Abstract
The precise arrangement of molecules within the thick filament, as well as the mechanisms by which this arrangement is specified, remains unclear. In this article, we have exploited a unique genetic interaction between one isoform of myosin heavy chain (MHC) and paramyosin in Caenorhabditis elegans to probe the molecular interaction between MHC and paramyosin in vivo. Using chimeric myosin constructs, we have defined a 322-residue region of the MHC A rod critical for suppression of the structural and motility defects associated with the unc-15(e73) allele. Chimeric constructs lacking this region of MHC A either fail to suppress, or act as dominant enhancers of, the e73 phenotype. Although the 322-residue region is required for suppression activity, our data suggest that sequences along the length of the rod also play a role in the isoform-specific interaction between MHC A and paramyosin. Our genetic and cell biological analyses of construct behavior suggest that the 322-residue region of MHC A is important for thick filament stability. We present a model in which this region mediates an avid interaction between MHC A and paramyosin in parallel arrangement in formation of the filament arms.
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- Ao W., Pilgrim D. Caenorhabditis elegans UNC-45 is a component of muscle thick filaments and colocalizes with myosin heavy chain B, but not myosin heavy chain A. J Cell Biol. 2000 Jan 24;148(2):375–384. doi: 10.1083/jcb.148.2.375. [DOI] [PMC free article] [PubMed] [Google Scholar]
- 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]
- Barral J. M., Bauer C. C., Ortiz I., Epstein H. F. Unc-45 mutations in Caenorhabditis elegans implicate a CRO1/She4p-like domain in myosin assembly. J Cell Biol. 1998 Nov 30;143(5):1215–1225. doi: 10.1083/jcb.143.5.1215. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Barral J. M., Epstein H. F. Protein machines and self assembly in muscle organization. Bioessays. 1999 Oct;21(10):813–823. doi: 10.1002/(SICI)1521-1878(199910)21:10<813::AID-BIES3>3.0.CO;2-0. [DOI] [PubMed] [Google Scholar]
- Barstead R. J., Waterston R. H. Vinculin is essential for muscle function in the nematode. J Cell Biol. 1991 Aug;114(4):715–724. doi: 10.1083/jcb.114.4.715. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Brown S. J., Riddle D. L. Gene interactions affecting muscle organization in Caenorhabditis elegans. Genetics. 1985 Jul;110(3):421–440. doi: 10.1093/genetics/110.3.421. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Cohen C., Lanar D. E., Parry D. A. Amino acid sequence and structural repeats in schistosome paramyosin match those of myosin. Biosci Rep. 1987 Jan;7(1):11–16. doi: 10.1007/BF01122722. [DOI] [PubMed] [Google Scholar]
- Cohen C., Parry D. A. A conserved C-terminal assembly region in paramyosin and myosin rods. J Struct Biol. 1998;122(1-2):180–187. doi: 10.1006/jsbi.1998.3983. [DOI] [PubMed] [Google Scholar]
- Deitiker P. R., Epstein H. F. Thick filament substructures in Caenorhabditis elegans: evidence for two populations of paramyosin. J Cell Biol. 1993 Oct;123(2):303–311. doi: 10.1083/jcb.123.2.303. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Epstein H. F., Lu G. Y., Deitiker P. R., Oritz I., Schmid M. F. Preliminary three-dimensional model for nematode thick filament core. J Struct Biol. 1995 Sep-Oct;115(2):163–174. doi: 10.1006/jsbi.1995.1041. [DOI] [PubMed] [Google Scholar]
- 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]
- 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]
- 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]
- 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]
- 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]
- 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]
- Francis R., Maine E., Schedl T. Analysis of the multiple roles of gld-1 in germline development: interactions with the sex determination cascade and the glp-1 signaling pathway. Genetics. 1995 Feb;139(2):607–630. doi: 10.1093/genetics/139.2.607. [DOI] [PMC free article] [PubMed] [Google Scholar]
- 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]
- 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]
- Hoppe P. E., Waterston R. H. Hydrophobicity variations along the surface of the coiled-coil rod may mediate striated muscle myosin assembly in Caenorhabditis elegans. J Cell Biol. 1996 Oct;135(2):371–382. doi: 10.1083/jcb.135.2.371. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Horton R. M., Hunt H. D., Ho S. N., Pullen J. K., Pease L. R. Engineering hybrid genes without the use of restriction enzymes: gene splicing by overlap extension. Gene. 1989 Apr 15;77(1):61–68. doi: 10.1016/0378-1119(89)90359-4. [DOI] [PubMed] [Google Scholar]
