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. 1990 Jun 1;110(6):1999–2011. doi: 10.1083/jcb.110.6.1999

Molecular genetics of Drosophila alpha-actinin: mutant alleles disrupt Z disc integrity and muscle insertions

PMCID: PMC2116134  PMID: 2112549

Abstract

We have isolated a Drosophila melanogaster alpha-actinin gene and partially characterized several mutant alleles. The Drosophila protein sequence is very similar (68% identity) to those of chicken alpha- actinin isoforms, but less closely related (30% identity) to Dictyostelium alpha-actinin. The gene is within subdivision 2C of the X chromosome, coincident with 15 lethal (1)2Cb mutations. At least four alleles, l(1)2Cb1, l(1)2Cb2, l(1)2Cb4, and l(1)2Cb5 are interrupted by rearrangement breakpoints and must be null. In all four cases, hemizygous mutants complete embryogenesis and do not die until the second day of larval growth, signifying that either the role of alpha- actinin in nonmuscle cells is redundant or that a distinct and only distantly related gene encodes the non-muscle isoform. Allelic but less severely affected fliA mutants are apparently due to point mutations, and develop into adults having thoracic muscle abnormalities. EM of mutant muscles reveals that Z discs and myofibrillar attachments are disrupted, whereas epithelial "tendon" cells are less affected. We discuss these phenotypes in the light of presumed in vivo alpha-actinin functions.

