Skip to main content
Genetics logoLink to Genetics
. 1994 May;137(1):151–164. doi: 10.1093/genetics/137.1.151

Recovery of Dominant, Autosomal Flightless Mutants of Drosophila Melanogaster and Identification of a New Gene Required for Normal Muscle Structure and Function

R M Cripps 1, E Ball 1, M Stark 1, A Lawn 1, J C Sparrow 1
PMCID: PMC1205932  PMID: 8056306

Abstract

To identify further mutations affecting muscle function and development in Drosophila melanogaster we recovered 22 autosomal dominant flightless mutations. From these we have isolated eight viable and lethal alleles of the muscle myosin heavy chain gene, and seven viable alleles of the indirect flight muscle (IFM)-specific Act88F actin gene. The Mhc mutations display a variety of phenotypic effects, ranging from reductions in myosin heavy chain content in the indirect flight muscles only, to reductions in the levels of this protein in other muscles. The Act88F mutations range from those which produce no stable actin and have severely abnormal myofibrillar structure, to those which accumulate apparently normal levels of actin in the flight muscles but which still have abnormal myofibrils and fly very poorly. We also recovered two recessive flightless mutants on the third chromosome. The remaining five dominant flightless mutations are all lethal alleles of a gene named lethal(3)Laker. The Laker alleles have been characterized and the gene located in polytene bands 62A10,B1-62B2,4. Laker is a previously unidentified locus which is haplo-insufficient for flight. In addition, adult wild-type heterozygotes and the lethal larval trans-heterozygotes show abnormalities of muscle structure indicating that the Laker gene product is an important component of muscle.

Full Text

The Full Text of this article is available as a PDF (5.5 MB).

Selected References

These references are in PubMed. This may not be the complete list of references from this article.

