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. 1991 Aug 2;114(4):715–724. doi: 10.1083/jcb.114.4.715

Vinculin is essential for muscle function in the nematode

PMCID: PMC2289884  PMID: 1907975

Abstract

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.

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Selected References

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  1. Anderson R. A., Marchesi V. T. Regulation of the association of membrane skeletal protein 4.1 with glycophorin by a polyphosphoinositide. Nature. 1985 Nov 21;318(6043):295–298. doi: 10.1038/318295a0. [DOI] [PubMed] [Google Scholar]
  2. Antin P. B., Tokunaka S., Nachmias V. T., Holtzer H. Role of stress fiber-like structures in assembling nascent myofibrils in myosheets recovering from exposure to ethyl methanesulfonate. J Cell Biol. 1986 Apr;102(4):1464–1479. doi: 10.1083/jcb.102.4.1464. [DOI] [PMC free article] [PubMed] [Google Scholar]
  3. Ardizzi J. P., Epstein H. F. Immunochemical localization of myosin heavy chain isoforms and paramyosin in developmentally and structurally diverse muscle cell types of the nematode Caenorhabditis elegans. J Cell Biol. 1987 Dec;105(6 Pt 1):2763–2770. doi: 10.1083/jcb.105.6.2763. [DOI] [PMC free article] [PubMed] [Google Scholar]
  4. Barstead R. J., Waterston R. H. The basal component of the nematode dense-body is vinculin. J Biol Chem. 1989 Jun 15;264(17):10177–10185. [PubMed] [Google Scholar]
  5. Bendori R., Salomon D., Geiger B. Identification of two distinct functional domains on vinculin involved in its association with focal contacts. J Cell Biol. 1989 Jun;108(6):2383–2393. doi: 10.1083/jcb.108.6.2383. [DOI] [PMC free article] [PubMed] [Google Scholar]
  6. Branton D., Cohen C. M., Tyler J. Interaction of cytoskeletal proteins on the human erythrocyte membrane. Cell. 1981 Apr;24(1):24–32. doi: 10.1016/0092-8674(81)90497-9. [DOI] [PubMed] [Google Scholar]
  7. Brenner S. The genetics of Caenorhabditis elegans. Genetics. 1974 May;77(1):71–94. doi: 10.1093/genetics/77.1.71. [DOI] [PMC free article] [PubMed] [Google Scholar]
  8. Burridge K., Fath K., Kelly T., Nuckolls G., Turner C. Focal adhesions: transmembrane junctions between the extracellular matrix and the cytoskeleton. Annu Rev Cell Biol. 1988;4:487–525. doi: 10.1146/annurev.cb.04.110188.002415. [DOI] [PubMed] [Google Scholar]
  9. Burridge K., Mangeat P. An interaction between vinculin and talin. Nature. 1984 Apr 19;308(5961):744–746. doi: 10.1038/308744a0. [DOI] [PubMed] [Google Scholar]
  10. Cotton R. G., Rodrigues N. R., Campbell R. D. Reactivity of cytosine and thymine in single-base-pair mismatches with hydroxylamine and osmium tetroxide and its application to the study of mutations. Proc Natl Acad Sci U S A. 1988 Jun;85(12):4397–4401. doi: 10.1073/pnas.85.12.4397. [DOI] [PMC free article] [PubMed] [Google Scholar]
  11. Coulson A., Sulston J., Brenner S., Karn J. Toward a physical map of the genome of the nematode Caenorhabditis elegans. Proc Natl Acad Sci U S A. 1986 Oct;83(20):7821–7825. doi: 10.1073/pnas.83.20.7821. [DOI] [PMC free article] [PubMed] [Google Scholar]
  12. Coulson A., Waterston R., Kiff J., Sulston J., Kohara Y. Genome linking with yeast artificial chromosomes. Nature. 1988 Sep 8;335(6186):184–186. doi: 10.1038/335184a0. [DOI] [PubMed] [Google Scholar]
