Abstract
The established observations and unresolved questions in the assembly of myosin are outlined in this article. Much of the background information has been obtained in classical experiments using the myosin and thick filaments from vertebrate skeletal muscle. Current research is concerned with problems of myosin assembly and structure in smooth muscle, a broad spectrum of invertebrate muscles, and eukaryotic cells in general.
Many of the general questions concerning myosin assembly have been addressed by a combination of genetic, molecular, and structural approaches in the nematodeCaenorhabditis elegans. Detailed analysis of multiple myosin isoforms has been a prominent aspect of the nematode work. The molecular cloning and determination of the complete sequences of the genes encoding the four isoforms of myosin heavy chain and of the myosinassociated protein paramyosin have been a major landmark.
The sequences have permitted a theoretical analysis of myosin rod structure and the interactions of myosin in thick filaments. The development of specific monoclonal antibodies to the individual myosins has led to the delineation of the different locations of the myosins and to their special roles in thick filament structure and assembly.
In nematode body-wall muscles, two isoforms, myosins A and B, are located in different regions of each thick filament. Myosin A is located in the central biopolar zones, whereas myosin B is restricted to the flanking polar regions. This specific localization directly implies differential behavior of the two myosins during assembly. Genetic and structural experiments demonstrate that paramyosin and the levels of expression of the two forms are required for the differential assembly. Additional genetic experiments indicate that several other gene products are involved in the assembly of myosin. Structural studies of mutants have uncovered two new structures. A core structure separate from myosin and paramyosin appears to be an integral part of thick filaments. Multifilament assemblages exhibit multiple nascent thick filament-like structures extending from central paramyosin regions. Dominant mutants of myosin that disrupt thick filament assembly are located in the ATP and actin binding sites of the heavy chain.
A model for a cycle of reactions in the assembly of myosin into thick filaments is presented. Specific reactions of the two myosin isoforms, paramyosin, and core proteins with multifilament assemblages as possible intermediates in assembly are proposed.
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References
Albertson D. G. (1985) Mapping muscle protein genes byin situ hybridization using biotin-labeled probes.EMBO J. 4, 2493–2498.
Anderson, P. (1989) Molecular genetics of nematode muscle.Annu. Rev. Genet. 23, 507–525.
Anderson P. and Brenner S. (1984) A selection for myosin heavy-chain mutants in the nematode.Caenorhabditis elegans.Proc. Natl. Acad. Sci. USA 81, 4470–4474.
Ardizzi J. P. and Epstein H. F. (1987) Immunochemical localization of myosin heavy chain isoforms and paramyosin of developmentally and structurally diverse muscle cell types of the nematodeCaenorhabditis elegans.J. Cell Biol. 105, 2763–2770.
Bejsovec A. and Anderson P. (1988) Myosin heavy chain mutations that disruptCaenorhabditis elegans thick filament assembly.Genes Devel. 2, 1307–1317.
Bejsovec A. and Anderson P. (1990) Function of the myosin ATPase and actin binding sites is required forC. elegans thick filament assembly.Cell 60, 133–140.
Bejsovec A., Eide D., and Anderson P. (1989) Genetic techniques for analysis of nematode muscle, inMolecular Biology of the Cytoskeleton, Borisy G. et al., eds., Cold Spring Harbor Laboratory, Cold Spring Harbor, New York, pp. 267–273.
Benian G. M., Kiff J. E., Neckelmann N., Moerman D. G., and Waterston, R. H. (1989) Sequence of an unusually large protein implicated in regulation of myosin activity inC. elegans.Nature 342, 45–50.
Bennett P., Craig R., Starr R., and Offer G. (1986) The ultrastructural localization of C-protein, X-protein and H-protein in rabbit muscle.J. Muscle Res. Cell Motil. 7, 556–567.
Bouché M., Goldfine S. M., and Fischman D. A. (1988) Posttranslational incorporation of contractile proteins into myofibrils in a cell-free system.J. Cell Biol. 107, 587–596.
Brown S. J. and Riddle D. L. (1985) Gene interactions affecting muscle organization inCaenorhabditis elegans.Genetics 110, 421–440.