  1. Ayme-Southgate A., Vigoreaux J., Benian G., Pardue M. L. Drosophila has a twitchin/titin-related gene that appears to encode projectin. Proc Natl Acad Sci U S A. 1991 Sep 15;88(18):7973–7977. doi: 10.1073/pnas.88.18.7973. [DOI] [PMC free article] [PubMed] [Google Scholar]
  2. Ball E., Karlik C. C., Beall C. J., Saville D. L., Sparrow J. C., Bullard B., Fyrberg E. A. Arthrin, a myofibrillar protein of insect flight muscle, is an actin-ubiquitin conjugate. Cell. 1987 Oct 23;51(2):221–228. doi: 10.1016/0092-8674(87)90149-8. [DOI] [PubMed] [Google Scholar]
  3. Barbas J. A., Galceran J., Krah-Jentgens I., de la Pompa J. L., Canal I., Pongs O., Ferrús A. Troponin I is encoded in the haplolethal region of the Shaker gene complex of Drosophila. Genes Dev. 1991 Jan;5(1):132–140. doi: 10.1101/gad.5.1.132. [DOI] [PubMed] [Google Scholar]
  4. Bautch V. L., Storti R. V., Mischke D., Pardue M. L. Organization and expression of Drosophila tropomyosin genes. J Mol Biol. 1982 Dec 5;162(2):231–250. doi: 10.1016/0022-2836(82)90524-1. [DOI] [PubMed] [Google Scholar]
  5. Beall C. J., Fyrberg E. Muscle abnormalities in Drosophila melanogaster heldup mutants are caused by missing or aberrant troponin-I isoforms. J Cell Biol. 1991 Sep;114(5):941–951. doi: 10.1083/jcb.114.5.941. [DOI] [PMC free article] [PubMed] [Google Scholar]
  6. 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]
  7. 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]
  8. Bellen H. J., O'Kane C. J., Wilson C., Grossniklaus U., Pearson R. K., Gehring W. J. P-element-mediated enhancer detection: a versatile method to study development in Drosophila. Genes Dev. 1989 Sep;3(9):1288–1300. doi: 10.1101/gad.3.9.1288. [DOI] [PubMed] [Google Scholar]
  9. Bernstein S. I., Hansen C. J., Becker K. D., Wassenberg D. R., 2nd, Roche E. S., Donady J. J., Emerson C. P., Jr Alternative RNA splicing generates transcripts encoding a thorax-specific isoform of Drosophila melanogaster myosin heavy chain. Mol Cell Biol. 1986 Jul;6(7):2511–2519. doi: 10.1128/mcb.6.7.2511. [DOI] [PMC free article] [PubMed] [Google Scholar]
  10. Bernstein S. I., Mogami K., Donady J. J., Emerson C. P., Jr Drosophila muscle myosin heavy chain encoded by a single gene in a cluster of muscle mutations. 1983 Mar 31-Apr 6Nature. 302(5907):393–397. doi: 10.1038/302393a0. [DOI] [PubMed] [Google Scholar]
  11. Bernstein S. I., O'Donnell P. T., Cripps R. M. Molecular genetic analysis of muscle development, structure, and function in Drosophila. Int Rev Cytol. 1993;143:63–152. doi: 10.1016/s0074-7696(08)61874-4. [DOI] [PubMed] [Google Scholar]
  12. Brunk B. P., Martin E. C., Adler P. N. Molecular genetics of the Posterior sex combs/Suppressor 2 of zeste region of Drosophila: aberrant expression of the Suppressor 2 of zeste gene results in abnormal bristle development. Genetics. 1991 May;128(1):119–132. doi: 10.1093/genetics/128.1.119. [DOI] [PMC free article] [PubMed] [Google Scholar]
  13. Bullard B., Bell J., Craig R., Leonard K. Arthrin: a new actin-like protein in insect flight muscle. J Mol Biol. 1985 Apr 5;182(3):443–454. doi: 10.1016/0022-2836(85)90203-7. [DOI] [PubMed] [Google Scholar]
  14. Byers T. J., Dubreuil R., Branton D., Kiehart D. P., Goldstein L. S. Drosophila spectrin. II. Conserved features of the alpha-subunit are revealed by analysis of cDNA clones and fusion proteins. J Cell Biol. 1987 Nov;105(5):2103–2110. doi: 10.1083/jcb.105.5.2103. [DOI] [PMC free article] [PubMed] [Google Scholar]
  15. Chun M., Falkenthal S. Ifm(2)2 is a myosin heavy chain allele that disrupts myofibrillar assembly only in the indirect flight muscle of Drosophila melanogaster. J Cell Biol. 1988 Dec;107(6 Pt 2):2613–2621. doi: 10.1083/jcb.107.6.2613. [DOI] [PMC free article] [PubMed] [Google Scholar]
