Regulation of Microtubule Function by Tubulin Carboxy-terminal Tails

微管蛋白羧基末端尾部对微管功能的调节

基本信息

批准号：
9313705
负责人：
Jeffrey Kyle Moore
金额：
$ 30.15万
依托单位：
UNIVERSITY OF COLORADO DENVER
依托单位国家：
美国
项目类别：
财政年份：
2015
资助国家：
美国
起止时间：
2015-08-01 至 2020-07-31
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/9313705
关键词：
Address Affect Alleles Amino Acids Axonal Transport Behavior Biochemistry Biological Assay Carrier Proteins Cells Charge Chromosomes Complex Defect Disease Electrons Electrostatics Exhibits Future GTP Binding Genetic Genetic Epistasis Genetic Models Goals Hydrolysis In Vitro Investigation Kinesin Laboratories Lead Length Link Malignant Neoplasms Measures Microtubules Mitotic spindle Modeling Molecular Motor Motor Activity Mutation Neuropathy Pathology Pathway interactions Play Polymers Post-Translational Modification Site Post-Translational Protein Processing Preparation Process Property Proteins Regulation Role Signal Transduction Site Surface System Systems Analysis Tail Testing Tissues Tomogram Tubulin Work base cell motility crosslink daughter cell electron tomography experimental study human disease improved in vitro Assay in vivo insight live cell microscopy mutant novel novel strategies protein crosslink protein transport public health relevance tool

项目摘要

DESCRIPTION (provided by applicant): The goal of this project is to understand how microtubule networks are regulated by molecular diversity at the level of tubulin proteins. Tubulin proteins assemble into dynamic microtubule polymers that form networks with associated motor proteins to organize the cell in diverse contexts. Defects in microtubule function are linked to numerous human diseases, including neuropathies and cancer. Understanding the mechanisms that regulate microtubule networks is therefore critical for understanding how defects contribute to the disease state. A major deficit in our understanding is how molecular diversity at the level of tubulin proteins -- mutations, isotype expression, posttranslational modifications -- lead to changes at the level of the network. My laboratory is now poised to address this deficit by overcoming two historical barriers in the microtubule field. First, we have developed a genetic model for manipulating tubulin proteins by directed mutation. Second, we extend our genetic model to complementary systems for analyzing how CTTs affect tubulin biochemistry, MT motor activity, and MT function in vivo. By applying these synergistic approaches, we can distinguish effects on intrinsic microtubule dynamics from effects on motors and other extrinsic regulators, and we can determine the consequences of these effects for a cellular network. In this project, we will determine the roles of carboxy-terminal tail motifs (CTTs) of - and  tubulin. CTTs support electrostatic interactions with motors and other proteins at the MT surface, exhibit sequence diversity across tissue-specific tubulin isotypes, and are major sites of post-translational modification. We hypothesize that CTTs regulate network behavior by promoting microtubule dynamics and the activity of evolutionarily distinct classes of MT motors. We will test this hypothesis in three aims: 1) Determine how CTTs directly affect the assembly of tubulin proteins into microtubule polymers. 2) Determine how CTTs regulate evolutionarily conserved kinesin motors. 3) Define the molecular roles of CTTs in the assembly of a complex cellular microtubule network - the mitotic spindle. The results of this work will provide new insight into the molecular regulation of microtubule networks. Furthermore, these studies will set the stage for future investigations of the roles of tubulin posttranslational modifications, isotyp expression, and disease-associated alleles.

描述（由申请人提供）：该项目的目标是了解微管蛋白水平上的分子多样性如何调节微管网络，微管蛋白组装成动态微管聚合物，与相关运动蛋白形成网络，从而以不同的方式组织细胞。因此，微管功能缺陷与许多人类疾病有关，包括神经病和癌症。因此，了解微管网络的调节机制对于理解缺陷如何导致疾病状态至关重要。我们的理解是微管蛋白水平的分子多样性如何—— 突变、同种型表达、翻译后修饰——导致网络水平的变化。我的实验室现在准备通过克服微管领域的两个历史障碍来解决这一缺陷，首先，我们开发了一种通过定向突变操纵微管蛋白的遗传模型，其次，我们将我们的遗传模型扩展到互补系统，以分析 CTT 如何影响。通过应用这些协同方法，我们可以区分对内在微管动力学的影响与对运动和其他外在调节因子的影响，并确定微管蛋白生物化学、MT 运动活性和 MT 体内功能。这些影响对蜂窝网络的影响。在这个项目中，我们将确定 - 和  的羧基末端尾基序 (CTT) 的作用 CTT 支持与 MT 表面的马达和其他蛋白质的静电相互作用，在组织特异性微管蛋白同种型中表现出序列多样性，并且是 MT 表面的主要位点。我们率先提出，CTT 通过促进微管动力学和进化上不同类别的 MT 马达的活动来调节网络行为。我们将在三个目标中检验这一假设：1）确定 CTT 如何直接影响微管蛋白组装成微管。 2) 确定 CTT 如何调节进化保守的驱动蛋白马达 3) 定义 CTT 在复杂细胞微管网络组装中的分子作用。此外，这些研究将为未来研究微管蛋白翻译后修饰、同型表达和疾病相关等位基因的作用奠定基础。