Bullard B., Luke B., and Winkelman L. (1973) The paramyosin of insect flight muscle.J. Mol. Biol. 75, 359–367.
Chowrashi P. K. and Pepe F. A. (1986) The myosin filament XII. Effect of MgATP on assembly.J. Muscle Res. Cell Motil. 7, 413–420.
Cohen C., Lanar D. E., and Parry D. A. D. (1987) Amino acid sequence and structural repeats in schistosome paramyosin match those of myosin.Biosci. Rep. 7, 11–16.
Cohen C., Szent-Györgyi A. G., and Kendrick-Jones J. (1971) Paramyosin and the filaments of molluscan “catch” muscles. I. Paramyosin: structure and assembly.J. Mol. Biol. 56, 223–237.
Coluccio L. M. and Bretscher A. (1987) Calcium-regulated cooperative binding of the microvillar 110K-calmodulin complex to F-actin: formation of decorated filaments.J. Cell Biol. 105, 325–333.
Conzelman K. A. and Mooseker, M. S. (1987) The 100 kD protein-calmodulin complex of the intestinal microvillus is an actin-activated MyATPase.J. Cell Biol. 105, 313–324.
Cooper J. and Trinick J. (1984) Binding and location of AMP deaminase in rabbit psoas muscle myofibrils.J. Mol. Biol. 177, 137–152.
Coulson A., Sulston J., Brenner S., and Karn J. (1986) Towards a physical map of the genome of the nematodeCaenorhabditis elegans.Proc. Natl. Acad. Sci. USA 83, 7821–7825.
Craig R. and Megerman J. (1977) Assembly of smooth muscle myosin into side-polar filaments.J. Cell Biol. 75, 990–996.
Craig R. and Offer G. (1976) The location of C-protein in rabbit skeletal muscle.Proc. Roy. Soc. Lond. B. 192, 451–464.
Cummins, C. and Anderson, P. (1988) Regulatory myosin light-chain genes ofCaenorhabditis elegans.Mol. Cell. Biol. 8, 5339–5349.
Davis J. S. (1988) Assembly processes in vertebrate skeletal thick filament formation.Annu. Rev. Biophys. Chem. 17, 217–235.
De Lozanne A. and Spudich J. A. (1987) Disruption of theDictyostelium myosin heavy chain gene by homologous recombination.Science 236, 1086–1091.
Dibb N. J., Brown D. M., Karn J., Moerman D. G., Bolten S. L., and Waterston R. H. (1985) Sequence analysis of mutations that affect the synthesis, assembly and enzymatic activity of theunc-54 myosin heavy chain ofCaenorhabditis elegans.J. Mol. Biol. 183, 543–551.
Dibb N. J., Maruyama I. N., Krause M., and Karn J. (1989) Sequence analysis of the completeCaenorhabditis elegans myosin heavy chain gene family.J. Mol. Biol. 205, 603–613.
Eide D. and Anderson P. (1985) The gene structures of spontaneous mutations affecting aCaenorhabditis elegans myosin heavy chain gene.Genetics 109, 67–79.
Emerson C. P. and Bernstein S. I. (1987) Molecular genetics of myosin.Annu. Rev. Biochem. 56, 695–726.
Epstein H. F. (1986) Differential roles of myosin isoforms in filament assembly.UCLA Symp. Mol. Cell. Biol. New Ser. 29, 653–666.
Epstein H. F. (1988a) Modulation of myosin assembly.UCLA Symp. Mol. Cell Biol. New Ser. 93, 207–219.
Epstein H. F. (1988b) Modulation of myosin assembly.Bioessays 9, 197–200.
Epstein H. F. and Thomson J. N. (1974) Temperature sensitive mutation affecting myofilament assembly inCaenorhabditis elegans.Nature 250, 579–580.
Epstein H. F., Berliner G. C., Casey D. L., and Ortiz I. (1988) Purified thick filaments from the nematodeCaenorhabditis elegans: evidence for multiple proteins associated with core structures.J. Cell Biol. 106, 1985–1995.
