The UNC-45 Chaperone as a Modulator of Myosin Biogenesis and Function

UNC-45 伴侣作为肌球蛋白生物合成和功能的调节剂

基本信息

批准号：
9789043
负责人：
GUY Martin BENIAN
金额：
$ 30.69万
依托单位：
UNIVERSITY OF TEXAS MED BR GALVESTON
依托单位国家：
美国
项目类别：
财政年份：
2016
资助国家：
美国
起止时间：
2016-05-01 至 2022-04-30
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/9789043
关键词：
Address Amino Acids Animals Binding Biochemical Biogenesis Biological Biological Assay Biological Models Biology Biophysics CRISPR/Cas technology Caenorhabditis elegans Cardiomyopathies Cell division Clustered Regularly Interspaced Short Palindromic Repeats Collaborations Data Development Drosophila genus Equilibrium Exposure to Functional disorder Genes Head Heat Stress Disorders Heat-Shock Proteins 90 Heat-Shock Response Hydrogen Peroxide In Vitro Investigation Knowledge Lead Length Mediating Microfilaments Molecular Molecular Chaperones Molecular Conformation Motor Muscle Muscle Cells Muscle Development Muscle function Mutagenesis Mutation Myofibrils Myopathy Myosin ATPase Myosin Type II Phenotype Physiological Process Protein Isoforms Proteins Publishing Recovery Regulation Research Research Personnel Role Sarcomeres Series Site Skeletal Muscle Stress Striated Muscles Structure System Techniques Testing Thick Filament Up-Regulation Vertebrates Work cell motility cell type crystallinity developmental disease experimental study human disease in vivo inhibitor/antagonist insight member multidisciplinary mutant non-muscle myosin novel novel therapeutic intervention novel therapeutics premature prevent protein complex protein folding public health relevance recruit repaired response skeletal stem temperature sensitive mutant tumor tumor progression

项目摘要

DESCRIPTION (provided by applicant): In this proposal we aim to elucidate the precise molecular mechanisms by which the chaperones UNC-45 and Hsp90 assist in the folding of the myosin head which is critical for sarcomere assembly during development and repair of stress-induced damage to sarcomeres in mature muscle. Developing an understanding of these mechanisms is a problem at the core of muscle development and function. It is also highly relevant to the folding and repair of the many classes of myosin molecules that are expressed in non-muscle tissues. Our recent discovery that myosin attains a novel conformation, which hydrolyzes ATP at the normal rate but with a greatly reduced ability to translocate actin filaments, in the presence of UNC-45 and is reversed by Hsp90, raises important questions as to the exact roles of these chaperones in myosin folding and repair and myofilament assembly. Guided by our recent published and preliminary studies, we hypothesize that during myosin assembly or repair, UNC-45 assists the myosin head to attain its native conformation and subsequently locks it in a state that prevents premature powerstrokes; this state is relieved by Hsp90 upon successful assembly into the sarcomere or repair of myosin heads. This hypothesis will be tested by using a combination of in vitro and in vivo analyses through a unique collaboration between two expert research labs. In Aim 1 we will use mutagenesis, biochemical and biophysical assays, including myosin-dependent actin filament gliding assays, to establish the UNC-45 domains and key amino acid regions that mediate the different unique functions of UNC-45. In Aims 2 and 3 we will use the power of C. elegans as a model system to examine the functional relationships between mutant UNC-45 proteins and sarcomere assembly (Aim 2), and the dynamics of association of UNC-45 and Hsp90 with themselves and with myosin during repair of damaged myosin heads (Aim 3). Our hypothesis, if confirmed, will represent a new paradigm in the biology of myosins, with potential novel therapeutic approaches not only for striated muscle disorders stemming from mutations in sarcomeric proteins (skeletal myopathies and cardiomyopathies), but also for the problem of tumor invasiveness.

