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. 1985 Oct 1;101(4):1532–1549. doi: 10.1083/jcb.101.4.1532

Muscle organization in Caenorhabditis elegans: localization of proteins implicated in thin filament attachment and I-band organization

PMCID: PMC2113919  PMID: 2413045

Abstract

The body wall muscle cells of Caenorhabditis elegans contain an obliquely striated myofibrillar lattice that is associated with the cell membrane through two structures: an M-line analogue in the A-band and a Z-disc analogue, or dense-body, in the I-band. By using a fraction enriched in these structures as an immunogen for hybridoma production, we prepared monoclonal antibodies that identify four components of the I-band as determined by immunofluorescence and Western transfer analysis. A major constituent of the dense-body is a 107,000-D polypeptide that shares determinants with vertebrate alpha- actinin. A second dense-body constituent is a more basic and antigenically distinct 107,000-D polypeptide that is localized to a narrow domain of the dense-body at or subjacent to the plasma membrane. This basic dense-body polypeptide is also found at certain cell boundaries where thin filaments in half-bands terminate at membrane- associated structures termed attachment plaques. A third, unidentified antigen is also found closely apposed to the cell membrane in regions of not only the dense-body and attachment plaque, but also the M-line analogue. Finally, a fourth high molecular weight antigen, composed of two polypeptides of approximately 400,000-D, is localized to the I-band regions surrounding the dense-body. The attachment of the dense-body to the cell surface and the differential localization of the dense-body- associated antigens suggest a model for their organization in which the unidentified antigen is a cell surface component, and the two 107,000-D polypeptides define different cytoplasmic domains of the dense-body.

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Selected References

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  1. Albertson D. G. Localization of the ribosomal genes in Caenorhabditis elegans chromosomes by in situ hybridization using biotin-labeled probes. EMBO J. 1984 Jun;3(6):1227–1234. doi: 10.1002/j.1460-2075.1984.tb01957.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  2. Albertson D. G., Thomson J. N. The pharynx of Caenorhabditis elegans. Philos Trans R Soc Lond B Biol Sci. 1976 Aug 10;275(938):299–325. doi: 10.1098/rstb.1976.0085. [DOI] [PubMed] [Google Scholar]
  3. Batteiger B., Newhall W. J., 5th, Jones R. B. The use of Tween 20 as a blocking agent in the immunological detection of proteins transferred to nitrocellulose membranes. J Immunol Methods. 1982 Dec 30;55(3):297–307. doi: 10.1016/0022-1759(82)90089-8. [DOI] [PubMed] [Google Scholar]
  4. Blose S. H., Meltzer D. I., Feramisco J. R. 10-nm filaments are induced to collapse in living cells microinjected with monoclonal and polyclonal antibodies against tubulin. J Cell Biol. 1984 Mar;98(3):847–858. doi: 10.1083/jcb.98.3.847. [DOI] [PMC free article] [PubMed] [Google Scholar]
  5. Breckler J., Lazarides E. Isolation of a new high molecular weight protein associated with desmin and vimentin filaments from avian embryonic skeletal muscle. J Cell Biol. 1982 Mar;92(3):795–806. doi: 10.1083/jcb.92.3.795. [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. Bretscher A., Weber K. Localization of actin and microfilament-associated proteins in the microvilli and terminal web of the intestinal brush border by immunofluorescence microscopy. J Cell Biol. 1978 Dec;79(3):839–845. doi: 10.1083/jcb.79.3.839. [DOI] [PMC free article] [PubMed] [Google Scholar]
  8. Burridge K., Feramisco J. R. Microinjection and localization of a 130K protein in living fibroblasts: a relationship to actin and fibronectin. Cell. 1980 Mar;19(3):587–595. doi: 10.1016/s0092-8674(80)80035-3. [DOI] [PubMed] [Google Scholar]