Epstein H. F., Berman S. A., and Miller D. M. III (1982a) Myosin synthesis and assembly in nematode body-wall muscle, inMuscle Development: Molecular and Cellular Control, Pearson M. and Epstein H. F., eds., Cold Spring Harbor Laboratory, Cold Spring Harbor, New York, pp. 7–14.
Epstein H. F., Miller D. M., III, Ortiz I., and Berliner, G. C. (1985) Myosin and paramyosin are organized about a newly identified core structure.J. Cell Biol. 100, 904–915.
Epstein H. F., Ortiz I., and Berliner G. C. (1987) Assemblages of multiple thick filaments in nematode mutants.J. Muscle Res. Cell. Motil. 8, 527–536.
Epstein H. F., Ortiz I., and Mackinnon L. A. T. (1986) The alteration of myosin isoform compartmentation in specific mutants ofCaenorhabditis elegans.J. Cell. Biol. 103, 985–993.
Epstein H. F., Waterston R. H., and Brenner S. (1974) A mutant affecting the heavy chain of myosin inCaenorhabditis elegans.J. Mol. Biol. 90, 291–300.
Gadasi H. and Korn E. D. (1980) Evidence for differential intracellular localization of the Acanthamoeba myosin isoenzymes.Nature 286, 452–456.
Garcea R. L., Schachat F., and Epstein H. F. (1978) Coordinate synthesis of two myosins in wild-type and mutant nematode muscle during larval development.Cell 15, 421–428.
Geisselsoder J., Chidambaram M., and Goldstein R. (1978) Transcriptional control of capsid size in the P2:P4 bacteriophage system.J. Mol. Biol. 126, 447–456.
Goldberg A. F. and Chang T. W. (1978) Regulation and significance of amino acid metabolism in skeletal muscle.Fed. Proc. 37, 2301–2307.
Goldberg A. L., Griffin G. E., and Dice J. F. (1977) Regulation of protein turnover in normal and dystrophic muscle, inPathogenesis of Human Muscular Dystrophies, Rowland L. P., ed., Excerpta Medica, Amsterdam, pp. 376–385.
Gossett L. A., Hecht R. M., and Epstein H. F. (1982) Muscle differentiation in normal and cleavage-arrested mutant embryos ofCaenorhabditis elegans.Cell 30, 193–204.
Grove B. K., Cerny L., Perriard J-C., and Eppenberger H. M. (1985) Myomesin and M-protein: expression of two M-band proteins in pectoral muscle and heart during development.J. Cell Biol. 101, 1413–1421.
Harrington W. F., Burke, M., and Barton J. S. (1972) Association of myosin to form contractile systems.Cold Spring Harbor Symp. Quart. Biol. 37, 77–85.
Harris H. E. and Epstein H. F. (1977) Myosin and paramyosin ofCaenorhabditis elegans: Biochemical and structural properties of wild-type and mutant proteins.Cell 10, 709–719.
Honda S. and Epstein H. F. (1990) Modulation of muscle gene expression inCaenorhabditis elegans: Differential regulation of transcripts, mRNAs, and polypeptides for thick filament proteins during nematode development.Proc. Natl. Acad. Sci. USA 87, 876–880.
Huszar G. (1972) Amino acid sequences around the two ε-N-trimethyllysine residues in rabbit skeletal muscle myosin.J. Biol. Chem. 247, 4057–4062.
Huszar G. and Elzinga M. (1973) Homologous methylated and nonmethylated histidine peptides in skeletal and cardiac myosins.J. Biol. Chem. 247, 745–753.
Huxley H. E. (1963) Electron microscopy studies of native and synthetic protein filaments from striated muscle.J. Mol. Biol. 7, 281–308.
Huxley H. E. (1969) The mechanism of muscular contraction.Science 164, 1356–1366.
Jorgensen A. O., Kalinins V. I., Zubryzycka E., and MacLennan D. H. (1977) Assembly of the sarcoplasmic reticulum.J. Cell Biol. 74, 287–298.
