Developmental Asymmetry in Agrobacterium tumefaciens

根癌农杆菌的发育不对称性

• 批准号: 8572977
• 负责人: Jason Eugene Heindl
• 金额: $ 5.39万
• 依托单位: TRUSTEES OF INDIANA UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2012
• 资助国家: 美国
• 起止时间: 2012-09-01 至 2014-08-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8572977
• 关键词: Address; Animal Model; Animals; Bacteria; Behavior; Binding; Biochemical; Biological Assay; Brucella abortus; Caulobacter crescentus; Cell Cycle Progression; Cells; Coupled; DNA Binding; DNA biosynthesis; Development; Disease; Environment; Eukaryota; Exhibits; Extracellular Structure; Flagella; Genes; Genetic; Genetic Epistasis; Genome; Growth; Growth and Development function; Histidine; Homologous Gene; Hybrids; Individual; Life; Life Style; Mediating; Melilotus; Microbial Biofilms; Microscopy; Modeling; Molecular; Morphogenesis; Morphology; Mutagenesis; Organism; Pathway interactions; Pattern; Phosphorylation; Phosphotransferases; Physiological; Pilum; Plants; Play; Polysaccharides; Process; Prokaryotic Cells; Proteins; Proteobacteria; Regulation; Regulator Genes; Resolution; Rhizobium radiobacter; Role; Series; Shapes; Signal Pathway; Signal Transduction; Sinorhizobium meliloti; Site-Directed Mutagenesis; Stimulus; Surface; System; Techniques; Testing; Type IV Secretion System Pathway; Virulence; appendage; cdc Genes; cell motility; daughter cell; deletion analysis; genetic regulatory protein; innovation; insight; member; novel; pathogen; pathogenic bacteria; protein-histidine kinase; response; retinal rods; sensor; sensor histidine kinase; transcription factor

DESCRIPTION (provided by applicant): Many bacteria, including Agrobacterium tumefaciens, rely on an asymmetric localization, distribution, and orientation of specific appendages when interacting with potential hosts and with their environment. The study of developmental asymmetry is well advanced in the model a-Proteobacterial organism Caulobacter crescentus where the interlinked PleC/DivJ-DivK and CckA-ChpT-CtrA/CpdR His-Asp phosphorelays play a central role. Despite a high degree of conservation among the a-Proteobacteria, including A. tumefaciens little is known about the precise role of these regulatory proteins in these other organisms. Two related proteins, the PleC-DivJ homologue sensor kinases PdhS1 and PdhS2, are restricted to a subset of a-Proteobacteria that frequently associate with a eukaryotic host in a symbiotic or pathogenic relationship. This study examines the role of these regulatory proteins in A. tumefaciens. In particular, this application addresses how PdhS1 and PdhS2 contribute to the morphological, physiological, and temporal development of A. tumefaciens and whether these processes contribute to the transition between host and non-host niches. This will be performed using a combination of genetic, biochemical, and cytological analyses. PdhS1 and PdhS2 will both be biochemically dissected to determine the contribution of individual catalytic and regulatory domains to their activity and to identify potential interacting partners. A. tumefaciens binds to abiotic and biotic surfaces in a polar orientation and is known to elaborate polar flagella, pili, and a type IV secretion system. In addition, A. tumefaciens and many other a-Proteobacteria divide by an asymmetric process of budding, proceeding through an attached, non-motile form to a morphologically distinct motile form. Phenotypic analysis will evaluate the contribution of PdhS1 and PdhS2 to polar development, motility, biofilm formation, and virulence. Additional genes and regulatory interactions that influence coordination of polar morphogenesis, motility, and biofilm formation in A. tumefaciens will be identified using a combination of transposon mutagenesis and a novel high-throughput microscopy screen.

描述（由申请人提供）：许多细菌，包括农杆菌，在与潜在宿主及其环境相互作用时，都依赖于特定附属物的不对称定位，分布和方向。在模型A蛋白细菌生物的caulobacter crescentus中，发育不对称的研究非常先进，其中相互链接的PLEC/DIVJ-DIVK和CCKA-CCA-CHPT-CTRA/CPDR HIS-ASP HIS-ASP Phosphorelays起着核心作用。尽管a蛋白蛋白蛋白菌具有高度的保护，但包括曲霉曲霉的人数对这些调节蛋白在这些其他生物中的确切作用知之甚少。两种相关的蛋白质，PLEC-DIVJ同源传感器激酶PDHS1和PDHS2，仅限于a-蛋白细菌的一个子集，该子蛋白经常与共生或致病关系中的真核宿主相关。这项研究检查了这些调节蛋白在曲霉杆菌中的作用。特别是，该应用程序解决了PDHS1和PDHS2如何促进曲霉曲霉的形态，生理和时间发育，以及这些过程是否有助于宿主和非宿主壁球之间的过渡。这将使用遗传，生化和细胞学分析的组合进行。 PDHS1和PDHS2都将在生化上剖析，以确定单个催化和调节域对其活性的贡献并确定潜在的相互作用伙伴。 A. tumefaciens在极性方向上与非生物和生物表面结合，并且已知可以精细的极性鞭毛，pili和IV型分泌系统。此外，曲霉曲霉和许多其他A-蛋白细菌除以萌芽过程，通过附着的非运动形式来进行形态上不同的运动形式。表型分析将评估PDHS1和PDHS2对极性发育，运动，生物膜形成和毒力的贡献。将使用转座子诱变和新型的高通量显微镜筛选的组合来鉴定影响曲霉曲霉中极性形态发生，运动性和生物膜形成协调的其他基因和调节相互作用。