  9. Chen W. T., Singer S. J. Immunoelectron microscopic studies of the sites of cell-substratum and cell-cell contacts in cultured fibroblasts. J Cell Biol. 1982 Oct;95(1):205–222. doi: 10.1083/jcb.95.1.205. [DOI] [PMC free article] [PubMed] [Google Scholar]
  10. Cox G. N., Kusch M., Edgar R. S. Cuticle of Caenorhabditis elegans: its isolation and partial characterization. J Cell Biol. 1981 Jul;90(1):7–17. doi: 10.1083/jcb.90.1.7. [DOI] [PMC free article] [PubMed] [Google Scholar]
  11. Craig R., Offer G. The location of C-protein in rabbit skeletal muscle. Proc R Soc Lond B Biol Sci. 1976 Mar 16;192(1109):451–461. doi: 10.1098/rspb.1976.0023. [DOI] [PubMed] [Google Scholar]
  12. Epstein H. F., Thomson J. N. Temperature-sensitive mutation affecting myofilament assembly in Caenorhabditis elegans. Nature. 1974 Aug 16;250(467):579–580. doi: 10.1038/250579a0. [DOI] [PubMed] [Google Scholar]
  13. Feramisco J. R., Burridge K. A rapid purification of alpha-actinin, filamin, and a 130,000-dalton protein from smooth muscle. J Biol Chem. 1980 Feb 10;255(3):1194–1199. [PubMed] [Google Scholar]
  14. Files J. G., Carr S., Hirsh D. Actin gene family of Caenorhabditis elegans. J Mol Biol. 1983 Mar 5;164(3):355–375. doi: 10.1016/0022-2836(83)90056-6. [DOI] [PubMed] [Google Scholar]
  15. Galfre G., Howe S. C., Milstein C., Butcher G. W., Howard J. C. Antibodies to major histocompatibility antigens produced by hybrid cell lines. Nature. 1977 Apr 7;266(5602):550–552. doi: 10.1038/266550a0. [DOI] [PubMed] [Google Scholar]
  16. Galfrè G., Milstein C. Preparation of monoclonal antibodies: strategies and procedures. Methods Enzymol. 1981;73(Pt B):3–46. doi: 10.1016/0076-6879(81)73054-4. [DOI] [PubMed] [Google Scholar]
  17. 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]
  18. Geiger B., Dutton A. H., Tokuyasu K. T., Singer S. J. Immunoelectron microscope studies of membrane-microfilament interactions: distributions of alpha-actinin, tropomyosin, and vinculin in intestinal epithelial brush border and chicken gizzard smooth muscle cells. J Cell Biol. 1981 Dec;91(3 Pt 1):614–628. doi: 10.1083/jcb.91.3.614. [DOI] [PMC free article] [PubMed] [Google Scholar]
  19. Gomer R. H., Lazarides E. Highly homologous filamin polypeptides have different distributions in avian slow and fast muscle fibers. J Cell Biol. 1983 Sep;97(3):818–823. doi: 10.1083/jcb.97.3.818. [DOI] [PMC free article] [PubMed] [Google Scholar]
  20. Gossett L. A., Hecht R. M., Epstein H. F. Muscle differentiation in normal and cleavage-arrested mutant embryos of Caenorhabditis elegans. Cell. 1982 Aug;30(1):193–204. doi: 10.1016/0092-8674(82)90025-3. [DOI] [PubMed] [Google Scholar]
  21. Granger B. L., Lazarides E. Synemin: a new high molecular weight protein associated with desmin and vimentin filaments in muscle. Cell. 1980 Dec;22(3):727–738. doi: 10.1016/0092-8674(80)90549-8. [DOI] [PubMed] [Google Scholar]
  22. Granger B. L., Lazarides E. The existence of an insoluble Z disc scaffold in chicken skeletal muscle. Cell. 1978 Dec;15(4):1253–1268. doi: 10.1016/0092-8674(78)90051-x. [DOI] [PubMed] [Google Scholar]
  23. 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]
  24. Harris H. E., Tso M. Y., Epstein H. F. Actin and myosin-linked calcium regulation in the nematode Caenorhabditis elegans. Biochemical and structural properties of native filaments and purified proteins. Biochemistry. 1977 Mar 8;16(5):859–865. doi: 10.1021/bi00624a008. [DOI] [PubMed] [Google Scholar]
  25. Hirumi H., Raski D. J., Jones N. O. Primitive muscle cells of nematodes: morphological aspects of platymyarian and shallow coelomyarian muscles in two plant parasitic nematodes, Trichodorus christiei and Longidorus elongatus. J Ultrastruct Res. 1971 Mar;34(5):517–543. doi: 10.1016/s0022-5320(71)80062-x. [DOI] [PubMed] [Google Scholar]
