Myosins VIIA, VIIB & XV in Fly Hearing and Morphogenesis

肌球蛋白VIIA、VIIB

• 批准号: 7252027
• 负责人: DANIEL PETER KIEHART
• 金额: $ 33.03万
• 依托单位: DUKE UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2005
• 资助国家: 美国
• 起止时间: 2005-07-15 至 2010-06-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7252027
• 关键词: Actins; Address; Adhesions; Adult; Affinity; Animals; Auditory; Binding; Biological Models; Biological Process; Biology; Cell physiology; Cells; Class; Complex; Cytoskeleton; Development; Drosophila genus; Drosophila melanogaster; Electron Microscopy; Elements; Embryo; Gene Proteins; Genetic; Hair; Hearing; Heart; Human; In Vitro; Individual; Interdisciplinary Study; Loa; Mediating; Membrane; Methods; Microfilaments; Microtubules; Molecular; Molecular Genetics; Molecular Machines; Morphogenesis; Motor; Mus; Mutate; Mutation; Myosin ATPase; Numbers; Organ; Orthologous Gene; Phenotype; Physiology; Play; Process; Proteins; Recombinant Proteins; Research Personnel; Resolution; Role; SH3 Domains; Sensory; Signal Transduction; Specific qualifier value; Structure; System; Tail; Work; Yeasts; base; design; fly; hybrid protein; in vivo; insight; interdisciplinary approach; member; novel; programs; protein function; protein structure; protein structure function; relating to nervous system; research study; sound; success; tool; transcription factor; yeast two hybrid system

DESCRIPTION (provided by applicant): We propose a multidisciplinary research program that investigates the function of protein complexes at the heart of auditory reception in the model system, Drosophila melanogaster. The biology of hearing is astonishingly conserved between humans and fly: both the transcription factors that specify the development of organs of auditory reception and the molecular machines that transduce sound into neural signals are highly conserved. In both systems, cellular differentiation for the perception of sound requires elaborate subcellular arrangements of cytoskeletal elements, including actin filaments and microtubules. Mutations in the human and mouse orthologs of ck/myoVIIA and 10A/myoXV demonstrate the key roles these proteins play in auditory reception and vestibular function, but the molecular mechanism(s) by which these motor proteins contribute to these processes remain controversial. We propose an interdisciplinary approach to structure/function analysis of ck/myoVIIA, 28B/myoVIIB and 10A/myoXV (together, the myosin VII subfamily) in Drosophila where we use uniquely powerful molecular genetic tools to relate protein structure-function studies in vitro to structure/function analyses in vivo. We have already shown that ck/myoVIIA mutates to lethality, is required to drive changes in the structure of the actin cytoskeleton during morphogenesis and is required for auditory reception in adult flies. We have demonstrated that 10A/myoXV is essential for fly viability. We propose three specific aims 1) that address the cellular mechanisms of motor protein function; 2) that identify the binding partners with which these motor proteins interact; and 3) elucidate the structure of their cargo binding tails. Our specific aims will inform one another, but do not depend on one another's success. Together, our studies offer unique opportunities to probe the supramolecular protein complexes that require motor protein function and that drive cellular function in auditory reception, vestibular function and morphogenesis.

描述（由申请人提供）：我们提出了一项多学科研究计划，研究黑腹果蝇模型系统中听觉接收核心的蛋白质复合物的功能。人类和苍蝇之间的听觉生物学惊人地保守：指定听觉接收器官发育的转录因子和将声音转换为神经信号的分子机器都高度保守。在这两个系统中，感知声音的细胞分化需要细胞骨架元件的精细亚细胞排列，包括肌动蛋白丝和微管。 ck/myoVIIA 和 10A/myoXV 的人和小鼠直系同源物的突变证明了这些蛋白质在听觉接收和前庭功能中发挥的关键作用，但这些运动蛋白促进这些过程的分子机制仍然存在争议。我们提出了一种跨学科方法对果蝇中的 ck/myoVIIA、28B/myoVIIB 和 10A/myoXV（统称为肌球蛋白 VII 亚家族）进行结构/功能分析，其中我们使用独特强大的分子遗传学工具将体外蛋白质结构功能研究与体内结构/功能分析。我们已经表明，ck/myoVIIA 突变为致死性，是在形态发生过程中驱动肌动蛋白细胞骨架结构变化所必需的，也是成年果蝇听觉接收所必需的。我们已经证明 10A/myoXV 对于果蝇生存能力至关重要。我们提出了三个具体目标：1）解决运动蛋白功能的细胞机制； 2) 识别与这些运动蛋白相互作用的结合伙伴； 3）阐明其货物绑定尾部的结构。我们的具体目标将相互告知，但并不取决于彼此的成功。总之，我们的研究提供了独特的机会来探测需要运动蛋白功能并驱动听觉接收、前庭功能和形态发生中的细胞功能的超分子蛋白复合物。