Kagawa H., Gengyo K., McLachlan A. D., Brenner S., and Karn J. (1989) The paramyosin gene (unc-15) ofCaenorhabditis elegans: Molecular cloning, nucleotide sequence and models for thick fila- ment assembly.J. Mol. Biol. 207, 311–333.
Karn J., Dibb N. J., and Miller D. M. (1985) Cloning nematode myosin genes, inCell and Muscle Motility, Shay J., ed., vol. 6, Plenum Press, New York, pp. 185–237.
Karn J., McLachlan A. D., and Barnett L. (1982),Unc-54 myosin heavy-chain gene ofCaenorhabditis elegans: Genetics, sequence, structure, inMuscle Development: Molecular and Cellular Control, Pearson M. and Epstein H. F., eds., Cold Spring Harbor Laboratory, Cold Spring Harbor, New York, pp. 129–142.
Katsura I. (1987) Determination of bacteriophage 1 tail length by a protein ruler.Nature 327, 73–75.
Kensler R. W. and Stewart M. (1983) Frog skeletal muscle thick filaments are three-stranded.J. Cell Biol. 96, 1797–1802.
Knecht D. A. and Loomis W. F. (1987) Antisense RNA inactivation of myosin heavy chain gene expression inDictyostelium discoideum.Science 236, 1081–1086.
Kordeli E., Cartaud J., Nghiem H-O., Devilliers-Thiérry A., and Changeux J-P. (1989) Asynchronous assembly of the acetylcholine receptor and of the 43-kDv, protein in the postsynaptic membrane of developingTorpedo marmorata electrocyte.J. Cell Biol. 108, 127–139.
Kuczmarski E. R. and Spudich J. A. (1980) Regulation of myosin self-assembly: phosphorylation ofDictyostelium heavy chain inhibits formation of thick filaments.Proc. Natl. Acad. Sci. USA 71, 7292–7296.
Levine R. J. C., Elfvin M., Dewey M. M., and Walcott B. (1976) Paramyosin in invertebrate muscles II. Contentin relation to structure and function.J. Cell Biol. 70, 273–279.
Lowey S., Slayter H. S., Weeds A., and Baker H. (1969) Substructure of the myosin molecule. I. Subfragments of myosin by enzymic digestion.J. Mol. Biol. 42, 1–29.
Mackenzie J. M., Jr. and Epstein H. F. (1980) Paramyosin is necessary for determination of nematode thick filament in vivo.Cell 22, 747–755.
Mackenzie J. M., Jr. and Epstein, H. F. (1981) Electron microscopy of nematode thick filaments.J. Ultrastruct. Res. 76, 277–285.
Mackenzie J. M., Jr. Schachat F., and Epstein H. F. (1978a) Immunocytochemical localization of two myosins within the same muscle cells inCaenorhabditis elegans.Cell 15, 413–420.
Mackenzie J. M., Jr. Garcea R. L., Zengel J. M., and Epstein H. F. (1978b) Muscle development inCaenorhabditis elegans mutants exhibiting retarded sarcomere construction.Cell 15, 751–762.
MacLeod A. R., Karn J., and Brenner S. (1981) Molecular analysis of theunc-54 myosin heavy chain gene ofCaenorhabditis elegans.Nature 291, 386–390.
MacLeod A. R., Waterston R. H., and Brenner S. (1977a) An internal deletion mutant of a myosin heavy chain inCaenorhabditis elegans.Proc. Natl. Acad. Sci. USA 74, 5336–5340.
MacLeod A. R., Waterston R. H., Fishpool R. M., and Brenner S. (1977b) Identification of the structural genes for a myosin heavy-chain inCaenorhabditis elegans.J. Mol. Biol. 14, 133–140.
Masaki T. and Takaiti O. (1974) M-protein.J. Biochem. (Tokyo)75, 367–380.
McLachlan A. D. and Karn J. (1983) Periodic features in the amino acid sequence of nematode myosin rod.J. Mol. Biol. 164, 605–626.
Merlie J. P. and Lindstrom J. (1983) Assembly in vivo of mouse muscle acetylcholine receptor: identification of an a subunit species that may be an assembly intermediate.Cell 34, 747–757.