- Hresko M. C., Williams B. D., Waterston R. H. Assembly of body wall muscle and muscle cell attachment structures in Caenorhabditis elegans. J Cell Biol. 1994 Feb;124(4):491–506. doi: 10.1083/jcb.124.4.491. [DOI] [PMC free article] [PubMed] [Google Scholar]
- 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]
- Liu F., Barral J. M., Bauer C. C., Ortiz I., Cook R. G., Schmid M. F., Epstein H. F. Assemblases and coupling proteins in thick filament assembly. Cell Struct Funct. 1997 Feb;22(1):155–162. doi: 10.1247/csf.22.155. [DOI] [PubMed] [Google Scholar]
- Liu F., Bauer C. C., Ortiz I., Cook R. G., Schmid M. F., Epstein H. F. beta-Filagenin, a newly identified protein coassembling with myosin and paramyosin in Caenorhabditis elegans. J Cell Biol. 1998 Jan 26;140(2):347–353. doi: 10.1083/jcb.140.2.347. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Lowey S., Slayter H. S., Weeds A. G., Baker H. Substructure of the myosin molecule. I. Subfragments of myosin by enzymic degradation. J Mol Biol. 1969 May 28;42(1):1–29. doi: 10.1016/0022-2836(69)90483-5. [DOI] [PubMed] [Google Scholar]
- 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]
- Maruyama I. N., Miller D. M., Brenner S. Myosin heavy chain gene amplification as a suppressor mutation in Caenorhabditis elegans. Mol Gen Genet. 1989 Oct;219(1-2):113–118. doi: 10.1007/BF00261165. [DOI] [PubMed] [Google Scholar]
- 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]
- Mello C. C., Kramer J. M., Stinchcomb D., Ambros V. Efficient gene transfer in C.elegans: extrachromosomal maintenance and integration of transforming sequences. EMBO J. 1991 Dec;10(12):3959–3970. doi: 10.1002/j.1460-2075.1991.tb04966.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Park Y. S., Kramer J. M. The C. elegans sqt-1 and rol-6 collagen genes are coordinately expressed during development, but not at all stages that display mutant phenotypes. Dev Biol. 1994 May;163(1):112–124. doi: 10.1006/dbio.1994.1127. [DOI] [PubMed] [Google Scholar]
- 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]
- 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]
- Sinard J. H., Rimm D. L., Pollard T. D. Identification of functional regions on the tail of Acanthamoeba myosin-II using recombinant fusion proteins. II. Assembly properties of tails with NH2- and COOH-terminal deletions. J Cell Biol. 1990 Dec;111(6 Pt 1):2417–2426. doi: 10.1083/jcb.111.6.2417. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Sinard J. H., Stafford W. F., Pollard T. D. The mechanism of assembly of Acanthamoeba myosin-II minifilaments: minifilaments assemble by three successive dimerization steps. J Cell Biol. 1989 Oct;109(4 Pt 1):1537–1547. doi: 10.1083/jcb.109.4.1537. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Sohn R. L., Vikstrom K. L., Strauss M., Cohen C., Szent-Gyorgyi A. G., Leinwand L. A. A 29 residue region of the sarcomeric myosin rod is necessary for filament formation. J Mol Biol. 1997 Feb 21;266(2):317–330. doi: 10.1006/jmbi.1996.0790. [DOI] [PubMed] [Google Scholar]
- 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]
- Venolia L., Ao W., Kim S., Kim C., Pilgrim D. unc-45 gene of Caenorhabditis elegans encodes a muscle-specific tetratricopeptide repeat-containing protein. Cell Motil Cytoskeleton. 1999;42(3):163–177. doi: 10.1002/(SICI)1097-0169(1999)42:3<163::AID-CM1>3.0.CO;2-E. [DOI] [PubMed] [Google Scholar]
- Venolia L., Waterston R. H. The unc-45 gene of Caenorhabditis elegans is an essential muscle-affecting gene with maternal expression. Genetics. 1990 Oct;126(2):345–353. doi: 10.1093/genetics/126.2.345. [DOI] [PMC free article] [PubMed] [Google Scholar]
- 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]
- 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]
- Wells L., Edwards K. A., Bernstein S. I. Myosin heavy chain isoforms regulate muscle function but not myofibril assembly. EMBO J. 1996 Sep 2;15(17):4454–4459. [PMC free article] [PubMed] [Google Scholar]
- Williams B. D., Waterston R. H. Genes critical for muscle development and function in Caenorhabditis elegans identified through lethal mutations. J Cell Biol. 1994 Feb;124(4):475–490. doi: 10.1083/jcb.124.4.475. [DOI] [PMC free article] [PubMed] [Google Scholar]