Skip to main content
The EMBO Journal logoLink to The EMBO Journal
. 1989 Nov;8(11):3429–3436. doi: 10.1002/j.1460-2075.1989.tb08507.x

The minor myosin heavy chain, mhcA, of Caenorhabditis elegans is necessary for the initiation of thick filament assembly.

R H Waterston 1
PMCID: PMC401495  PMID: 2583106

Abstract

Caenorhabditis elegans body wall muscle has two distinct myosin heavy chain isoforms, mhcA and mhcB. Mutations eliminating the major isoform, mhcB, have previously been shown to yield paralyzed, viable animals. In this paper we show that the minor isoform, mhcA, is essential for viability. We have utilized the known physical map position of the gene encoding mhcA to obtain two recessive lethal mutations that virtually eliminate accumulation of mhcA. The mutations are allelic, and the interactions of these alleles with mutations affecting other thick filament components are consistent with the hypothesis that the new mutations lie in the structural gene for mhcA. The homozygous mutant animals move very little and morphological analysis shows that thick filament assembly is severely impaired. Together with the location of mhcA in the center of the thick filament (Miller et al., 1983), the results suggest that mhcA has a unique role in initiating filament assembly. The homozygous mutations have an unexpected effect on morphogenesis that indicates an interaction between the muscle cells and the hypodermis during development. The resultant phenotype may be useful in the search for additional essential muscle genes.

Full text

PDF
3429

Images in this article

Selected References

These references are in PubMed. This may not be the complete list of references from this article.

