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. 1993 Oct 2;123(2):303–311. doi: 10.1083/jcb.123.2.303

Thick filament substructures in Caenorhabditis elegans: evidence for two populations of paramyosin

PMCID: PMC2119837  PMID: 8408214

Abstract

The thick filaments of the nematode Caenorhabditis elegans contain two myosin heavy chain isoforms A and B and paramyosin, the products of the myo-3, unc-54, and unc-15 genes, respectively. Dissociation of paramyosin from native thick filaments at pH 6.36 shows a biphasic function with respect to NaCl concentration. Electron microscopy of the remaining structures shows 15-nm core structures that label with monoclonal anti-paramyosin antibody at 72.5-nm intervals. Purified core structures also show 72.5 nm repeats by negative staining. Structural analysis of native thick filaments and dissociated structures suggests that the more dissociable paramyosin is removed radially as well as processively from the filament ends. Minor proteins with masses of 20, 28, and 30 kD cosediment stoichiometrically with paramyosin in purified core structures.

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Selected References

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  1. Anderson P. Molecular genetics of nematode muscle. Annu Rev Genet. 1989;23:507–525. doi: 10.1146/annurev.ge.23.120189.002451. [DOI] [PubMed] [Google Scholar]
  2. Becker K. D., O'Donnell P. T., Heitz J. M., Vito M., Bernstein S. I. Analysis of Drosophila paramyosin: identification of a novel isoform which is restricted to a subset of adult muscles. J Cell Biol. 1992 Feb;116(3):669–681. doi: 10.1083/jcb.116.3.669. [DOI] [PMC free article] [PubMed] [Google Scholar]
  3. Castellani L., Vibert P. Location of paramyosin in relation to the subfilaments within the thick filaments of scallop striated muscle. J Muscle Res Cell Motil. 1992 Apr;13(2):174–182. doi: 10.1007/BF01874154. [DOI] [PubMed] [Google Scholar]
  4. Cohen C., Szent-Györgyi A. G., Kendrick-Jones J. Paramyosin and the filaments of molluscan "catch" muscles. I. Paramyosin: structure and assembly. J Mol Biol. 1971 Mar 14;56(2):223–227. doi: 10.1016/0022-2836(71)90461-x. [DOI] [PubMed] [Google Scholar]
  5. Davis J. S. A model for length-regulation in thick filaments of vertebrate skeletal myosin. Biophys J. 1986 Sep;50(3):417–422. doi: 10.1016/S0006-3495(86)83477-4. [DOI] [PMC free article] [PubMed] [Google Scholar]
  6. Dey C. S., Deitiker P. R., Epstein H. F. Assembly-dependent phosphorylation of myosin and paramyosin of native thick filaments in Caenorhabditis elegans. Biochem Biophys Res Commun. 1992 Aug 14;186(3):1528–1532. doi: 10.1016/s0006-291x(05)81580-3. [DOI] [PubMed] [Google Scholar]
  7. Epstein H. F., Berliner G. C., Casey D. L., Ortiz I. Purified thick filaments from the nematode Caenorhabditis elegans: evidence for multiple proteins associated with core structures. J Cell Biol. 1988 Jun;106(6):1985–1995. doi: 10.1083/jcb.106.6.1985. [DOI] [PMC free article] [PubMed] [Google Scholar]
  8. Epstein H. F., Casey D. L., Ortiz I. Myosin and paramyosin of Caenorhabditis elegans embryos assemble into nascent structures distinct from thick filaments and multi-filament assemblages. J Cell Biol. 1993 Aug;122(4):845–858. doi: 10.1083/jcb.122.4.845. [DOI] [PMC free article] [PubMed] [Google Scholar]
  9. Epstein H. F., Fischman D. A. Molecular analysis of protein assembly in muscle development. Science. 1991 Mar 1;251(4997):1039–1044. doi: 10.1126/science.1998120. [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., Ortiz I., Berliner G. C. Assemblages of multiple thick filaments in nematode mutants. J Muscle Res Cell Motil. 1987 Dec;8(6):527–536. doi: 10.1007/BF01567911. [DOI] [PubMed] [Google Scholar]
