax virulence and pathogenicity, while serving as a starting point of the structure-based design of AtxA-based inhibitors of anthrax toxin production by this important zoonotic pathogen and a Category A agent. PUBLIC HEALTH RELEVANCE: Bacillis anthracis is a routine cause of zoonotic anthrax infections and has been used as a bioterrorist weapon on the US soil. We propose a comprehensive plan of biochemical and structural research of the main regulator of anthrax virulence and toxin production, transcription factor AtxA. As the result of this research, a detailed understanding of AtxA mechanism of action will emerge while necessary structural and biochemical information will be gathered for its evaluation as a target for knowledge-based drug design.

描述（由申请人提供）：炭疽芽孢杆菌质粒携带的基因atxA编码炭疽毒素表达的主要调节因子。尽管其在毒力中的作用已被遗传学证实，但其作用的机制细节仍不清楚。我们发现其产物AtxA在体内与炭疽RNA聚合酶结合，并在体外证明其转录活性。利用我们在阐明各种细菌基因表达调节因子的结构和生化机制方面的广泛和独特的经验，我们提出了一个全面的AtxA结构和功能研究计划。通过本研究，我们将详细了解AtxA在炭疽毒素表达中的作用，深入研究其作用的分子机制，深入分析该因子的差异磷酸化与其转录活性之间的相互作用，并获得该因子在“开”和“关”状态下的高分辨率结构。总的来说，这些见解将有助于更好地了解炭疽的毒力和致病性，同时作为基于结构设计的基于atxa的炭疽毒素抑制剂的起点，这种重要的人畜共患病原体和a类病原体产生炭疽毒素。