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Selected References

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  1. Arimura C., Suzuki T., Yanagisawa M., Imamura M., Hamada Y., Masaki T. Primary structure of chicken skeletal muscle and fibroblast alpha-actinins deduced from cDNA sequences. Eur J Biochem. 1988 Nov 15;177(3):649–655. doi: 10.1111/j.1432-1033.1988.tb14419.x. [DOI] [PubMed] [Google Scholar]
  2. Baron M. D., Davison M. D., Jones P., Critchley D. R. The sequence of chick alpha-actinin reveals homologies to spectrin and calmodulin. J Biol Chem. 1987 Dec 25;262(36):17623–17629. [PubMed] [Google Scholar]
  3. Baron M. D., Davison M. D., Jones P., Patel B., Critchley D. R. Isolation and characterization of a cDNA encoding a chick alpha-actinin. J Biol Chem. 1987 Feb 25;262(6):2558–2561. [PubMed] [Google Scholar]
  4. Beall C. J., Sepanski M. A., Fyrberg E. A. Genetic dissection of Drosophila myofibril formation: effects of actin and myosin heavy chain null alleles. Genes Dev. 1989 Feb;3(2):131–140. doi: 10.1101/gad.3.2.131. [DOI] [PubMed] [Google Scholar]
  5. Benton W. D., Davis R. W. Screening lambdagt recombinant clones by hybridization to single plaques in situ. Science. 1977 Apr 8;196(4286):180–182. doi: 10.1126/science.322279. [DOI] [PubMed] [Google Scholar]
  6. Blanchard A., Ohanian V., Critchley D. The structure and function of alpha-actinin. J Muscle Res Cell Motil. 1989 Aug;10(4):280–289. doi: 10.1007/BF01758424. [DOI] [PubMed] [Google Scholar]
  7. Burridge K., Feramisco J. R. Non-muscle alpha actinins are calcium-sensitive actin-binding proteins. Nature. 1981 Dec 10;294(5841):565–567. doi: 10.1038/294565a0. [DOI] [PubMed] [Google Scholar]
  8. Byers T. J., Husain-Chishti A., Dubreuil R. R., Branton D., Goldstein L. S. Sequence similarity of the amino-terminal domain of Drosophila beta spectrin to alpha actinin and dystrophin. J Cell Biol. 1989 Oct;109(4 Pt 1):1633–1641. doi: 10.1083/jcb.109.4.1633. [DOI] [PMC free article] [PubMed] [Google Scholar]
  9. Caveney S. Muscle attachment related to cuticle architecture in Apterygota. J Cell Sci. 1969 Mar;4(2):541–559. doi: 10.1242/jcs.4.2.541. [DOI] [PubMed] [Google Scholar]
  10. 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]
  11. Cheng N. Q., Deatherage J. F. Three-dimensional reconstruction of the Z disk of sectioned bee flight muscle. J Cell Biol. 1989 May;108(5):1761–1774. doi: 10.1083/jcb.108.5.1761. [DOI] [PMC free article] [PubMed] [Google Scholar]
  12. Deatherage J. F., Cheng N. Q., Bullard B. Arrangement of filaments and cross-links in the bee flight muscle Z disk by image analysis of oblique sections. J Cell Biol. 1989 May;108(5):1775–1782. doi: 10.1083/jcb.108.5.1775. [DOI] [PMC free article] [PubMed] [Google Scholar]
  13. Duhaiman A. S., Bamburg J. R. Isolation of brain alpha-actinin. Its characterization and a comparison of its properties with those of muscle alpha-actinins. Biochemistry. 1984 Apr 10;23(8):1600–1608. doi: 10.1021/bi00303a003. [DOI] [PubMed] [Google Scholar]
  14. Endo T., Masaki T. Molecular properties and functions in vitro of chicken smooth-muscle alpha-actinin in comparison with those of striated-muscle alpha-actinins. J Biochem. 1982 Nov;92(5):1457–1468. doi: 10.1093/oxfordjournals.jbchem.a134070. [DOI] [PubMed] [Google Scholar]
  15. Geiger B., Tokuyasu K. T., Singer S. J. Immunocytochemical localization of alpha-actinin in intestinal epithelial cells. Proc Natl Acad Sci U S A. 1979 Jun;76(6):2833–2837. doi: 10.1073/pnas.76.6.2833. [DOI] [PMC free article] [PubMed] [Google Scholar]
  16. Goldspink G. Sarcomere length during post-natal growth of mammalian muscle fibres. J Cell Sci. 1968 Dec;3(4):539–548. doi: 10.1242/jcs.3.4.539. [DOI] [PubMed] [Google Scholar]
  17. Goldstein M. A., Schroeter J. P., Sass R. L. The Z-band lattice in a slow skeletal muscle. J Muscle Res Cell Motil. 1982 Sep;3(3):333–348. doi: 10.1007/BF00713041. [DOI] [PubMed] [Google Scholar]
  18. Homyk T., Jr, Emerson C. P., Jr Functional interactions between unlinked muscle genes within haploinsufficient regions of the Drosophila genome. Genetics. 1988 May;119(1):105–121. doi: 10.1093/genetics/119.1.105. [DOI] [PMC free article] [PubMed] [Google Scholar]
  19. Imamura M., Endo T., Kuroda M., Tanaka T., Masaki T. Substructure and higher structure of chicken smooth muscle alpha-actinin molecule. J Biol Chem. 1988 Jun 5;263(16):7800–7805. [PubMed] [Google Scholar]
  20. Karlik C. C., Mahaffey J. W., Coutu M. D., Fyrberg E. A. Organization of contractile protein genes within the 88F subdivision of the D. melanogaster third chromosome. Cell. 1984 Jun;37(2):469–481. doi: 10.1016/0092-8674(84)90377-5. [DOI] [PubMed] [Google Scholar]
  21. Koenig M., Monaco A. P., Kunkel L. M. The complete sequence of dystrophin predicts a rod-shaped cytoskeletal protein. Cell. 1988 Apr 22;53(2):219–228. doi: 10.1016/0092-8674(88)90383-2. [DOI] [PubMed] [Google Scholar]