描述（由适用提供）：在此提案中，我们旨在阐明链酮UNC-45和HSP90的精确分子机制，有助于肌球蛋白头的折叠，这对于肌动蛋白组装至关重要，这对于肌动蛋白组装至关重要。在肌肉发育和功能的核心上发展对这些机制是一个问题。它也与在非肌肉时机中表达的许多类别的肌球蛋白分子的折叠和修复高度相关。我们最近发现，肌球蛋白达到了一种新型的构象，该构象以正常速率水解ATP，但在UNC-45的存在下，肌球蛋白具有正常的构象，但其能力大大降低了易位的肌动蛋白丝的能力，并由HSP90逆转，提出了有关这些斑点在肌动蛋白折叠和修复和修复和肌纤维组合中的精确作用的重要问题。在我们最近发表和初步研究的指导下，我们假设在肌球蛋白组装或修复过程中，UNC-45有助于肌球蛋白负责人获得其本地会议，然后将其锁定在阻止过早的PowerStrokes的状态下。成功组装后，HSP90救出了该状态，以肌球蛋白头的修复。该假设将通过在两个专家研究实验室之间的独特协作中使用体外和体内分析的组合来检验。在AIM 1中，我们将使用诱变，生化和生物物理测定，包括肌球蛋白依赖性肌动蛋白丝滑行测定，以建立介导UNC-45的不同独特功能的UNC-45结构域和关键氨基酸区域。在目标2和3中，我们将使用秀丽隐杆线虫作为模型系统来检查突变体UNC-45蛋白质和肌膜组装之间的功能关系（AIM 2），以及在修复受损的肌球蛋白头（AIM 3）期间，UNC-45和HSP90与自己以及肌球蛋白与肌球蛋白的关联动力学（AIM 3）。如果得到证实，我们的假设将代表肌球蛋白生物学生物学的新范式，不仅对肌肉蛋白突变（骨骼肌病和心肌病）突变引起的潜在的新型热方法，而且还出于肿瘤浸润性的问题。

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）

Meeting report – NSF-sponsored workshop ‘Progress and Prospects of Single-Molecule Force Spectroscopy in Biological and Chemical Sciences’

会议报告 — NSF 主办的研讨会 — 单分子力谱在生物和化学科学领域的进展与展望 —

DOI：
10.1242/jcs.251421
发表时间：
2020
期刊：
Journal of Cell Science
影响因子：
4
作者：
Marszalek, Piotr E.;Oberhauser, Andres F.
通讯作者：
Oberhauser, Andres F.

Mutational Analysis of the Structure and Function of the Chaperoning Domain of UNC-45B.

UNC-45B 陪伴结构域的结构和功能的突变分析。

DOI：
10.1016/j.bpj.2020.07.012
发表时间：
2020
期刊：
Biophysical journal
影响因子：
3.4
作者：
Gaziova,Ivana;Moncrief,Taylor;Christian,CourtneyJ;Villarreal,Michael;Powell,Simon;Lee,Hubert;Qadota,Hiroshi;White,MarkA;Benian,GuyM;Oberhauser,AndresF
通讯作者：
Oberhauser,AndresF

The central domain of UNC-45 chaperone inhibits the myosin power stroke.

DOI：
10.1002/2211-5463.12346
发表时间：
2018-01
期刊：
FEBS open bio
影响因子：
2.6
作者：
Bujalowski PJ;Nicholls P;Garza E;Oberhauser AF
通讯作者：
Oberhauser AF

Mutations in conserved residues of the myosin chaperone UNC-45 result in both reduced stability and chaperoning activity.

肌球蛋白伴侣 UNC-45 保守残基的突变会导致稳定性和伴侣活性降低。

DOI：
10.1002/pro.4180
发表时间：
2021
期刊：
Protein science : a publication of the Protein Society
影响因子：
0
作者：
Moncrief,Taylor;Matheny,CourtneyJ;Gaziova,Ivana;Miller,JohnM;Qadota,Hiroshi;Benian,GuyM;Oberhauser,AndresF
通讯作者：
Oberhauser,AndresF

Animal models of sarcopenia.