Unconventional Myosins of the Hair Cell

毛细胞的非常规肌球蛋白

• 批准号: 6959257
• 负责人: RYAN L KARCHER
• 金额: $ 4.83万
• 依托单位: MC LAUGHLIN RESEARCH INSTITUTE
• 依托单位国家: 美国
• 财政年份: 2004
• 资助国家: 美国
• 起止时间: 2004-09-30 至 2007-09-29
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/6959257
• 关键词: actins; binding sites; cell differentiation; chemical binding; developmental neurobiology; ear hair cell; embryonic stem cell; endocytosis; gene expression; genetic regulation; genetically modified animals; hearing disorders; inhibitor /antagonist; intermolecular interaction; laboratory mouse; molecular genetics; mutant; myosins; neurogenetics; newborn animals; nucleotide analog; postdoctoral investigator; protein localization; protein structure function; site directed mutagenesis

DESCRIPTION (provided by applicant): Unconventional myosins comprise an important class of molecular motor proteins. They are involved in a myriad of different functions in eukaryotes, including transport of vesicles and mRNAs, actin reorganization, and predominantly structural functions. Several myosins (Ic, VI, VIla, and XV) have essential roles in hair-cell function. To ascertain the specific cellular roles of these respective motors in hair cells, a chemical-genetic approach (already validated for myosin-1c in slow adaptation) will be utilized. Mutant myosins will be engineered to enlarge their ATP binding pockets, which will sensitize them to synthetic ADP analogs. Knock-in or transgenic mice expressing these sensitized motors will allow the study of each motor's hypothesized role(s) in slow adaptation, actin bundle development, or apical endocytosis. This approach provides several advantages; foremost of which is tight temporal control over the specific, selective inhibition of myosin function. The ability to selectively 'lock down' a specific myosin onto actin filaments (rigor complex) enables a more rigorous determination of in vivo function without the artifacts caused by overexpression.

描述（由申请人提供）：非常规肌球蛋白包括一类重要的分子马达蛋白。它们参与真核生物中的无数不同功能，包括囊泡和mRNA的运输，肌动蛋白重组和主要的结构功能。几种肌球蛋白（Ic，VI，VIla和XV）在毛细胞功能中起重要作用。为了确定毛细胞中这些相应马达的特定细胞作用，将利用化学遗传方法（已在缓慢适应中验证肌球蛋白-1c）。突变肌球蛋白将被工程化以扩大它们的ATP结合口袋，这将使它们对合成的ADP类似物敏感。表达这些致敏马达的敲入或转基因小鼠将允许研究每个马达在缓慢适应、肌动蛋白束发育或顶端内吞作用中的假设作用。这种方法提供了几个优点，其中最重要的是严格的时间控制特异性，选择性抑制肌球蛋白功能。选择性地将特定肌球蛋白“锁定”到肌动蛋白丝上（僵硬复合物）的能力使得能够更严格地确定体内功能，而没有由过表达引起的伪影。