期刊
EXPERIMENTAL CELL RESEARCH
卷 314, 期 19, 页码 3519-3530出版社
ELSEVIER INC
DOI: 10.1016/j.yexcr.2008.09.001
关键词
Nebulin; Myofibrillogenesis; Titin; Actin-binding proteins
资金
- NIH-HLB
- AHA-National council
- Ohio Valley Affiliate
- University of Kentucky
Nebulette is a card iac-specific isoform of the giant actin-binding protein nebulin. Nebulette, having a mass of similar to 100 kDa, is only predicted to extend 150 nm from the edge of the Z-lines. Overexpression of the nebulette C-terminal linker and/or SH3 domains in chicken cardiomyocytes results in a loss of endogenous nebulette with a concomitant loss of tropomyosin (TPM) and troponin, as well as a shortening of the thin filaments. These data suggest that nebulette's position in the sarcomere is important for the maintenance of TPM, troponin and thin filament length. To evaluate this hypothesis, N-terminal nested truncations tagged with GFP were expressed in chicken cardiomyocytes and the cells were analyzed for the distribution of myofilament proteins. Minimal effects on the myofilaments were observed with N-terminal deletions of up to 10 modules: however, deletion of 15 modules replicated the phenotype observed with expression of the C-terminal fragments. Expression of internal deletions of nebulette verifies that a site between module 10 and 15 is important for TPM maintenance within the sarcomeric lattice. We have additionally isolated TPM cDNAs from a yeast two hybrid (Y2H) analysis. These data indicate the importance of the nebulette-TPM interactions in the maintenance and stability of the thin filaments. (C) 2008 Elsevier Inc. All rights reserved.
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