Enzymatic and Motor Properties of Myosin 19

肌球蛋白 19 的酶学和运动特性

基本信息

批准号：
8517048
负责人：
Omar Alberto Quintero-Carmona
金额：
$ 11.14万
依托单位：
UNIVERSITY OF RICHMOND
依托单位国家：
美国
项目类别：
财政年份：
2011
资助国家：
美国
起止时间：
2011-08-02 至 2016-02-29
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/8517048
关键词：
ATP phosphohydrolase Actins Antibodies Apoptosis Apoptotic Applications Grants Biochemical Biological Assay Calcium Cancer cell line Cell Culture Techniques Cell Cycle Cell Cycle Progression Cell Cycle Stage Cell Extracts Cell Line Cell Volumes Cell division Cell physiology Cells Characteristics Cultured Cells DNA Sequence Rearrangement Data Analyses Disease Ectopic Expression Equilibrium Faculty Filament Funding Genes Goals Health Human In Vitro Induction of Apoptosis Insecta Institution Kinetics Lead Length Life Literature Location Malignant Neoplasms Malignant neoplasm of lung Mentored Research Scientist Development Award Mentors Mentorship Metabolism Microscopy Microtubules Mitochondria Mitosis Mitotic Mitotic spindle Modeling Molecular Motors Motor Motor Activity Mutation Myosin ATPase Neck Neurons Normal tissue morphology Organelles Pathway interactions Phenotype Play Process Production Program Development Property Protein Sequence Analysis Proteins RNA Interference Regulation Research Research Project Grants Research Training Role Science Sequence Analysis Slide Students System Tail Time Training Transfection Vertebrates base career cell motility gain of function human disease in vivo knock-down mitochondrial dysfunction novel overexpression professor research study segregation skills undergraduate research

项目摘要

DESCRIPTION (provided by applicant): The long-term goal of this research project is to understand the enzymatic properties, motor properties, and cellular functions of myosin XIX (Myo19), a molecular motor protein involved in the transport and localization of mitochondria, an organelle central to multiple cellular processes. Mitochondrial dynamics have been shown to be partially actin-based, and mutations in other genes involved in mitochondrial dynamics lead to human diseases. Additionally, microarray analyses indicate that Myo19 expression is increased in certain cancers, compared to normal tissues. Previous studies show that Myo19 plays a role in mitochondrial motility in cultured cells as expression of GFP-tagged Myo19 constructs alters mitochondrial dynamics in a pulmonary cancer cell line and in a cultured neuronal cell line. Taken together, analysis of the Myo19 protein sequence and analysis of the effect of GFP-Myo19 expression in cultured cells lead to the hypothesis that Myo19 plays a role in mitochondrial dynamics, as it contains a conserved myosin motor domain and interacts with mitochondria via its tail domain. We will use both Myo19 knock-down and overexpression of functional and non-functional Myo19 constructs in cell culture models of cell division and apoptosis to determine what role Myo19 may play in each of those processes. Expression and purification of Myo19 motor domain will allow for a complete in vitro analysis of the rate and equilibrium constants associated with the motor properties of Myo19. Additionally, purified protein will be used to directly examine the motor properties of Myo19 using an in vitro actin sliding filament motility assay system. Examination of both the enzymatic and motor characteristics of Myo19 in vitro will allow the determination of the mechanism by which Myo19 motor activity generates force, and determination of factors that may regulate its function in vivo. By examining the functional roles, enzymatic properties, and motor properties, of Myo19, these proposed studies will further our understanding of mitochondrial dynamics in health and disease.