  13. Coutu M. D., Craig S. W. cDNA-derived sequence of chicken embryo vinculin. Proc Natl Acad Sci U S A. 1988 Nov;85(22):8535–8539. doi: 10.1073/pnas.85.22.8535. [DOI] [PMC free article] [PubMed] [Google Scholar]
  14. Dlugosz A. A., Antin P. B., Nachmias V. T., Holtzer H. The relationship between stress fiber-like structures and nascent myofibrils in cultured cardiac myocytes. J Cell Biol. 1984 Dec;99(6):2268–2278. doi: 10.1083/jcb.99.6.2268. [DOI] [PMC free article] [PubMed] [Google Scholar]
  15. DuBose R. F., Hartl D. L. Rapid purification of PCR products for DNA sequencing using Sepharose CL-6B spin columns. Biotechniques. 1990 Mar;8(3):271–274. [PubMed] [Google Scholar]
  16. Fire A. Integrative transformation of Caenorhabditis elegans. EMBO J. 1986 Oct;5(10):2673–2680. doi: 10.1002/j.1460-2075.1986.tb04550.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  17. Francis G. R., Waterston R. H. Muscle organization in Caenorhabditis elegans: localization of proteins implicated in thin filament attachment and I-band organization. J Cell Biol. 1985 Oct;101(4):1532–1549. doi: 10.1083/jcb.101.4.1532. [DOI] [PMC free article] [PubMed] [Google Scholar]
  18. Geiger B. A 130K protein from chicken gizzard: its localization at the termini of microfilament bundles in cultured chicken cells. Cell. 1979 Sep;18(1):193–205. doi: 10.1016/0092-8674(79)90368-4. [DOI] [PubMed] [Google Scholar]
  19. Ito S., Werth D. K., Richert N. D., Pastan I. Vinculin phosphorylation by the src kinase. Interaction of vinculin with phospholipid vesicles. J Biol Chem. 1983 Dec 10;258(23):14626–14631. [PubMed] [Google Scholar]
  20. Jones P., Jackson P., Price G. J., Patel B., Ohanion V., Lear A. L., Critchley D. R. Identification of a talin binding site in the cytoskeletal protein vinculin. J Cell Biol. 1989 Dec;109(6 Pt 1):2917–2927. doi: 10.1083/jcb.109.6.2917. [DOI] [PMC free article] [PubMed] [Google Scholar]
  21. Kramer J. M., French R. P., Park E. C., Johnson J. J. The Caenorhabditis elegans rol-6 gene, which interacts with the sqt-1 collagen gene to determine organismal morphology, encodes a collagen. Mol Cell Biol. 1990 May;10(5):2081–2089. doi: 10.1128/mcb.10.5.2081. [DOI] [PMC free article] [PubMed] [Google Scholar]
  22. Lin Z. X., Holtzer S., Schultheiss T., Murray J., Masaki T., Fischman D. A., Holtzer H. Polygons and adhesion plaques and the disassembly and assembly of myofibrils in cardiac myocytes. J Cell Biol. 1989 Jun;108(6):2355–2367. doi: 10.1083/jcb.108.6.2355. [DOI] [PMC free article] [PubMed] [Google Scholar]
  23. Milam L. M. Electron microscopy of rotary shadowed vinculin and vinculin complexes. J Mol Biol. 1985 Aug 5;184(3):543–545. doi: 10.1016/0022-2836(85)90301-8. [DOI] [PubMed] [Google Scholar]
  24. Miller D. M., 3rd, Ortiz I., Berliner G. C., Epstein H. F. Differential localization of two myosins within nematode thick filaments. Cell. 1983 Sep;34(2):477–490. doi: 10.1016/0092-8674(83)90381-1. [DOI] [PubMed] [Google Scholar]
  25. Montandon A. J., Green P. M., Giannelli F., Bentley D. R. Direct detection of point mutations by mismatch analysis: application to haemophilia B. Nucleic Acids Res. 1989 May 11;17(9):3347–3358. doi: 10.1093/nar/17.9.3347. [DOI] [PMC free article] [PubMed] [Google Scholar]
