Role of myosin-1 in membrane and actin dynamics

myosin-1 在膜和肌动蛋白动力学中的作用

• 批准号: 6835864
• 负责人: CHRISTINA A KING-SMITH
• 金额: $ 4.45万
• 依托单位: UNIVERSITY OF PENNSYLVANIA
• 依托单位国家: 美国
• 财政年份: 2004
• 资助国家: 美国
• 起止时间: 2004-06-16 至 2006-06-15
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/6835864
• 关键词: actins; cell line; cell membrane; cell motility; chemical kinetics; immunocytochemistry; membrane activity; molecular biology; myosins; neoplastic cell culture for noncancer research; protein isoforms; protein localization; protein protein interaction; protein structure function; small interfering RNA; actins; cell line; cell membrane; cell motility; chemical kinetics; immunocytochemistry; membrane activity; molecular biology; myosins; neoplastic cell culture for noncancer research; protein isoforms; protein localization; protein protein interaction; protein structure function; small interfering RNA

DESCRIPTION (provided by applicant): The objectives of the proposed research are to understand the roles of myosin I in membrane retraction and retrograde actin flow. Myosin Is represent one of the largest classes of unconventional myosins in humans, yet little is known about their activity, distribution, or regulation in vertebrate cells. Specific aims include the following: 1) To characterize the distribution of myo1c during lamellipodial extension and retraction in MTLn3 cells, a metastatic mammalian cell line. This will be accomplished by immunofluorescence microscopy and transient expression of eGFP-myosin fusion proteins. 2) To determine the effect of myosin-I "knockdown" on cell spreading, membrane retraction, and retrograde actin flow using small interfering RNA (siRNA). Retrograde flow in control and myosin-I knockdown cells will be assessed by motility of surface-attached latex beads. 3) To determine the relationship between the mechanical activity of myo1c and membrane dynamics. We will express myo1c mutants that have altered motor properties, and we will examine the effect of these mutants on membrane dynamics.

描述（由申请人提供）： 拟议的研究的目标是了解肌球蛋白I在膜回缩和逆行肌动蛋白流动中的作用。肌球蛋白是人类中最大的非常规肌球蛋白的类别之一，但对它们在脊椎动物细胞中的活性，分布或调节知之甚少。具体目的包括以下目的：1）表征在MTLN3细胞（转移性哺乳动物细胞系MTLN3细胞）中膜状延伸过程中MyO1c的分布。这将通过免疫荧光显微镜和EGFP肌球蛋白融合蛋白的瞬时表达来实现。 2）使用小干扰RNA（siRNA）确定肌球蛋白1“敲低”对细胞扩散，膜回缩和逆行肌动蛋白流的影响。 对照和肌球蛋白I敲除细胞的逆行流将通过表面连接的乳胶珠的运动来评估。 3）确定MyO1c和膜动力学的机械活性之间的关系。我们将表达改变运动特性的MyO1c突变体，并将检查这些突变体对膜动力学的影响。