  16. Collier V. L., Kronert W. A., O'Donnell P. T., Edwards K. A., Bernstein S. I. Alternative myosin hinge regions are utilized in a tissue-specific fashion that correlates with muscle contraction speed. Genes Dev. 1990 Jun;4(6):885–895. doi: 10.1101/gad.4.6.885. [DOI] [PubMed] [Google Scholar]
  17. Drummond D. R., Hennessey E. S., Sparrow J. C. Characterisation of missense mutations in the Act88F gene of Drosophila melanogaster. Mol Gen Genet. 1991 Apr;226(1-2):70–80. doi: 10.1007/BF00273589. [DOI] [PubMed] [Google Scholar]
  18. Drummond D. R., Peckham M., Sparrow J. C., White D. C. Alteration in crossbridge kinetics caused by mutations in actin. Nature. 1990 Nov 29;348(6300):440–442. doi: 10.1038/348440a0. [DOI] [PubMed] [Google Scholar]
  19. Falkenthal S., Parker V. P., Mattox W. W., Davidson N. Drosophila melanogaster has only one myosin alkali light-chain gene which encodes a protein with considerable amino acid sequence homology to chicken myosin alkali light chains. Mol Cell Biol. 1984 May;4(5):956–965. doi: 10.1128/mcb.4.5.956. [DOI] [PMC free article] [PubMed] [Google Scholar]
  20. Fyrberg C. C., Labeit S., Bullard B., Leonard K., Fyrberg E. Drosophila projectin: relatedness to titin and twitchin and correlation with lethal(4) 102 CDa and bent-dominant mutants. Proc Biol Sci. 1992 Jul 22;249(1324):33–40. doi: 10.1098/rspb.1992.0080. [DOI] [PubMed] [Google Scholar]
  21. Fyrberg E. A., Mahaffey J. W., Bond B. J., Davidson N. Transcripts of the six Drosophila actin genes accumulate in a stage- and tissue-specific manner. Cell. 1983 May;33(1):115–123. doi: 10.1016/0092-8674(83)90340-9. [DOI] [PubMed] [Google Scholar]
  22. Fyrberg E., Fyrberg C. C., Beall C., Saville D. L. Drosophila melanogaster troponin-T mutations engender three distinct syndromes of myofibrillar abnormalities. J Mol Biol. 1990 Dec 5;216(3):657–675. doi: 10.1016/0022-2836(90)90390-8. [DOI] [PubMed] [Google Scholar]
  23. Fyrberg E., Kelly M., Ball E., Fyrberg C., Reedy M. C. Molecular genetics of Drosophila alpha-actinin: mutant alleles disrupt Z disc integrity and muscle insertions. J Cell Biol. 1990 Jun;110(6):1999–2011. doi: 10.1083/jcb.110.6.1999. [DOI] [PMC free article] [PubMed] [Google Scholar]
  24. George E. L., Ober M. B., Emerson C. P., Jr Functional domains of the Drosophila melanogaster muscle myosin heavy-chain gene are encoded by alternatively spliced exons. Mol Cell Biol. 1989 Jul;9(7):2957–2974. doi: 10.1128/mcb.9.7.2957. [DOI] [PMC free article] [PubMed] [Google Scholar]
  25. Hanke P. D., Storti R. V. The Drosophila melanogaster tropomyosin II gene produces multiple proteins by use of alternative tissue-specific promoters and alternative splicing. Mol Cell Biol. 1988 Sep;8(9):3591–3602. doi: 10.1128/mcb.8.9.3591. [DOI] [PMC free article] [PubMed] [Google Scholar]
  26. Hiromi Y., Okamoto H., Gehring W. J., Hotta Y. Germline transformation with Drosophila mutant actin genes induces constitutive expression of heat shock genes. Cell. 1986 Jan 31;44(2):293–301. doi: 10.1016/0092-8674(86)90763-4. [DOI] [PubMed] [Google Scholar]
  27. 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]
  28. Ising G., Block K. Derivation-dependent distribution of insertion sites for a Drosophila transposon. Cold Spring Harb Symp Quant Biol. 1981;45(Pt 2):527–544. doi: 10.1101/sqb.1981.045.01.069. [DOI] [PubMed] [Google Scholar]
  29. Karlik C. C., Coutu M. D., Fyrberg E. A. A nonsense mutation within the act88F actin gene disrupts myofibril formation in Drosophila indirect flight muscles. Cell. 1984 Oct;38(3):711–719. doi: 10.1016/0092-8674(84)90266-6. [DOI] [PubMed] [Google Scholar]
  30. 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]
  31. Karlik C. C., Saville D. L., Fyrberg E. A. Two missense alleles of the Drosophila melanogaster act88F actin gene are strongly antimorphic but only weakly induce synthesis of heat shock proteins. Mol Cell Biol. 1987 Sep;7(9):3084–3091. doi: 10.1128/mcb.7.9.3084. [DOI] [PMC free article] [PubMed] [Google Scholar]
