Class I myosins regulate protrusive forces at the actin-membrane interface

I 类肌球蛋白调节肌动蛋白-膜界面的突出力

• 批准号: 10612046
• 负责人: Mira Krendel
• 金额: $ 53.4万
• 依托单位: UPSTATE MEDICAL UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2021
• 资助国家: 美国
• 起止时间: 2021-07-06 至 2025-04-30
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10612046
• 关键词: Actins; Adhesions; Affect; Automobile Driving; Binding; Biological Models; Biological Process; Cell Shape; Cell membrane; Cell model; Cells; Complex; Cytoskeleton; Data; Endocytic Vesicle; Endocytosis; Environment; Eukaryota; Experimental Models; Filament; Fission Yeast; Generations; Immune response; Invaded; Knockout Mice; Macrophage; Mammalian Cell; Measures; Mediating; Membrane; Microfilaments; Modeling; Molecular; Molecular Motors; Motor; Motor Activity; Mus; Mutation; Myosin ATPase; Myosin Type I; Nonmuscle Myosin Type IIA; Nutrient; Phagocytes; Phagocytosis; Physiological; Play; Plus End of the Actin Filament; Polymers; Proteins; Regulation; Regulation of Cell Shape; Resistance; Role; Shapes; Site; Structure; System; Tail; Techniques; Testing; Work; Yeast Model System; Yeasts; cell growth; cell motility; experimental study; extracellular; migration; monomer; mutant; pathogen; polymerization; pressure; protein crosslink; recruit

In this project, we will dissect the roles of class I myosins, molecular motors that bind actin filaments through their motor domains and plasma membrane through their tail domains, in actin-mediated cell shape changes. We will test the hypothesis that myosin I motor activity promotes assembly of branched actin networks at the sites of membrane deformation, including endocytic vesicles, phagocytic cups, and podosomes. The proposed work will use two model systems: fission yeast, where myosin I is required for endocytosis, and murine macrophages, which rely on myosin I for phagocytosis and cell migration. In aim 1, we will determine how myosin motor activity contributes to endocytic actin patch assembly and internalization in yeast. We will use the genetically tractable yeast system to express mutant myosin from the endogenous myosin locus and apply molecule counting techniques, which are well established in the yeast endocytosis system, to measure the rates of actin assembly. In aim 2, we will test the role of myosin motor activity in actin and membrane dynamics during phagocytosis and cell migration. These studies will take advantage of the genetically modified mice lacking class I myosins that are available in our labs. In aim 3, we will determine how myosin-membrane interactions are regulated to contribute to myosin functions in actin assembly and immune response. The proposed work will shed light on the molecular mechanisms driving actin- dependent membrane deformation.

在这个项目中，我们将剖析I类肌球蛋白的作用，即结合肌动蛋白的分子电机 通过其运动域和质膜的细丝通过其尾部域， 肌动蛋白介导的细胞形状变化。我们将测试肌球蛋白I运动活动的假设 促进在膜变形部位的分支肌动蛋白网络组装，包括 内吞囊泡，吞噬杯和足体。拟议的工作将使用两种模型 系统：裂变酵母，肌球蛋白I所需要的内吞作用和鼠巨噬细胞需要 依赖肌球蛋白I用于吞噬作用和细胞迁移。在AIM 1中，我们将确定如何 肌球蛋白运动活性有助于酵母中内吞肌动蛋白斑块的组装和内在化。 我们将使用遗传处理的酵母系统来表达从 内源性肌球蛋白基因座并应用分子计数技术，这些技术已建立得很好 在酵母内吞作用系统中，以测量肌动蛋白组装的速率。在AIM 2中，我们将测试 肌球蛋白运动活性在吞噬作用和细胞中肌动蛋白和膜动力学中的作用 迁移。这些研究将利用缺乏I类的转基因小鼠 我们实验室中可用的肌球蛋白。在AIM 3中，我们将确定肌球蛋白膜如何 相互作用受到调节，以促进肌动蛋白组装中的肌球蛋白功能和免疫 回复。拟议的工作将阐明驱动肌动蛋白的分子机制 依赖性膜变形。

项目成果

Building the phagocytic cup on an actin scaffold.

• DOI: 10.1016/j.ceb.2022.102112
• 发表时间: 2022-08
• 期刊: CURRENT OPINION IN CELL BIOLOGY
• 影响因子: 7.5
• 作者: Krendel, Mira;Gauthier, Nils C.
• 通讯作者: Gauthier, Nils C.

Cappin' or formin': Formin and capping protein competition for filament ends shapes actin networks.

• DOI: 10.1083/jcb.202302009
• 发表时间: 2023-04-03
• 期刊: The Journal of cell biology

Phagocytic 'teeth' and myosin-II 'jaw' power target constriction during phagocytosis.

• DOI: 10.7554/elife.68627
• 发表时间: 2021-10-28
• 期刊: eLife
• 影响因子: 7.7
• 作者: Vorselen D;Barger SR;Wang Y;Cai W;Theriot JA;Gauthier NC;Krendel M
• 通讯作者: Krendel M

A theoretical model of efficient phagocytosis driven by curved membrane proteins and active cytoskeleton forces.

由弯曲膜蛋白和活跃细胞骨架力驱动的高效吞噬作用的理论模型。

• DOI: 10.1039/d2sm01152b
• 发表时间: 2022
• 期刊: Soft matter
• 影响因子: 3.4
• 作者: Sadhu,RajKumar;Barger,SarahR;Penič,Samo;Iglič,Aleš;Krendel,Mira;Gauthier,NilsC;Gov,NirS
• 通讯作者: Gov,NirS

F-actin organization and target constriction during primary macrophage phagocytosis is balanced by competing activity of myosin-I and myosin-II.

• DOI: 10.1091/mbc.e22-06-0210
• 发表时间: 2022-12-01
• 期刊: MOLECULAR BIOLOGY OF THE CELL
• 影响因子: 3.3
• 作者: Barger, Sarah R.;Vorselen, Daan;Gauthier, Nils C.;Theriot, Julie A.;Krendel, Mira
• 通讯作者: Krendel, Mira