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MBC in Press, published online ahead of print June 21, 2006
Mol. Biol. Cell 10.1091/mbc.E06-02-0144

A more recent version of this article appeared on September 1, 2006
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Submitted on February 17, 2006
Revised on May 11, 2006
Accepted on June 9, 2006

Caenorhabditis elegans UNC-96 Is a New Component of M-Lines That Interacts with UNC-98 and Paramyosin and Is Required in Adult Muscle for Assembly and/or Maintenance of Thick Filaments

Kristina B. Mercer,* Rachel K. Miller,*{dagger} Tina L. Tinley,*{dagger} Seema Sheth,* Hiroshi Qadota,* and Guy M. Benian*

*Department of Pathology and {dagger}Graduate Division of Biological and Biomedical Sciences, Emory University, Atlanta, GA 30322

Monitoring Editor: Thomas Pollard

To gain further insight into the molecular architecture, assembly and maintenance of the sarcomere, we have carried out a molecular analysis of the UNC-96 protein in the muscle of C. elegans. By polarized light microscopy of bodywall muscle, unc-96 mutants display reduced myofibrillar organization and characteristic birefringent "needles". By immunofluorescent staining of known myofibril components, unc-96 mutants show major defects in the organization of M-lines, and in the localization of a major thick filament component, paramyosin. In unc-96 mutants, the birefringent needles, which contain both UNC-98 and paramyosin, can be suppressed by starvation or by exposure to reduced temperature. UNC-96 is a novel ~47 kDa polypeptide that has regions of similarity to several vertebrate proteins, but no recognizable domains. Antibodies generated to UNC-96 localize the protein to the M-line, a region of the sarcomere in which thick filaments are cross-linked. By genetic and biochemical criteria, UNC-96 interacts with UNC-98, a previously described component of M-lines, and paramyosin. Additionally, UNC-96 copurifies with native thick filaments. A model is presented in which UNC-96 is required in adult muscle to promote thick filament assembly and/or maintenance.

Address correspondence to: Guy M. Benian (pathgb{at}emory.edu)




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