  26. Knight P. J., Trinick J. A. Preparation of myofibrils. Methods Enzymol. 1982;85(Pt B):9–12. doi: 10.1016/0076-6879(82)85004-0. [DOI] [PubMed] [Google Scholar]
  27. Krohne G., Stick R., Kleinschmidt J. A., Moll R., Franke W. W., Hausen P. Immunological localization of a major karyoskeletal protein in nucleoli of oocytes and somatic cells of Xenopus laevis. J Cell Biol. 1982 Sep;94(3):749–754. doi: 10.1083/jcb.94.3.749. [DOI] [PMC free article] [PubMed] [Google Scholar]
  28. Laemmli U. K. Cleavage of structural proteins during the assembly of the head of bacteriophage T4. Nature. 1970 Aug 15;227(5259):680–685. doi: 10.1038/227680a0. [DOI] [PubMed] [Google Scholar]
  29. Landel C. P., Krause M., Waterston R. H., Hirsh D. DNA rearrangements of the actin gene cluster in Caenorhabditis elegans accompany reversion of three muscle mutants. J Mol Biol. 1984 Dec 15;180(3):497–513. doi: 10.1016/0022-2836(84)90024-x. [DOI] [PubMed] [Google Scholar]
  30. Lazarides E., Burridge K. Alpha-actinin: immunofluorescent localization of a muscle structural protein in nonmuscle cells. Cell. 1975 Nov;6(3):289–298. doi: 10.1016/0092-8674(75)90180-4. [DOI] [PubMed] [Google Scholar]
  31. Lazarides E. Intermediate filaments: a chemically heterogeneous, developmentally regulated class of proteins. Annu Rev Biochem. 1982;51:219–250. doi: 10.1146/annurev.bi.51.070182.001251. [DOI] [PubMed] [Google Scholar]
  32. 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]
  33. 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]
  34. Mackenzie J. M., Jr, Garcea R. L., Zengel J. M., Epstein H. F. Muscle development in Caenorhabditis elegans: mutants exhibiting retarded sarcomere construction. Cell. 1978 Nov;15(3):751–762. doi: 10.1016/0092-8674(78)90261-1. [DOI] [PubMed] [Google Scholar]
  35. Mackenzie J. M., Jr, Schachat F., Epstein H. F. Immunocytochemical localization of two myosins within the same muslce cells in Caenorhabditis elegans. Cell. 1978 Oct;15(2):413–419. doi: 10.1016/0092-8674(78)90010-7. [DOI] [PubMed] [Google Scholar]
  36. Masaki T., Endo M., Ebashi S. Localization of 6S component of a alpha-actinin at Z-band. J Biochem. 1967 Nov;62(5):630–632. doi: 10.1093/oxfordjournals.jbchem.a128717. [DOI] [PubMed] [Google Scholar]
  37. 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]
  38. 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]
  39. Moerman D. G., Baillie D. L. Formaldehyde mutagenesis in the nematode Caenorhabditis elegans. Mutat Res. 1981 Feb;80(2):273–279. doi: 10.1016/0027-5107(81)90100-7. [DOI] [PubMed] [Google Scholar]
  40. 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]
  41. O'Farrell P. H. High resolution two-dimensional electrophoresis of proteins. J Biol Chem. 1975 May 25;250(10):4007–4021. [PMC free article] [PubMed] [Google Scholar]
  42. Rosenbluth J. Obliquely striated muscle. 3. Contraction mechanism of Ascaris body muscle. J Cell Biol. 1967 Jul;34(1):15–33. doi: 10.1083/jcb.34.1.15. [DOI] [PMC free article] [PubMed] [Google Scholar]
  43. Rosenbluth J. Ultrastructural organization of obliquely striated muscle fibers in Ascaris lumbricoides. J Cell Biol. 1965 Jun;25(3):495–515. doi: 10.1083/jcb.25.3.495. [DOI] [PMC free article] [PubMed] [Google Scholar]
  44. Shulman M., Wilde C. D., Köhler G. A better cell line for making hybridomas secreting specific antibodies. Nature. 1978 Nov 16;276(5685):269–270. doi: 10.1038/276269a0. [DOI] [PubMed] [Google Scholar]
  45. Sulston J. E., Brenner S. The DNA of Caenorhabditis elegans. Genetics. 1974 May;77(1):95–104. doi: 10.1093/genetics/77.1.95. [DOI] [PMC free article] [PubMed] [Google Scholar]
  46. Sulston J. E., Horvitz H. R. Post-embryonic cell lineages of the nematode, Caenorhabditis elegans. Dev Biol. 1977 Mar;56(1):110–156. doi: 10.1016/0012-1606(77)90158-0. [DOI] [PubMed] [Google Scholar]
