Mechanisms of Anthrax Virulence Factor AtxA.

炭疽毒力因子 AtxA 的机制。

• 批准号: 8204682
• 负责人: EVGENY A NUDLER
• 金额: $ 21.13万
• 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 2013-02-28
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8204682
• 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; public health relevance; 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。尽管其毒力作用已在基因上得到证实，但其作用机制细节仍不清楚。我们发现其产物AtxA在体内与炭疽RNA聚合酶结合，并在体外证明了其转录活性。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且在许多方面独特的经验，我们提出了 AtxA 结构和功能研究的全面计划。作为拟议研究的结果，将详细了解 AtxA 在炭疽毒素表达中的作用，并深入研究其作用的分子机制，彻底分析该因子的差异磷酸化与炭疽毒素之间的相互作用。它的转录活性，以及​​该因子在“ON”和“OFF”状态下的高分辨率结构。总而言之，这些见解将有助于更好地了解炭疽毒力和致病性，同时作为基于 AtxA 的炭疽毒素抑制剂的结构设计的起点，该抑制剂可抑制这种重要的人畜共患病原体和 A 类制剂产生炭疽毒素。 公共卫生相关性：炭疽杆菌是人畜共患炭疽感染的常见原因，并已在美国本土被用作生物恐怖武器。我们提出了对炭疽毒力和毒素产生的主要调控因子转录因子AtxA进行生化和结构研究的全面计划。作为这项研究的结果，将出现对 AtxA 作用机制的详细了解，同时将收集必要的结构和生化信息以对其进行评估，作为基于知识的药物设计的目标。