Vinculin is essential for muscle function in the nematode.
Actin filaments in the body wall muscle of the nematode Caenorhabditis elegans are attached to the sarcolemma through vinculin-containing structures called dense bodies, Z-line analogues. To investigate the in vivo function of vinculin, we executed a genetic screen designed to recover mutations in the region of the nematode vinculin gene, deb-1. According to four independent criteria, two of the isolated mutants were shown to be due to alterations in the deb-1 gene. First, antibody staining showed that the mutants had reduced levels of vinculin. Second, the sequence of each mutant gene was altered from that of wild type, with one mutation altering a conserved splice sequence and the other generating a premature amber stop codon. Third, the amber mutant was suppressed by the sup-7 amber suppressor tRNA gene. Finally, injection of a cloned wild type copy of the gene rescued the mutant. Mutant animals lacking vinculin arrested development as L1 larvae. In such animals, embryonic elongation was interrupted at the twofold length, so that the mutants were shorter than wild type animals at the same stage. The mutants were paralyzed and had disorganized muscle, a phenotype consistent with the idea that vinculin is essential for muscle function in the nematode.
Barstead
R J
RJ
Washington University School of Medicine, Department of Genetics, St. Louis, Missouri 63110.
Waterston
R H
RH
D000818:Animals; D002107:Caenorhabditis / Q000196:embryology / Q000235:genetics / Q000502:physiology*; D002874:Chromosome Mapping; D003433:Crosses, Genetic; D003598:Cytoskeletal Proteins / Q000235:genetics / Q000502:physiology*; D004247:DNA / Q000235:genetics; D004625:Embryo, Nonmammalian / Q000502:physiology; D005260:Female; D005816:Genetic Complementation Test; D008040:Genetic Linkage; D008297:Male; D009132:Muscles / Q000502:physiology*; D009154:Mutation; D009692:Nucleic Acid Heteroduplexes / Q000235:genetics; D014170:Transformation, Genetic; D016596:Vinculin
D003598:Cytoskeletal Proteins; D009692:Nucleic Acid Heteroduplexes; D016596:Vinculin; D004247:DNA