  47. 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]
  48. Suzuki A., Goll D. E., Singh I., Allen R. E., Robson R. M., Stromer M. H. Some properties of purified skeletal muscle alpha-actinin. J Biol Chem. 1976 Nov 10;251(21):6860–6870. [PubMed] [Google Scholar]
  49. Tokuyasu K. T., Dutton A. H., Geiger B., Singer S. J. Ultrastructure of chicken cardiac muscle as studied by double immunolabeling in electron microscopy. Proc Natl Acad Sci U S A. 1981 Dec;78(12):7619–7623. doi: 10.1073/pnas.78.12.7619. [DOI] [PMC free article] [PubMed] [Google Scholar]
  50. Towbin H., Staehelin T., Gordon J. Electrophoretic transfer of proteins from polyacrylamide gels to nitrocellulose sheets: procedure and some applications. Proc Natl Acad Sci U S A. 1979 Sep;76(9):4350–4354. doi: 10.1073/pnas.76.9.4350. [DOI] [PMC free article] [PubMed] [Google Scholar]
  51. Trinick J., Lowey S. M-protein from chicken pectoralis muscle: isolation and characterization. J Mol Biol. 1977 Jun 25;113(2):343–368. doi: 10.1016/0022-2836(77)90146-2. [DOI] [PubMed] [Google Scholar]
  52. Wallimann T., Turner D. C., Eppenberger H. M. Localization of creatine kinase isoenzymes in myofibrils. I. Chicken skeletal muscle. J Cell Biol. 1977 Nov;75(2 Pt 1):297–317. doi: 10.1083/jcb.75.2.297. [DOI] [PMC free article] [PubMed] [Google Scholar]
  53. Wang K. Cytoskeletal matrix in striated muscle: the role of titin, nebulin and intermediate filaments. Adv Exp Med Biol. 1984;170:285–305. doi: 10.1007/978-1-4684-4703-3_25. [DOI] [PubMed] [Google Scholar]
  54. Wang K., McClure J., Tu A. Titin: major myofibrillar components of striated muscle. Proc Natl Acad Sci U S A. 1979 Aug;76(8):3698–3702. doi: 10.1073/pnas.76.8.3698. [DOI] [PMC free article] [PubMed] [Google Scholar]
  55. Wang K., Williamson C. L. Identification of an N2 line protein of striated muscle. Proc Natl Acad Sci U S A. 1980 Jun;77(6):3254–3258. doi: 10.1073/pnas.77.6.3254. [DOI] [PMC free article] [PubMed] [Google Scholar]
  56. 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]
  57. Waterston R. H., Hirsh D., Lane T. R. Dominant mutations affecting muscle structure in Caenorhabditis elegans that map near the actin gene cluster. J Mol Biol. 1984 Dec 15;180(3):473–496. doi: 10.1016/0022-2836(84)90023-8. [DOI] [PubMed] [Google Scholar]
  58. 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]
  59. Wehland J., Osborn M., Weber K. Cell-to-substratum contacts in living cells: a direct correlation between interference-reflexion and indirect-immunofluorescence microscopy using antibodies against actin and alpha-actinin. J Cell Sci. 1979 Jun;37:257–273. doi: 10.1242/jcs.37.1.257. [DOI] [PubMed] [Google Scholar]
  60. White J. G., Southgate E., Thomson J. N., Brenner S. The structure of the ventral nerve cord of Caenorhabditis elegans. Philos Trans R Soc Lond B Biol Sci. 1976 Aug 10;275(938):327–348. doi: 10.1098/rstb.1976.0086. [DOI] [PubMed] [Google Scholar]
  61. Wiche G., Herrmann H., Leichtfried F., Pytela R. Plectin: a high-molecular-weight cytoskeletal polypeptide component that copurifies with intermediate filaments of the vimentin type. Cold Spring Harb Symp Quant Biol. 1982;46(Pt 1):475–482. doi: 10.1101/sqb.1982.046.01.044. [DOI] [PubMed] [Google Scholar]
  62. Wulf E., Deboben A., Bautz F. A., Faulstich H., Wieland T. Fluorescent phallotoxin, a tool for the visualization of cellular actin. Proc Natl Acad Sci U S A. 1979 Sep;76(9):4498–4502. doi: 10.1073/pnas.76.9.4498. [DOI] [PMC free article] [PubMed] [Google Scholar]
  63. Young R. A., Davis R. W. Efficient isolation of genes by using antibody probes. Proc Natl Acad Sci U S A. 1983 Mar;80(5):1194–1198. doi: 10.1073/pnas.80.5.1194. [DOI] [PMC free article] [PubMed] [Google Scholar]
  64. 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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