Miller D. M. and Maruyama I. (1986) Thesup-3 locus is closely linked to a myosin heavy chain gene inCaenorhabditis elegans.UCLA Symp. Mol. Cell Biol. New Ser. 29, 629–638.
Miller D. M. III, Ortiz I., Berliner G. C., and Epstein H. F. (1983) Differential localization of two myosins within nematode thick filaments.Cell 34, 477–490.
Miller D. M., Stockdale F. E., and Karn J. (1986) Immunological identification of the genes encoding the four myosin heavy chain isoforms ofCaenorhabditis elegans.Proc. Natl. Acad. Sci. 83, 2305–2309.
Moerman D. G., Benian G. M., and Waterston R. H. (1986) Molecular cloning of the muscle geneunc-22 inCaenorhabditis elegans by Tcl transposon tagging.Proc. Natl. Acad. Sci. USA 83, 2579–2583.
Moerman D. G., Benian G. M., Barstead R. J., Schreifer L., and Waterston R. H. (1988) Identification and intracellular localization of theunc-22 gene product ofCaenorhabditis elegans.Genes Dev. 2, 93–105.
Moerman D. G., Plurad S. Waterston R. H., and Baillie D. L. (1982) Mutations in theunc-54 myosin heavy chain gene ofCaenorhabditis elegans that alter contractility but not muscle structure.Cell 29, 773–781.
Morimoto K. and Harrington W. F. (1973) Isolation and composition of thick filaments from rabbit skeletal muscle.J. Mol. Biol. 77, 165–175.
Niederman R. and Peters L. K. (1982) Native bare zone assemblage nucleates myosin filament assembly.J. Mol. Biol. 161, 505–517.
O’Donnell P. T. and Bernstein S. J. (1988) Molecular and ultrastructural defects in aDrosophila myosin heavy chain mutant in: differential effects on muscle function produced by similar thick filament abnormalities.J. Cell Biol. 107, 2601–2612.
Otsuka A. J. (1986)sup-3 Suppression affects muscle structure and myosin heavy chain accumulation inCaenorhabditis elegans.UCLA Symp. Mol. Cell. Biol. New Ser. 29, 619–628.
Pollard T. D. and Korn E. D. (1973) Acanthamoeba myosin I.J. Biol. Chem. 248, 4682–4690.
Reisler E., Smith C., and Seegan G. (1980) Myosin minifilaments.J. Mol. Biol. 143, 129–145.
Riddle D. L. and Brenner S. (1978) Indirect suppression inCaenorhabditis elegans.Genetics 89, 299–314.
Saad A. D., Pardee J. D., and Fischman D. A. (1987) Dynamic exchange of myosin molecules between thick filaments.Proc. Natl. Acad. Sci. USA 83, 9483–9487.
Schachat, F., Garcea R. L., and Epstein H. F. (1978) Myosins exist as homodimers of heavy chains: demonstration with specific antibody purified by nematode mutant myosin affinity chromatography.Cell 15, 405–411.
Schachat F., Harris H. E., and Epstein H. F. (1977) Two homogeneous myosins in body-wall muscle ofCaenorhabditis elegans.Cell 10, 721–728.
Scholey J. M., Taylor K. A., and Kendrick-Jones J. (1980) Regulation of non-muscle assembly by calmodulin-dependent light chain kinase.Nature 287, 233–235.
Schreifer L. and Waterston R. H. (1989) Phosphorylation of the N-terminal region ofCaenorhabditis elegans paramyosin.J. Mol. Biol. 207, 451–454.
Showe K. and Onorato L. (1978) Kinetic factors and form determination of the head of bacteriophage T4.Proc. Natl. Acad. Sci. USA 75 4165–4169.
Starr R. and Offer G. (1973) Polarity of the myosin molecule.J. Mol. Biol. 81, 17–31.
Strehler E. E., Pelloni G., Heizmann C. W. and Eppenberger H. M. (1980) Biochemical and ultrastructural aspects of M 165,000 M-protein in cross-striated chicken muscle.J. Cell Biol. 86, 775–783.