  1. Albertson D. G. Mapping muscle protein genes by in situ hybridization using biotin-labeled probes. EMBO J. 1985 Oct;4(10):2493–2498. doi: 10.1002/j.1460-2075.1985.tb03961.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  2. Anderson P., Brenner S. A selection for myosin heavy chain mutants in the nematode Caenorhabditis elegans. Proc Natl Acad Sci U S A. 1984 Jul;81(14):4470–4474. doi: 10.1073/pnas.81.14.4470. [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. Bejsovec A., Anderson P. Myosin heavy-chain mutations that disrupt Caenorhabditis elegans thick filament assembly. Genes Dev. 1988 Oct;2(10):1307–1317. doi: 10.1101/gad.2.10.1307. [DOI] [PubMed] [Google Scholar]
  5. Bernstein S. I., Hansen C. J., Becker K. D., Wassenberg D. R., 2nd, Roche E. S., Donady J. J., Emerson C. P., Jr Alternative RNA splicing generates transcripts encoding a thorax-specific isoform of Drosophila melanogaster myosin heavy chain. Mol Cell Biol. 1986 Jul;6(7):2511–2519. doi: 10.1128/mcb.6.7.2511. [DOI] [PMC free article] [PubMed] [Google Scholar]
  6. 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]
  7. 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]
  8. 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]
  9. Dibb N. J., Brown D. M., Karn J., Moerman D. G., Bolten S. L., Waterston R. H. Sequence analysis of mutations that affect the synthesis, assembly and enzymatic activity of the unc-54 myosin heavy chain of Caenorhabditis elegans. J Mol Biol. 1985 Jun 25;183(4):543–551. doi: 10.1016/0022-2836(85)90170-6. [DOI] [PubMed] [Google Scholar]
  10. Epstein H. F., Miller D. M., 3rd, Ortiz I., Berliner G. C. Myosin and paramyosin are organized about a newly identified core structure. J Cell Biol. 1985 Mar;100(3):904–915. doi: 10.1083/jcb.100.3.904. [DOI] [PMC free article] [PubMed] [Google Scholar]
  11. Epstein H. F., Waterston R. H., Brenner S. A mutant affecting the heavy chain of myosin in Caenorhabditis elegans. J Mol Biol. 1974 Dec 5;90(2):291–300. doi: 10.1016/0022-2836(74)90374-x. [DOI] [PubMed] [Google Scholar]
  12. Ferguson E. L., Horvitz H. R. Identification and characterization of 22 genes that affect the vulval cell lineages of the nematode Caenorhabditis elegans. Genetics. 1985 May;110(1):17–72. doi: 10.1093/genetics/110.1.17. [DOI] [PMC free article] [PubMed] [Google Scholar]
  13. Fire A., Waterston R. H. Proper expression of myosin genes in transgenic nematodes. EMBO J. 1989 Nov;8(11):3419–3428. doi: 10.1002/j.1460-2075.1989.tb08506.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  14. 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]
  15. Garoff H., Ansorge W. Improvements of DNA sequencing gels. Anal Biochem. 1981 Aug;115(2):450–457. doi: 10.1016/0003-2697(81)90031-2. [DOI] [PubMed] [Google Scholar]
  16. Kagawa H., Gengyo K., McLachlan A. D., Brenner S., Karn J. Paramyosin gene (unc-15) of Caenorhabditis elegans. Molecular cloning, nucleotide sequence and models for thick filament structure. J Mol Biol. 1989 May 20;207(2):311–333. doi: 10.1016/0022-2836(89)90257-x. [DOI] [PubMed] [Google Scholar]
  17. MacLeod A. R., Waterston R. H., Fishpool R. M., Brenner S. Identification of the structural gene for a myosin heavy-chain in Caenorhabditis elegans. J Mol Biol. 1977 Jul;114(1):133–140. doi: 10.1016/0022-2836(77)90287-x. [DOI] [PubMed] [Google Scholar]
  18. Mackenzie J. M., Jr, Epstein H. F. Paramyosin is necessary for determination of nematode thick filament length in vivo. Cell. 1980 Dec;22(3):747–755. doi: 10.1016/0092-8674(80)90551-6. [DOI] [PubMed] [Google Scholar]
  19. McGhee J. D., Cottrell D. A. The major gut esterase locus in the nematode Caenorhabditis elegans. Mol Gen Genet. 1986 Jan;202(1):30–34. doi: 10.1007/BF00330512. [DOI] [PubMed] [Google Scholar]
  20. Merril C. R., Goldman D., Sedman S. A., Ebert M. H. Ultrasensitive stain for proteins in polyacrylamide gels shows regional variation in cerebrospinal fluid proteins. Science. 1981 Mar 27;211(4489):1437–1438. doi: 10.1126/science.6162199. [DOI] [PubMed] [Google Scholar]
  21. 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]
  22. Miller D. M., Stockdale F. E., Karn J. Immunological identification of the genes encoding the four myosin heavy chain isoforms of Caenorhabditis elegans. Proc Natl Acad Sci U S A. 1986 Apr;83(8):2305–2309. doi: 10.1073/pnas.83.8.2305. [DOI] [PMC free article] [PubMed] [Google Scholar]
  23. Moerman D. G., Plurad S., Waterston R. H., Baillie D. L. Mutations in the unc-54 myosin heavy chain gene of Caenorhabditis elegans that alter contractility but not muscle structure. Cell. 1982 Jul;29(3):773–781. doi: 10.1016/0092-8674(82)90439-1. [DOI] [PubMed] [Google Scholar]
  24. 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]
  25. Riddle D. L., Brenner S. Indirect suppression in Caenorhabditis elegans. Genetics. 1978 Jun;89(2):299–314. doi: 10.1093/genetics/89.2.299. [DOI] [PMC free article] [PubMed] [Google Scholar]
  26. Rozek C. E., Davidson N. Drosophila has one myosin heavy-chain gene with three developmentally regulated transcripts. Cell. 1983 Jan;32(1):23–34. doi: 10.1016/0092-8674(83)90493-2. [DOI] [PubMed] [Google Scholar]
  27. Schachat F., Garcea R. L., Epstein H. F. Myosins exist as homodimers of heavy chains: demonstration with specific antibody purified by nematode mutant myosin affinity chromatography. Cell. 1978 Oct;15(2):405–411. doi: 10.1016/0092-8674(78)90009-0. [DOI] [PubMed] [Google Scholar]
  28. 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]
  29. Sulston J., Dew M., Brenner S. Dopaminergic neurons in the nematode Caenorhabditis elegans. J Comp Neurol. 1975 Sep 15;163(2):215–226. doi: 10.1002/cne.901630207. [DOI] [PubMed] [Google Scholar]
  30. Warrick H. M., Spudich J. A. Myosin structure and function in cell motility. Annu Rev Cell Biol. 1987;3:379–421. doi: 10.1146/annurev.cb.03.110187.002115. [DOI] [PubMed] [Google Scholar]
  31. 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]
  32. Waterston R. H., Fishpool R. M., Brenner S. Mutants affecting paramyosin in Caenorhabditis elegans. J Mol Biol. 1977 Dec 15;117(3):679–697. doi: 10.1016/0022-2836(77)90064-x. [DOI] [PubMed] [Google Scholar]
  33. Waterston R. H., Thomson J. N., Brenner S. Mutants with altered muscle structure of Caenorhabditis elegans. Dev Biol. 1980 Jun 15;77(2):271–302. doi: 10.1016/0012-1606(80)90475-3. [DOI] [PubMed] [Google Scholar]

Articles from The EMBO Journal are provided here courtesy of Nature Publishing Group

RESOURCES