Dissection of the Salm Mediated Transcriptional Switch to Fibrillar Myogenesis

Salm 介导的转录转换到纤维肌生成的剖析

• 批准号: 8476153
• 负责人: Maria L Spletter
• 金额: $ 4.61万
• 依托单位: MAX PLANCK INSTITUTE OF BIOCHEMISTRY
• 依托单位国家: 美国
• 财政年份: 2012
• 资助国家: 美国
• 起止时间: 2012-05-01 至 2014-04-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8476153
• 关键词: Adult; Aging; Alternative Splicing; Binding; Biology; Cardiac Myocytes; ChIP-seq; Congenital Heart Defects; Consensus; Data; Defect; Development; Disease; Dissection; Drosophila genus; Event; Exhibits; Generations; Genes; Goals; Histocompatibility Testing; Human; Jellyfish; Knowledge; Life; Logic; Mediating; Metabolic; Molecular; Molecular Profiling; Muscle; Muscle Development; Muscle Fibers; Muscle Weakness; Muscle function; Muscular Dystrophies; Myoblasts; Myocardium; Network-based; Phenotype; Physiological Adaptation; Play; Process; Protein Isoforms; RNA Interference; RNA Splicing; Regulation; Role; Sampling; Sarcomeres; Series; Signal Transduction; Specific qualifier value; Spliced Genes; Staging; Stem cells; Striated Muscles; System; Tendon structure; Tissue-Specific Gene Expression; Tissues; Tube; Tubular formation; Vertebrates; age related; base; chromatin immunoprecipitation; developmental plasticity; fibrillogenesis; fly; insight; migration; mutant; myogenesis; novel; programs; transcription factor; transcriptome sequencing

DESCRIPTION (provided by applicant): Muscle development proceeds through an evolutionarily conserved series of events, including myoblast generation and migration, myotube formation and attachment to tendons, sarcomerogenesis and myofiber maturation. In the course of this process, stem-cell-like muscle precursors differentiate forming syncytial tubes with a specialized contractile apparatus. Drosophila adult muscles, which exhibit developmental plasticity and functional diversification similar to that observed in vertebrates, include fibrilla and tubular types. Tubular muscles are similar to vertebrate striated muscle, while fibrilar indirect flight muscles (IFMs) share some features with cardiomyocytes. Defects in muscle development, and possibly in the specification of muscle types, can result in disorders, for example muscular dystrophies or congenital heart defects, which result in debilitating and life-threatening conditions. Recently, Spalt major (Salm) was characterized as a master regulator both necessary and sufficient to mediate the switch to the fibrillar muscle identity in Drosophila. However, the detailed mechanisms of fibrillar muscle construction remain unclear. Here I propose to combine RNA-Seq with ChIP-Seq to characterize the Salm mediated transcriptional switch and identify Salm effectors that execute fibrillogenesis. I will perform a developmental RNA-Seq timecourse to characterize dynamic expression changes during flight-muscle development and perform ChIP-Seq to identify direct Salm targets. As alternative splicing plays a major role in myogenesis and alternatively spliced gene isoforms are misexpressed in Salm RNAi flies, I hypothesized that Salm regulates differential splicing. I propose to investigate the splicing factor Arrest to gain mechanistic insight into how Salm induced alternative splicing may dictate the development of fibrillar tissue. Dissection of the Salm mediated transcriptional switch and its molecular effectors will elucidate mechanisms of sarcomerogenesis that should be relevant to vertebrate myogenesis.

描述（由申请人提供）：肌肉发育是通过进化保守的事件进行的，包括成肌细胞产生和迁移，肌管形成以及对肌腱的附着，肉瘤发生和肌纤维成熟。在此过程的过程中，干细胞样的肌肉前体与专门的收缩装置区分形成合成管。果蝇成年肌肉表现出与脊椎动物中观察到的相似的发育可塑性和功能多样化，包括原纤维和管状类型。管状肌肉类似于脊椎动物横纹的肌肉，而纤维间的间接飞行肌肉（IFM）与心肌细胞共享某些特征。肌肉发育的缺陷以及肌肉类型的规范可能会导致疾病，例如肌肉营养不良或先天性心脏缺陷，从而导致衰弱和威胁生命的疾病。最近，Spalt Major（SALM）被描述为主要调节剂，足以介导果蝇中的纤维肌身份的转换。 但是，纤维肌构建的详细机制尚不清楚。在这里，我建议将RNA-Seq与ChIP-Seq相结合，以表征SALM介导的转录开关，并识别执行原纤维生成的SALM效应子。我将执行一个发育性RNA-seq时期，以表征飞行肌肉发育过程中动态表达变化并执行芯片seq以识别直接SALM目标。由于替代剪接在肌发生中起主要作用，而剪接的基因同工型在Salm RNAi Flies中被压抑，因此我假设SALM调节差异剪接。我建议研究剪接因子停滞，以获取机械洞察力，以了解Salm诱导的替代剪接如何决定原纤维组织的发展。 SALM介导的转录开关的解剖 它的分子效应子将阐明应与脊椎动物肌发生有关的肉瘤发生机制。