Sulston J. E. and Brenner S. (1974) The DNA ofCaenorhabditis elegans.Genetics 77, 95–104.
Szent-Györgyi A. G., Cohen C., and Kendrick-Jones J. (1971) Paramyosin and the filaments of molluscan “catch muscles.” II. Native filaments: isolation and characterization.J. Mol. Biol. 56, 239–258.
Taurmino J., Wolitzky B. A., Takeyasa K., Tamkun M. M., and Fambrough D. M. (1989) Up-regulation of the sodium pump in primary chick muscle cultures.UCLA Symp. Mol. Cell Biol. New Ser. 93, 327–334.
Taylor L. D. and Bandman E. (1989) Distribution of fast myosin heavy chain isoforms in thick filaments of developing chicken pectoral muscle.J. Cell Biol. 108, 533–542.
Trybus K. M. and Lowey S. (1987) Assembly of smooth muscle myosin minifilaments: effects of phosphorylation and nucleotide binding.J. Cell. Biol. 105, 3007–3014.
Turner D. C., Walliman T., and Eppenberger H. M. (1973) A protein that binds specifically to the M-line of skeletal muscle is identified as the muscle form of creatine kinase.Proc. Natl. Acad. Sci. USA 70, 702–705.
Warrick H. M. and Spudich J. A. (1987) Myosin: structure and function in cell motility.Ann. Rev. Cell Biol. 3, 379–422.
Waterston R. H. (1988) Muscle, inThe Nematode Caenorhabditis elegans, W. B. Wood, ed., Cold Spring Harbor Laboratory, Cold Spring Harbor New York, pp. 281–335.
Waterston R. H. (1989) The minor myosin heavy chain, mhcA, ofCaenorhabditis elegans is necessary for the initiation of thick filament assembly.EMBO J. 8, 3429–3436.
Waterson R. H., Epstein H. F., and Brenner S. (1974) Paramyosin inCaenorhabditis elegans.J. Mol. Biol. 90, 285–290.
Waterston R. H., Thomson J. N., and Brenner S. (1980) Mutants with altered muscle structure inCaenorhabditis elegans.Dev. Biol. 77, 271–302.
Waterston R. H., Fishpool R. M., and Brenner S. (1977) Mutants affecting paramyosin inCaenorhabditis elegans.J. Mol. Biol. 117, 679–697.
Waterston R. H. and Brenner S. (1978) A suppressor mutation in the nematode acting on specific alleles of many genes.Nature 275, 715–719.
Waterston R. H., Moerman D. G., Baillie D. L., and Lane T. R. (1982) Mutations affecting myosin heavy chain accumulation and function in the nematodeCaenorhabditis elegans, inDisorders of the Motor Unit, Schotland D. M., ed., John Wiley, New York, pp. 747–760.
Wenderoth M. P. and Eisenberg B. R. (1987) Heterogeneous distribution of nascent heavy chains into thick filaments of cardiac myocytes in thyroid treated rabbits.J. Cell. Biol. 105, 2971–2780.
Whalen R. G., Butler-Browne G. S., Pinset C., Toutant M., Watkins S. C., Ajioka J., Laurent C., McCormick D., and Riley G. P. (1986) Control of myosin isoform expression during skeletal muscle development in rodents.UCLA Symp. Mol. Cell. Biol., New Ser. 29, 237–251.
Wray J. (1979) Structure of the backbone in myosin filaments of muscle.Nature 277, 37–40
Zengel J. M. and Epstein H. F. (1980a) Identification of genetic elements associated with muscle structure in the nematodeCaenorhabditis elegans.Cell Motil. 1, 73–97.
Zengel J. M. and Epstein H. F. (1980b) Mutants altering coordinate synthesis of specific myosins during nematode muscle development.Proc. Natl. Acad. Sci. USA 77, 852–856.
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Epstein, H.F. Genetic analysis of myosin assembly inCaenorhabditis elegans . Mol Neurobiol 4, 1–25 (1990). https://doi.org/10.1007/BF02935583
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DOI: https://doi.org/10.1007/BF02935583