Beyond the tubulin code: Understanding how subunit diversity regulates the formation and function of microtubules

超越微管蛋白代码：了解亚基多样性如何调节微管的形成和功能

• 批准号: 10164813
• 负责人: Jeffrey Kyle Moore
• 金额: $ 37.62万
• 依托单位: UNIVERSITY OF COLORADO DENVER
• 依托单位国家: 美国
• 财政年份: 2020
• 资助国家: 美国
• 起止时间: 2020-05-15 至 2025-04-30
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10164813
• 关键词: Amino Acid Sequence; Behavior; Binding Proteins; Buffers; Cations; Cell physiology; Cells; Cellular biology; Code; Complex; Developmental Process; Disease; Environment; Genetic; Genetic Models; Goals; Heterogeneity; Kinesin; Knowledge; Lead; Malignant Neoplasms; Microtubules; Modeling; Molecular; Molecular Conformation; Motor; Neuropathy; Play; Post-Translational Protein Processing; Protein Biochemistry; Proteins; Role; Set protein; Structure; Surface; Tail; Tissues; Tubulin; cell motility; human disease; insight; live cell imaging; novel

Project Summary A fundamental question in cell biology is how cells build functionally and structurally distinct microtubule (MT) networks using a seemingly simple set of protein building blocks -- -tubulin heterodimers. For many years, the cytoskeletal field has focused on the roles of MT-binding proteins and motors as the primary regulators of network structure and function. However, it is now clear that the -tubulin building blocks are not so simple. Rather than a uniform track, the MT surface is a molecularly diverse landscape that is generated by genetic and posttranslational differences between -tubulins. The `tubulin code' model posits that changes to the intrinsically disordered carboxy-terminal tail (CTT) domains of -tubulins create a molecular code at the MT surface that is “read” by MT-binding proteins. The overarching goals of this proposal are to establish mechanistic connections between CTTs and the conformational diversity of MT ends and lattices, and to understand how this regulates complex MT network functions at the cellular level. This structure proposal features and function, and a multi-system, multi-scale approach to understanding how how these lead to changes in the function of MT networks CTTs impact tubulin in cells. My lab has established expertise in investigating tubulin using approaches that integrate genetic models, live-cell imaging and protein biochemistry. Our progress over the past five years has furthered our understanding of how - tubulin CTTs regulate MT networks, and, more broadly, how tubulin heterogeneity impacts cellular and developmental processes. The proposed project will build upon our expertise to expand the current model of the tubulin code and give broader insights into mechanisms of MT function. Our goals for the next five years include 1) define how CTTs guide the structure of MT ends to regulate MT dynamics, 2) determine how blends of -tubulins with different amino acid sequences and posttranslational modifications give rise to complex behaviors at the level of MT networks in cells, 3) define how CTTs promote the directionality of kinesin motility along MTs, and 4) establish a novel role for tubulins in buffering intracellular cation concentrations. Our synergistic approach is uniquely suited to advance knowledge of tubulin structure and function that will be important in a broad range of contexts, provide new insights into how microtubule networks regulate and respond to changes at the level of tubulin subunits, and how these impact different cellular contexts.

项目摘要 细胞生物学中的一个基本问题是细胞在功能和结构上不同的微管（MT）上的构建方式 使用看似简单的蛋白质构建块的网络 - 微管蛋白异二聚体。多年， 细胞骨架场的重点是MT结合蛋白和电机作为主要调节剂的作用 网络结构和功能。但是，现在很明显，微管蛋白的构建块并不那么简单。 MT表面不是统一的轨道，是一种分子多样的景观，由遗传产生 微管蛋白之间的翻译后差异。 “微管蛋白代码”模型认为变为 微管蛋白的本质上受干扰的羧基末端尾巴（CTT）结构域在MT上产生分子代码 通过MT结合蛋白“读取”的表面。该提议的总体目标是建立 CTT和MT末端和格子的会议多样性之间的机械连接，以及 了解该如何调节复杂的MT网络在细胞水平上的功能。 这 结构 提案功能 和功能，以及 一种多系统的多尺度方法，以理解如何 这些如何导致MT网络的功能变化 CTT 影响 微管蛋白 在 细胞。 我的实验室有 使用整合遗传模型的方法，实时细胞成像来研究小管蛋白方面的专业知识 和蛋白质生物化学。在过去五年中，我们的进步已经进一步发展了我们对-的理解 软骨素CTTS调节MT网络，更广泛地，小节肽异质性如何影响细胞和 发展过程。拟议的项目将在我们的专家基础上建立，以扩大当前的模型 小管蛋白代码并对MT功能机制提供更广泛的见解。我们未来五年的目标 包括1）定义CTT如何指导MT末端的结构以调节MT动力学，2）确定如何混合 具有不同氨基酸序列和翻译后修饰的微管蛋白会导致复杂 细胞中MT网络级别的行为，3）定义CTT如何促进驱动蛋白运动的方向性 沿着MTS和4）在缓冲细胞内阳离子浓度中建立了小管蛋白的新作用。我们的 协同方法非常适合促进对小管蛋白结构和功能的了解 在各种环境中重要的重要性，提供有关微管网络如何调节和的新见解 响应微管蛋白亚基水平的变化，以及它们如何影响不同的细胞环境。