  12. Epstein H. F., Ortiz I., Mackinnon L. A. The alteration of myosin isoform compartmentation in specific mutants of Caenorhabditis elegans. J Cell Biol. 1986 Sep;103(3):985–993. doi: 10.1083/jcb.103.3.985. [DOI] [PMC free article] [PubMed] [Google Scholar]
  13. Gengyo-Ando K., Kagawa H. Single charge change on the helical surface of the paramyosin rod dramatically disrupts thick filament assembly in Caenorhabditis elegans. J Mol Biol. 1991 Jun 5;219(3):429–441. doi: 10.1016/0022-2836(91)90184-8. [DOI] [PubMed] [Google Scholar]
  14. Harris H. E., Epstein H. F. Myosin and paramyosin of Caenorhabditis elegans: biochemical and structural properties of wild-type and mutant proteins. Cell. 1977 Apr;10(4):709–719. doi: 10.1016/0092-8674(77)90105-2. [DOI] [PubMed] [Google Scholar]
  15. Honda S., Epstein H. F. Modulation of muscle gene expression in Caenorhabditis elegans: differential levels of transcripts, mRNAs, and polypeptides for thick filament proteins during nematode development. Proc Natl Acad Sci U S A. 1990 Feb;87(3):876–880. doi: 10.1073/pnas.87.3.876. [DOI] [PMC free article] [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. Levine R. J., Elfvin M., Dewey M. M., Walcott B. Paramyosin in invertebrate muscles. II. Content in relation to structure and function. J Cell Biol. 1976 Oct;71(1):273–279. doi: 10.1083/jcb.71.1.273. [DOI] [PMC free article] [PubMed] [Google Scholar]
  18. Levine R. J., Kensler R. W., Levitt P. Crossbridge and backbone structure of invertebrate thick filaments. Biophys J. 1986 Jan;49(1):135–138. doi: 10.1016/S0006-3495(86)83624-4. [DOI] [PMC free article] [PubMed] [Google Scholar]
  19. Levine R. J., Kensler R. W., Reedy M. C., Hofmann W., King H. A. Structure and paramyosin content of tarantula thick filaments. J Cell Biol. 1983 Jul;97(1):186–195. doi: 10.1083/jcb.97.1.186. [DOI] [PMC free article] [PubMed] [Google Scholar]
  20. Levine R. J., Kensler R. W., Stewart M., Haselgrove J. C. Molecular organization of Limulus thick filaments. Soc Gen Physiol Ser. 1982;37:37–52. [PubMed] [Google Scholar]
  21. 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]
  22. McLachlan A. D., Karn J. Periodic charge distributions in the myosin rod amino acid sequence match cross-bridge spacings in muscle. Nature. 1982 Sep 16;299(5880):226–231. doi: 10.1038/299226a0. [DOI] [PubMed] [Google Scholar]
  23. McLachlan A. D., Karn J. Periodic features in the amino acid sequence of nematode myosin rod. J Mol Biol. 1983 Mar 15;164(4):605–626. doi: 10.1016/0022-2836(83)90053-0. [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. Morimoto K., Harrington W. F. Isolation and composition of thick filaments from rabbit skeletal muscle. J Mol Biol. 1973 Jun 15;77(1):165–175. doi: 10.1016/0022-2836(73)90370-7. [DOI] [PubMed] [Google Scholar]
  26. 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]
  27. Schriefer L. A., Waterson R. H. Phosphorylation of the N-terminal region of Caenorhabditis elegans paramyosin. J Mol Biol. 1989 May 20;207(2):451–454. doi: 10.1016/0022-2836(89)90267-2. [DOI] [PubMed] [Google Scholar]
  28. Szent-Györgyi A. G., Cohen C., Kendrick-Jones J. Paramyosin and the filaments of molluscan "catch" muscles. II. Native filaments: isolation and characterization. J Mol Biol. 1971 Mar 14;56(2):239–258. doi: 10.1016/0022-2836(71)90462-1. [DOI] [PubMed] [Google Scholar]
  29. Vibert P., Castellani L. Substructure and accessory proteins in scallop myosin filaments. J Cell Biol. 1989 Aug;109(2):539–547. doi: 10.1083/jcb.109.2.539. [DOI] [PMC free article] [PubMed] [Google Scholar]
  30. Waterston R. H., Epstein H. F., Brenner S. Paramyosin of Caenorhabditis elegans. J Mol Biol. 1974 Dec 5;90(2):285–290. doi: 10.1016/0022-2836(74)90373-8. [DOI] [PubMed] [Google Scholar]
  31. 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]
  32. 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]

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