  32. Kronert W. A., Edwards K. A., Roche E. S., Wells L., Bernstein S. I. Muscle-specific accumulation of Drosophila myosin heavy chains: a splicing mutation in an alternative exon results in an isoform substitution. EMBO J. 1991 Sep;10(9):2479–2488. doi: 10.1002/j.1460-2075.1991.tb07787.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  33. 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]
  34. Lindsley D. L., Sandler L., Baker B. S., Carpenter A. T., Denell R. E., Hall J. C., Jacobs P. A., Miklos G. L., Davis B. K., Gethmann R. C. Segmental aneuploidy and the genetic gross structure of the Drosophila genome. Genetics. 1972 May;71(1):157–184. doi: 10.1093/genetics/71.1.157. [DOI] [PMC free article] [PubMed] [Google Scholar]
  35. Mahaffey J. W., Coutu M. D., Fyrberg E. A., Inwood W. The flightless Drosophila mutant raised has two distinct genetic lesions affecting accumulation of myofibrillar proteins in flight muscles. Cell. 1985 Jan;40(1):101–110. doi: 10.1016/0092-8674(85)90313-7. [DOI] [PubMed] [Google Scholar]
  36. Mogami K., Hotta Y. Isolation of Drosophila flightless mutants which affect myofibrillar proteins of indirect flight muscle. Mol Gen Genet. 1981;183(3):409–417. doi: 10.1007/BF00268758. [DOI] [PubMed] [Google Scholar]
  37. Mogami K., O'Donnell P. T., Bernstein S. I., Wright T. R., Emerson C. P., Jr Mutations of the Drosophila myosin heavy-chain gene: effects on transcription, myosin accumulation, and muscle function. Proc Natl Acad Sci U S A. 1986 Mar;83(5):1393–1397. doi: 10.1073/pnas.83.5.1393. [DOI] [PMC free article] [PubMed] [Google Scholar]
  38. Newman R., Butcher G. W., Bullard B., Leonard K. R. A method for determining the periodicity of a troponin component in isolated insect flight muscle thin filaments by gold/Fab labelling. J Cell Sci. 1992 Mar;101(Pt 3):503–508. doi: 10.1242/jcs.101.3.503. [DOI] [PubMed] [Google Scholar]
  39. O'Donnell P. T., Bernstein S. I. Molecular and ultrastructural defects in a Drosophila myosin heavy chain mutant: differential effects on muscle function produced by similar thick filament abnormalities. J Cell Biol. 1988 Dec;107(6 Pt 2):2601–2612. doi: 10.1083/jcb.107.6.2601. [DOI] [PMC free article] [PubMed] [Google Scholar]
  40. O'Donnell P. T., Collier V. L., Mogami K., Bernstein S. I. Ultrastructural and molecular analyses of homozygous-viable Drosophila melanogaster muscle mutants indicate there is a complex pattern of myosin heavy-chain isoform distribution. Genes Dev. 1989 Aug;3(8):1233–1246. doi: 10.1101/gad.3.8.1233. [DOI] [PubMed] [Google Scholar]
  41. Parker V. P., Falkenthal S., Davidson N. Characterization of the myosin light-chain-2 gene of Drosophila melanogaster. Mol Cell Biol. 1985 Nov;5(11):3058–3068. doi: 10.1128/mcb.5.11.3058. [DOI] [PMC free article] [PubMed] [Google Scholar]
  42. Peckham M., Molloy J. E., Sparrow J. C., White D. C. Physiological properties of the dorsal longitudinal flight muscle and the tergal depressor of the trochanter muscle of Drosophila melanogaster. J Muscle Res Cell Motil. 1990 Jun;11(3):203–215. doi: 10.1007/BF01843574. [DOI] [PubMed] [Google Scholar]
  43. Pesacreta T. C., Byers T. J., Dubreuil R., Kiehart D. P., Branton D. Drosophila spectrin: the membrane skeleton during embryogenesis. J Cell Biol. 1989 May;108(5):1697–1709. doi: 10.1083/jcb.108.5.1697. [DOI] [PMC free article] [PubMed] [Google Scholar]
  44. Rozek C. E., Davidson N. Differential processing of RNA transcribed from the single-copy Drosophila myosin heavy chain gene produces four mRNAs that encode two polypeptides. Proc Natl Acad Sci U S A. 1986 Apr;83(7):2128–2132. doi: 10.1073/pnas.83.7.2128. [DOI] [PMC free article] [PubMed] [Google Scholar]
  45. Rozek C. E., Davidson N. Drosophila has one myosin heavy-chain gene with three developmentally regulated transcripts. Cell. 1983 Jan;32(1):23–34. doi: 10.1016/0092-8674(83)90493-2. [DOI] [PubMed] [Google Scholar]