  26. Niggli V., Dimitrov D. P., Brunner J., Burger M. M. Interaction of the cytoskeletal component vinculin with bilayer structures analyzed with a photoactivatable phospholipid. J Biol Chem. 1986 May 25;261(15):6912–6918. [PubMed] [Google Scholar]
  27. Otto J. J. Detection of vinculin-binding proteins with an 125I-vinculin gel overlay technique. J Cell Biol. 1983 Oct;97(4):1283–1287. doi: 10.1083/jcb.97.4.1283. [DOI] [PMC free article] [PubMed] [Google Scholar]
  28. Otto J. J. Vinculin. Cell Motil Cytoskeleton. 1990;16(1):1–6. doi: 10.1002/cm.970160102. [DOI] [PubMed] [Google Scholar]
  29. Padgett R. A., Grabowski P. J., Konarska M. M., Seiler S., Sharp P. A. Splicing of messenger RNA precursors. Annu Rev Biochem. 1986;55:1119–1150. doi: 10.1146/annurev.bi.55.070186.005351. [DOI] [PubMed] [Google Scholar]
  30. Price G. J., Jones P., Davison M. D., Patel B., Eperon I. C., Critchley D. R. Isolation and characterization of a vinculin cDNA from chick-embryo fibroblasts. Biochem J. 1987 Jul 15;245(2):595–603. doi: 10.1042/bj2450595. [DOI] [PMC free article] [PubMed] [Google Scholar]
  31. Priess J. R., Hirsh D. I. Caenorhabditis elegans morphogenesis: the role of the cytoskeleton in elongation of the embryo. Dev Biol. 1986 Sep;117(1):156–173. doi: 10.1016/0012-1606(86)90358-1. [DOI] [PubMed] [Google Scholar]
  32. Saiki R. K., Gelfand D. H., Stoffel S., Scharf S. J., Higuchi R., Horn G. T., Mullis K. B., Erlich H. A. Primer-directed enzymatic amplification of DNA with a thermostable DNA polymerase. Science. 1988 Jan 29;239(4839):487–491. doi: 10.1126/science.2448875. [DOI] [PubMed] [Google Scholar]
  33. Schedl T., Kimble J. fog-2, a germ-line-specific sex determination gene required for hermaphrodite spermatogenesis in Caenorhabditis elegans. Genetics. 1988 May;119(1):43–61. doi: 10.1093/genetics/119.1.43. [DOI] [PMC free article] [PubMed] [Google Scholar]
  34. Sulston J. E., Schierenberg E., White J. G., Thomson J. N. The embryonic cell lineage of the nematode Caenorhabditis elegans. Dev Biol. 1983 Nov;100(1):64–119. doi: 10.1016/0012-1606(83)90201-4. [DOI] [PubMed] [Google Scholar]
  35. Wachsstock D. H., Wilkins J. A., Lin S. Specific interaction of vinculin with alpha-actinin. Biochem Biophys Res Commun. 1987 Jul 31;146(2):554–560. doi: 10.1016/0006-291x(87)90564-x. [DOI] [PubMed] [Google Scholar]
  36. Waterston R. H. A second informational suppressor, SUP-7 X, in Caenorhabditis elegans. Genetics. 1981 Feb;97(2):307–325. doi: 10.1093/genetics/97.2.307. [DOI] [PMC free article] [PubMed] [Google Scholar]
  37. Waterston R. H. The minor myosin heavy chain, mhcA, of Caenorhabditis elegans is necessary for the initiation of thick filament assembly. EMBO J. 1989 Nov;8(11):3429–3436. doi: 10.1002/j.1460-2075.1989.tb08507.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  38. Wills N., Gesteland R. F., Karn J., Barnett L., Bolten S., Waterston R. H. The genes sup-7 X and sup-5 III of C. elegans suppress amber nonsense mutations via altered transfer RNA. Cell. 1983 Jun;33(2):575–583. doi: 10.1016/0092-8674(83)90438-5. [DOI] [PubMed] [Google Scholar]
  39. Zengel J. M., Epstein H. F. Identification of genetic elements associated with muscle structure in the nematode Caenorhabditis elegans. Cell Motil. 1980;1(1):73–97. doi: 10.1002/cm.970010107. [DOI] [PubMed] [Google Scholar]

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