描述（由申请人提供）：该研究项目的长期目标是了解肌球蛋白XIX（MyO19）的酶促性能，运动性能和细胞功能，这是一种参与线粒体传输和定位的分子运动蛋白，这是一种中心的多个细胞过程。线粒体动力学已被证明是部分基于肌动蛋白的，并且与线粒体动力学有关的其他基因中的突变导致人类疾病。此外，微阵列分析表明，与正常组织相比，某些癌症的肌酸表达增加。先前的研究表明，MyO19在培养细胞中的线粒体运动中起作用，因为GFP标记的MyO19构建的表达会在肺癌细胞系和培养的神经元细胞系中改变线粒体动力学。综上所述，对MyO19蛋白序列的分析以及GFP-MYO19表达在培养细胞中的影响的分析导致假说，MyO19在线粒体动力学中起着作用，因为它包含一个保守的肌球蛋白运动结构域，并通过其尾部结构域与线粒体相互作用。我们将同时使用MyO19敲低和过表达在细胞分裂和细胞凋亡的细胞培养模型中的功能和非功能性myo19构建体，以确定MyO19在每个过程中可能扮演什么作用。 MyO19运动结构域的表达和纯化将允许对与MyO19的运动性能相关的速率和平衡常数进行完整的体外分析。另外，纯化的蛋白质将用于使用体外肌动蛋白滑动丝运动测定系统直接检查MyO19的运动特性。在体外检查MyO19的酶促和运动特性，将允许确定MyO19运动活性产生力的机制，并确定可能调节其在体内功能的因子。通过检查Myo19的功能作用，酶特性和运动性能，这些提出的研究将进一步了解我们对健康和疾病中线粒体动力学的理解。

项目成果

期刊论文数量（5）

专著数量（0）

科研奖励数量（0）

会议论文数量（0）

专利数量（0）

Permeabilization activated reduction in fluorescence: A novel method to measure kinetics of protein interactions with intracellular structures.

透化激活荧光减少：一种测量蛋白质与细胞内结构相互作用动力学的新方法。

DOI：
10.1002/cm.21306
发表时间：
2016
期刊：
Cytoskeleton (Hoboken, N.J.)
影响因子：
0
作者：
Singh,PaliP;Hawthorne,JenciL;Davis,ChristieA;Quintero,OmarA
通讯作者：
Quintero,OmarA

Genome-Wide CRISPR Screens Reveal ZATT as a Synthetic Lethal Target of TOP2-Poison Etoposide That Can Act in a TDP2-Independent Pathway.

DOI：
10.3390/ijms24076545
发表时间：
2023-03-31
期刊：
INTERNATIONAL JOURNAL OF MOLECULAR SCIENCES
影响因子：
5.6
作者：
Park, Jeong-Min;Zhang, Huimin;Nie, Litong;Wang, Chao;Huang, Min;Feng, Xu;Tang, Mengfan;Chen, Zhen;Xiong, Yun;Lee, Namsoo;Li, Siting;Yin, Ling;Hart, Traver;Chen, Junjie
通讯作者：
Chen, Junjie

ROCK inhibition abolishes the establishment of the aquiferous system in Ephydatia muelleri (Porifera, Demospongiae).

ROCK 抑制作用消除了 Ephydatia muelleri（Porifera、Demospongiae）中含水系统的建立。

DOI：
10.1016/j.ydbio.2016.02.026
发表时间：
2016
期刊：
Developmental biology
影响因子：
2.7
作者：
Schenkelaars,Quentin;Quintero,Omar;Hall,Chelsea;Fierro-Constain,Laura;Renard,Emmanuelle;Borchiellini,Carole;Hill,AprilL
通讯作者：
Hill,AprilL

Myo19 ensures symmetric partitioning of mitochondria and coupling of mitochondrial segregation to cell division.

Myo19 确保线粒体的对称分配以及线粒体分离与细胞分裂的耦合。

DOI：
10.1016/j.cub.2014.09.045
发表时间：
2014
期刊：
Current biology : CB
影响因子：
0
作者：
Rohn,JenniferL;Patel,JignaV;Neumann,Beate;Bulkescher,Jutta;Mchedlishvili,Nunu;McMullan,RachelC;Quintero,OmarA;Ellenberg,Jan;Baum,Buzz
通讯作者：
Baum,Buzz