Nematode UNC-98 functions in focal adhensions and nuclei

线虫 UNC-98 在粘着点和细胞核中发挥作用

• 批准号: 7483267
• 负责人: GUY Martin BENIAN
• 金额: $ 23.35万
• 依托单位: EMORY UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2005
• 资助国家: 美国
• 起止时间: 2005-09-01 至 2010-08-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7483267
• 关键词: Adhesions; Adult; Affect; Alleles; Animals; Antibodies; Binding; Biochemical Genetics; Caenorhabditis elegans; Cell Nucleus; Class; Data; Development; Embryonic Development; Face; Fingers; Focal Adhesions; Gene Expression; Gene Targeting; Genes; Genetic; Goals; Green Fluorescent Proteins; Growth; Homeostasis; Human; Hybrids; In Vitro; LIM Domain Protein; LIMS1 gene; Life; Localized; Location; Maintenance; Mammals; Messenger RNA; Microfilaments; Molecular; Monitor; Muscle; Muscle Cells; Myocardium; Myofibrils; Myopathy; Nematoda; Nuclear; Numbers; Paralysed; Paratropomyosin; Peptide Signal Sequences; Phenotype; Proteins; Reporting; Sarcomeres; Site; Skeletal system; Structure; Thick Filament; Travel; analog; insight; mutant; novel; polypeptide; promoter; research study

DESCRIPTION (provided by applicant): The long-term goal is to understand the mechanism by which myofibrils assemble from their components, and how these precise structures are maintained in the face of repeated muscle activity. This goal has relevance to many types of human muscle disease, including those of skeletal and cardiac muscle. This proposal outlines studies of three genes in C. elegans, unc-98, unc-96 and unc-97 which are required for proper myofibril assembly and/or maintenance. UNC-98 is a novel 310 residue polypeptide consisting of 4 C2H2 Zn fingers and predicted NLS and NES sequences. By use of UNC-98 antibodies and UNC-98:GFP fusions, UNC-98 resides at M-lines, dense bodies (Z line analogs) and muscle cell nuclei. UNC-98 interacts with UNC-97 (PINCH in mammals), a LIM domain protein required for muscle focal adhesion assembly. Like UNC-98, UNC-97:GFP localizes to dense bodies, M-lines and nuclei. It is hypothesized that UNC-98 and UNC-97 function in muscle focal adhesion homeostasis, in which these proteins when localized to the adhesion sites monitor myofibril structure or activity, and travel to the nucleus to affect gene expression, unc-96 has a similar mutant phenotype to unc-98, interacts with unc-98 genetically, and may protect the sarcomere from breakdown resulting from normal muscle usage. We have determined that UNC-96 is a novel 418 aa protein and by 2 hybrid interacts with two conserved LIM domain proteins that also interact with UNC-97. Goals include: (1) for UNC-98, prove that its nuclear localization is important for its function, show that it can move from myofibrils into the nucleus, determine whether it can influence the expression of other genes, and whether it interacts with paramyosin in thick filaments; (2) for UNC-96, find out where it is localized in the sarcomere, provide further evidence that it interacts with paramyosin, UNC-98 and two LIM domain proteins; (3) for UNC-97 study its dynamics, whether it influences gene expression, whether its nuclear localization depends on UNC-98, and whether it indeed interacts with the two LIM domain proteins.

描述（由申请人提供）：长期目标是了解肌原纤维从其组件中组装的机制，以及面对重复的肌肉活动时如何保持这些精确的结构。该目标与许多类型的人类肌肉疾病有关，包括骨骼和心脏肌肉。该提案概述了秀丽隐杆线虫，UNC-98，UNC-96和UNC-97中三个基因的研究，这些研究是适当的肌原纤维组装和/或维护所必需的。 UNC-98是一种新型的310个残基多肽，由4个C2H2 Zn手指和预测的NLS和NES序列组成。通过使用UNC-98抗体和UNC-98：GFP融合，UNC-98驻留在M线上，密集的身体（Z线模拟）和肌肉细胞核。 UNC-98与UNC-97（哺乳动物中的捏）相互作用，这是肌肉局灶性粘附组件所需的LIM结构蛋白。像UNC-98一样，UNC-97：GFP定位于密集的身体，M线和核。据推测，UNC-98和UNC-97在肌肉局灶性稳态中的功能，其中这些蛋白质本地定位于粘附部位监测肌原纤维的结构或活动，并前往细胞核以影响基因表达，UNC-96具有与UNC-98相似的突变表型，与UNC-98相似，从而使UNC-98的群体相互作用，并可以在sARCEN上与SARCEN相互作用。我们已经确定UNC-96是一种新型的418 AA蛋白，通过2杂交与两个保守的LIM结构蛋白相互作用，它们也与UNC-97相互作用。目标包括：（1）对于UNC-98，证明其核定位对其功能很重要，表明它可以从肌原纤维转移到核中，确定它是否可以影响其他基因的表达，以及它是否在厚细丝中与帕诺蛋白相互作用。 （2）对于UNC-96，找出它位于肌节中的位置，提供了进一步的证据，表明它与paramyosin，UNC-98和两个LIM结构域蛋白相互作用； （3）对于UNC-97研究其动力学，它是否影响基因表达，其核定位是否取决于UNC-98，以及它是否确实与两个LIM结构域蛋白相互作用。