  46. Saide J. D., Chin-Bow S., Hogan-Sheldon J., Busquets-Turner L., Vigoreaux J. O., Valgeirsdottir K., Pardue M. L. Characterization of components of Z-bands in the fibrillar flight muscle of Drosophila melanogaster. J Cell Biol. 1989 Nov;109(5):2157–2167. doi: 10.1083/jcb.109.5.2157. [DOI] [PMC free article] [PubMed] [Google Scholar]
  47. Simpson P. Maternal-Zygotic Gene Interactions during Formation of the Dorsoventral Pattern in Drosophila Embryos. Genetics. 1983 Nov;105(3):615–632. doi: 10.1093/genetics/105.3.615. [DOI] [PMC free article] [PubMed] [Google Scholar]
  48. Sinclair D. A., Suzuki D. T., Grigliatti T. A. Genetic and developmental analysis of a temperature-sensitive minute mutation of Drosophila melanogaster. Genetics. 1981 Mar-Apr;97(3-4):581–606. doi: 10.1093/genetics/97.3-4.581. [DOI] [PMC free article] [PubMed] [Google Scholar]
  49. Sliter T. J., Henrich V. C., Tucker R. L., Gilbert L. I. The genetics of the Dras3-Roughened-ecdysoneless chromosomal region (62B3-4 to 62D3-4) in Drosophila melanogaster: analysis of recessive lethal mutations. Genetics. 1989 Oct;123(2):327–336. doi: 10.1093/genetics/123.2.327. [DOI] [PMC free article] [PubMed] [Google Scholar]
  50. Sparrow J. C., Drummond D. R., Hennessey E. S., Clayton J. D., Lindegaard F. B. Drosophila actin mutants and the study of myofibrillar assembly and function. Symp Soc Exp Biol. 1992;46:111–129. [PubMed] [Google Scholar]
  51. Sparrow J., Reedy M., Ball E., Kyrtatas V., Molloy J., Durston J., Hennessey E., White D. Functional and ultrastructural effects of a missense mutation in the indirect flight muscle-specific actin gene of Drosophila melanogaster. J Mol Biol. 1991 Dec 20;222(4):963–982. doi: 10.1016/0022-2836(91)90588-w. [DOI] [PubMed] [Google Scholar]
  52. Storti R. V., Szwast A. E. Molecular cloning and characterization of Drosophila genes and their expression during embryonic development and in primary muscle cell cultures. Dev Biol. 1982 Apr;90(2):272–283. doi: 10.1016/0012-1606(82)90376-1. [DOI] [PubMed] [Google Scholar]
  53. Toffenetti J., Mischke D., Pardue M. L. Isolation and characterization of the gene for myosin light chain two of Drosophila melanogaster. J Cell Biol. 1987 Jan;104(1):19–28. doi: 10.1083/jcb.104.1.19. [DOI] [PMC free article] [PubMed] [Google Scholar]
  54. Vigoreaux J. O., Saide J. D., Pardue M. L. Structurally different Drosophila striated muscles utilize distinct variants of Z-band-associated proteins. J Muscle Res Cell Motil. 1991 Aug;12(4):340–354. doi: 10.1007/BF01738589. [DOI] [PubMed] [Google Scholar]
  55. Vinós J., Domingo A., Marco R., Cervera M. Identification and characterization of Drosophila melanogaster paramyosin. J Mol Biol. 1991 Aug 5;220(3):687–700. doi: 10.1016/0022-2836(91)90110-r. [DOI] [PubMed] [Google Scholar]
  56. Warmke J. W., Kreuz A. J., Falkenthal S. Co-localization to chromosome bands 99E1-3 of the Drosophila melanogaster myosin light chain-2 gene and a haplo-insufficient locus that affects flight behavior. Genetics. 1989 May;122(1):139–151. doi: 10.1093/genetics/122.1.139. [DOI] [PMC free article] [PubMed] [Google Scholar]
  57. Warmke J., Yamakawa M., Molloy J., Falkenthal S., Maughan D. Myosin light chain-2 mutation affects flight, wing beat frequency, and indirect flight muscle contraction kinetics in Drosophila. J Cell Biol. 1992 Dec;119(6):1523–1539. doi: 10.1083/jcb.119.6.1523. [DOI] [PMC free article] [PubMed] [Google Scholar]
  58. Wassenberg D. R., 2nd, Kronert W. A., O'Donnell P. T., Bernstein S. I. Analysis of the 5' end of the Drosophila muscle myosin heavy chain gene. Alternatively spliced transcripts initiate at a single site and intron locations are conserved compared to myosin genes of other organisms. J Biol Chem. 1987 Aug 5;262(22):10741–10747. [PubMed] [Google Scholar]

Articles from Genetics are provided here courtesy of Oxford University Press

RESOURCES