Myosin Hinge Region In Contraction

收缩中的肌球蛋白铰链区

• 批准号: 6878060
• 负责人: Sanford I Bernstein
• 金额: $ 35.83万
• 依托单位: SAN DIEGO STATE UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 1996
• 资助国家: 美国
• 起止时间: 1996-04-01 至 2008-03-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/6878060
• 关键词: Drosophilidae; electron microscopy; flying; genetically modified animals; larva; muscle contraction; mutant; myofibrils; myosins; point mutation; protein isoforms; protein structure function; sarcomeres

EXCEED THE SPACE PROVIDED. We propose a multifaceted and integrative approach to determine the function of the "hinge" region of the myosin heavy chain protein in muscle contraction. While much effort has focused on the role of the myosin head in dictating muscle contractile properties, our novel results indicate that the hinge of the rod also is critical for muscle function. We employ Drosophila melanogaster, which is amenable to genetic, transgenic and muscle mechanical approaches. Its single myosin gene encodes alternative myosin hinge regions, one expressed in slow twitch embryonic muscles and one expressed in fast twitch and oscillatory adult muscles. We produced a transgenic line that expresses myosin with the embryonic hinge in muscles that normally express the adult hinge (indirect flight and jump muscles). Transgenic muscles assemble normal-looking myofibrils, but muscle function is severely compromised. We propose to determine if hinge function is critical at the level of the intact organism, the isolated fiber, the single myofibril, the thick filament and/or the myosin molecule, by testing the following hypotheses: 1) the myosin hinge is not important in myofibril assembly; 2) the hinge influences the mechanical properties of muscle fibers and myofibrils [in collaboration with Dr. David Maughan (U. Vermont) and Dr. Gerald Pollack (U. Washington),experts in biophysical measurements of muscle fibers and myofibrils, respectively]; 3) the hinge contributes significantly to physical properties of myosin molecules and thick filaments, specifically to differences in the shortening of isolated myosin molecules or elasticity of intact thick filaments (in collaboration with Dr. Pollack); 4) the propensity to form a coiled-coil is critical to defining the differences between alternative hinge domains; 5) hinge-specific protein interactions impart functional differences between alternativehinge regions [we will test for interaction with the thick filament protein flightin (in collaboration with Dr. Jim Vigoreaux, U. Vermont) and perform genetic suppression studies in flies]. Out integrative approach should elucidate the role of the myosin hinge in muscle function and the mechanism by which it acts. Our work is relevant to human disease since mutations in myosin can cause hypertrophic cardiomyopathy in heart muscle and central core disease in skeletal muscle. Further, since the indirect flight muscle has oscillatory and stretch-activated properties similar to human cardiac muscle, understanding myosin' involvement in generating these properties may lead to insights into human heart function. PERFORMANCE SITE ========================================Section End===========================================

超出所提供的空间。我们提出了一个多方面的和综合的方法来确定肌球蛋白重链蛋白的“铰链”区域在肌肉收缩中的功能。虽然很多努力都集中在肌球蛋白头在支配肌肉收缩性能的作用，我们的新结果表明，铰链杆也是肌肉功能的关键。我们采用果蝇，这是服从遗传，转基因和肌肉机械的方法。它的单个肌球蛋白基因编码替代肌球蛋白铰链区，一个在慢收缩胚胎肌肉中表达，一个在快收缩和振荡成人肌肉中表达。我们生产了一个转基因品系，表达肌球蛋白与胚胎铰链的肌肉，通常表达成人铰链（间接飞行和跳跃肌肉）。转基因肌肉组装正常的肌原纤维，但肌肉功能严重受损。我们建议通过测试以下假设来确定铰链功能在完整生物体、分离的纤维、单个肌原纤维、粗丝和/或肌球蛋白分子的水平上是否是关键的：1）肌球蛋白铰链在肌原纤维组装中不重要; 2）铰链影响肌纤维和肌原纤维的机械性质[与大卫莫恩博士（U.佛蒙特州）和博士杰拉尔德波拉克（美国。华盛顿），分别是肌纤维和肌原纤维的生物物理测量专家]; 3）铰链对肌球蛋白分子和粗肌丝的物理性质有显著贡献，特别是对分离的肌球蛋白分子的缩短或完整的粗肌丝的弹性的差异（与Pollack博士合作）; 4）形成卷曲螺旋的倾向对于定义替代铰链结构域之间的差异至关重要; 5）铰链特异性蛋白质相互作用赋予了不同铰链区之间的功能差异[我们将测试与粗丝蛋白flightin的相互作用（与Jim Vigoreaux博士合作，U。Vermont），并在果蝇中进行遗传抑制研究]。我们的综合方法应该阐明肌球蛋白铰链在肌肉功能中的作用及其作用机制。我们的工作与人类疾病有关，因为肌球蛋白的突变可以导致心肌肥厚性心肌病和骨骼肌中央核心疾病。此外，由于间接飞行肌具有类似于人类心肌的振荡和拉伸激活特性，因此了解肌球蛋白参与产生这些特性可能会导致对人类心脏功能的深入了解。 性能现场=

项目成果

Passive stiffness of Drosophila IFM myofibrils: a novel, high accuracy measurement method.

果蝇 IFM 肌原纤维的被动刚度：一种新颖的高精度测量方法。

• DOI: 10.1007/s10974-004-0684-5
• 发表时间: 2004
• 期刊: Journal of muscle research and cell motility
• 影响因子: 2.7
• 作者: Hao,Yudong;Bernstein,SanfordI;Pollack,GeraldH
• 通讯作者: Pollack,GeraldH

X-ray diffraction from flight muscle with a headless myosin mutation: implications for interpreting reflection patterns.

• DOI: 10.3389/fphys.2014.00416
• 发表时间: 2014
• 期刊: Frontiers in physiology
• 影响因子: 4
• 作者: Iwamoto H;Trombitás K;Yagi N;Suggs JA;Bernstein SI
• 通讯作者: Bernstein SI

Spatially and temporally regulated expression of myosin heavy chain alternative exons during Drosophila embryogenesis.

果蝇胚胎发生过程中肌球蛋白重链替代外显子的空间和时间调节表达。

• DOI: 10.1016/s0925-4773(00)00549-9
• 发表时间: 2001
• 期刊: Mechanisms of development
• 影响因子: 2.6
• 作者: Zhang,S;Bernstein,SI
• 通讯作者: Bernstein,SI

Passive stiffness in Drosophila indirect flight muscle reduced by disrupting paramyosin phosphorylation, but not by embryonic myosin S2 hinge substitution.

• DOI: 10.1529/biophysj.106.088492
• 发表时间: 2006-12
• 期刊: Biophysical journal
• 影响因子: 3.4
• 作者: Yudong Hao;Mark S. Miller;D. Swank;Hongjun Liu;S. Bernstein;D. Maughan;G. Pollack

Drosophila paramyosin is important for myoblast fusion and essential for myofibril formation.

果蝇帕莫辛对于肌细胞融合很重要，对于肌膜纤维的形成至关重要。

• DOI: 10.1083/jcb.200208180
• 发表时间: 2003-03-17
• 期刊: JOURNAL OF CELL BIOLOGY
• 影响因子: 7.8
• 作者: Liu, Hongjun;Mardahl-Dumesnil, Michelle;Sweeney, Sean T;O'Kane, Cahir J;Bernstein, Sanford I
• 通讯作者: Bernstein, Sanford I