  22. Kretsinger R. H. Crystallographic studies of calmodulin and homologs. Ann N Y Acad Sci. 1980;356:14–19. doi: 10.1111/j.1749-6632.1980.tb29594.x. [DOI] [PubMed] [Google Scholar]
  23. Lai-Fook J. The structure of developing muscle insertions in insects. J Morphol. 1967 Dec;123(4):503–527. doi: 10.1002/jmor.1051230411. [DOI] [PubMed] [Google Scholar]
  24. 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]
  25. Maniatis T., Hardison R. C., Lacy E., Lauer J., O'Connell C., Quon D., Sim G. K., Efstratiadis A. The isolation of structural genes from libraries of eucaryotic DNA. Cell. 1978 Oct;15(2):687–701. doi: 10.1016/0092-8674(78)90036-3. [DOI] [PubMed] [Google Scholar]
  26. 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]
  27. Maxam A. M., Gilbert W. Sequencing end-labeled DNA with base-specific chemical cleavages. Methods Enzymol. 1980;65(1):499–560. doi: 10.1016/s0076-6879(80)65059-9. [DOI] [PubMed] [Google Scholar]
  28. Mimura N., Asano A. Further characterization of a conserved actin-binding 27-kDa fragment of actinogelin and alpha-actinins and mapping of their binding sites on the actin molecule by chemical cross-linking. J Biol Chem. 1987 Apr 5;262(10):4717–4723. [PubMed] [Google Scholar]
  29. Noegel A. A., Witke W. Inactivation of the alpha-actinin gene in Dictyostelium. Dev Genet. 1988;9(4-5):531–538. doi: 10.1002/dvg.1020090429. [DOI] [PubMed] [Google Scholar]
  30. Noegel A., Witke W., Schleicher M. Calcium-sensitive non-muscle alpha-actinin contains EF-hand structures and highly conserved regions. FEBS Lett. 1987 Sep 14;221(2):391–396. doi: 10.1016/0014-5793(87)80962-6. [DOI] [PubMed] [Google Scholar]
  31. Perrimon N., Engstrom L., Mahowald A. P. Developmental genetics of the 2C-D region of the Drosophila X chromosome. Genetics. 1985 Sep;111(1):23–41. doi: 10.1093/genetics/111.1.23. [DOI] [PMC free article] [PubMed] [Google Scholar]
  32. Reedy M. C., Beall C., Fyrberg E. Formation of reverse rigor chevrons by myosin heads. Nature. 1989 Jun 8;339(6224):481–483. doi: 10.1038/339481a0. [DOI] [PubMed] [Google Scholar]
  33. Reedy M. K., Reedy M. C. Rigor crossbridge structure in tilted single filament layers and flared-X formations from insect flight muscle. J Mol Biol. 1985 Sep 5;185(1):145–176. doi: 10.1016/0022-2836(85)90188-3. [DOI] [PubMed] [Google Scholar]
  34. Reedy M. K. Ultrastructure of insect flight muscle. I. Screw sense and structural grouping in the rigor cross-bridge lattice. J Mol Biol. 1968 Jan 28;31(2):155–176. doi: 10.1016/0022-2836(68)90437-3. [DOI] [PubMed] [Google Scholar]
  35. Sanger F., Nicklen S., Coulson A. R. DNA sequencing with chain-terminating inhibitors. Proc Natl Acad Sci U S A. 1977 Dec;74(12):5463–5467. doi: 10.1073/pnas.74.12.5463. [DOI] [PMC free article] [PubMed] [Google Scholar]
  36. Sato M., Schwarz W. H., Pollard T. D. Dependence of the mechanical properties of actin/alpha-actinin gels on deformation rate. 1987 Feb 26-Mar 4Nature. 325(6107):828–830. doi: 10.1038/325828a0. [DOI] [PubMed] [Google Scholar]
  37. Schleicher M., Noegel A., Schwarz T., Wallraff E., Brink M., Faix J., Gerisch G., Isenberg G. A Dictyostelium mutant with severe defects in alpha-actinin: its characterization using cDNA probes and monoclonal antibodies. J Cell Sci. 1988 May;90(Pt 1):59–71. doi: 10.1242/jcs.90.1.59. [DOI] [PubMed] [Google Scholar]
  38. Smith S. J. Neuronal cytomechanics: the actin-based motility of growth cones. Science. 1988 Nov 4;242(4879):708–715. doi: 10.1126/science.3055292. [DOI] [PubMed] [Google Scholar]
  39. 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]
  40. Wallraff E., Schleicher M., Modersitzki M., Rieger D., Isenberg G., Gerisch G. Selection of Dictyostelium mutants defective in cytoskeletal proteins: use of an antibody that binds to the ends of alpha-actinin rods. EMBO J. 1986 Jan;5(1):61–67. doi: 10.1002/j.1460-2075.1986.tb04178.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  41. Wensink P. C., Finnegan D. J., Donelson J. E., Hogness D. S. A system for mapping DNA sequences in the chromosomes of Drosophila melanogaster. Cell. 1974 Dec;3(4):315–325. doi: 10.1016/0092-8674(74)90045-2. [DOI] [PubMed] [Google Scholar]
  42. Yamaguchi M., Izumimoto M., Robson R. M., Stromer M. H. Fine structure of wide and narrow vertebrate muscle Z-lines. A proposed model and computer simulation of Z-line architecture. J Mol Biol. 1985 Aug 20;184(4):621–643. doi: 10.1016/0022-2836(85)90308-0. [DOI] [PubMed] [Google Scholar]
  43. Young R. A., Davis R. W. Yeast RNA polymerase II genes: isolation with antibody probes. Science. 1983 Nov 18;222(4625):778–782. doi: 10.1126/science.6356359. [DOI] [PubMed] [Google Scholar]

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