Skip to main content
PMC home page
Molecular and Cellular Biology logoLink to Molecular and Cellular Biology
. 1993 Mar;13(3):1433–1439. doi: 10.1128/mcb.13.3.1433

Abnormal muscle development in the heldup3 mutant of Drosophila melanogaster is caused by a splicing defect affecting selected troponin I isoforms.

J A Barbas 1, J Galceran 1, L Torroja 1, A Prado 1, A Ferrús 1
PMCID: PMC359453  PMID: 7680094

Abstract

The troponin I (TnI) gene of Drosophila melanogaster encodes a family of 10 isoforms resulting from the differential splicing of 13 exons. Four of these exons (6a1, 6a2, 6b1, and 6b2) are mutually exclusive and very similar in sequence. TnI isoforms show qualitative specificity whereby each muscle expresses a selected repertoire of them. In addition, TnI isoforms show quantitative specificity whereby each muscle expresses characteristic amounts of each isoform. In the mutant heldup3, the development of the thoracic muscles DLM, DVM, and TDT is aborted. The mutation consists of a one-nucleotide displacement of the 3' AG splice site at the intron preceding exon 6b1, resulting in the failure to produce all exon 6b1-containing TnI isoforms. These molecular changes in a constituent of the thin filaments cause the selective failure to develop the DLM, DVM, and TDT muscles while having no visible effect on other muscles wherein exon 6b1 expression is minor.

Full text

PDF
1437

Images in this article

1435Image
on p.1435
1436Image
on p.1436
1436Image
on p.1436
1436Image
on p.1436
1437Image
on p.1437

Selected References

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

  1. Andrews L. G., Markert M. L. Exon skipping in purine nucleoside phosphorylase mRNA processing leading to severe immunodeficiency. J Biol Chem. 1992 Apr 15;267(11):7834–7838. [PubMed] [Google Scholar]
  2. 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]
  3. Baumann A., Krah-Jentgens I., Müller R., Müller-Holtkamp F., Seidel R., Kecskemethy N., Casal J., Ferrus A., Pongs O. Molecular organization of the maternal effect region of the Shaker complex of Drosophila: characterization of an I(A) channel transcript with homology to vertebrate Na channel. EMBO J. 1987 Nov;6(11):3419–3429. doi: 10.1002/j.1460-2075.1987.tb02665.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  4. 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]
  5. 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]
  6. Chomczynski P., Sacchi N. Single-step method of RNA isolation by acid guanidinium thiocyanate-phenol-chloroform extraction. Anal Biochem. 1987 Apr;162(1):156–159. doi: 10.1006/abio.1987.9999. [DOI] [PubMed] [Google Scholar]
  7. Cole W. G., Chiodo A. A., Lamande S. R., Janeczko R., Ramirez F., Dahl H. H., Chan D., Bateman J. F. A base substitution at a splice site in the COL3A1 gene causes exon skipping and generates abnormal type III procollagen in a patient with Ehlers-Danlos syndrome type IV. J Biol Chem. 1990 Oct 5;265(28):17070–17077. [PubMed] [Google Scholar]
  8. 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]
  9. Deak I. I. Mutations of Drosophila melanogaster that affect muscles. J Embryol Exp Morphol. 1977 Aug;40:35–63. [PubMed] [Google Scholar]
  10. Feinberg A. P., Vogelstein B. A technique for radiolabeling DNA restriction endonuclease fragments to high specific activity. Anal Biochem. 1983 Jul 1;132(1):6–13. doi: 10.1016/0003-2697(83)90418-9. [DOI] [PubMed] [Google Scholar]
  11. Ferrús A., Llamazares S., de la Pompa J. L., Tanouye M. A., Pongs O. Genetic analysis of the Shaker gene complex of Drosophila melanogaster. Genetics. 1990 Jun;125(2):383–398. doi: 10.1093/genetics/125.2.383. [DOI] [PMC free article] [PubMed] [Google Scholar]
  12. Herring W. J., McKean M., Dracopoli N., Danner D. J. Branched chain acyltransferase absence due to an Alu-based genomic deletion allele and an exon skipping allele in a compound heterozygote proband expressing maple syrup urine disease. Biochim Biophys Acta. 1992 Mar 20;1138(3):236–242. doi: 10.1016/0925-4439(92)90043-m. [DOI] [PubMed] [Google Scholar]
  13. 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]
  14. Horwitz J., Bullard B., Mercola D. Interaction of troponin subunits. The interaction between the inhibitory and tropomyosin-binding subunits. J Biol Chem. 1979 Jan 25;254(2):350–355. [PubMed] [Google Scholar]
  15. Karlik C. C., Fyrberg E. A. Two Drosophila melanogaster tropomyosin genes: structural and functional aspects. Mol Cell Biol. 1986 Jun;6(6):1965–1973. doi: 10.1128/mcb.6.6.1965. [DOI] [PMC free article] [PubMed] [Google Scholar]
  16. Lichtinghagen R., Stocker M., Wittka R., Boheim G., Stühmer W., Ferrus A., Pongs O. Molecular basis of altered excitability in Shaker mutants of Drosophila melanogaster. EMBO J. 1990 Dec;9(13):4399–4407. doi: 10.1002/j.1460-2075.1990.tb07890.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  17. Lim S. K., Maquat L. E. Human beta-globin mRNAs that harbor a nonsense codon are degraded in murine erythroid tissues to intermediates lacking regions of exon I or exons I and II that have a cap-like structure at the 5' termini. EMBO J. 1992 Sep;11(9):3271–3278. doi: 10.1002/j.1460-2075.1992.tb05405.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  18. Nicholls A. C., Oliver J., Renouf D. V., Heath D. A., Pope F. M. The molecular defect in a family with mild atypical osteogenesis imperfecta and extreme joint hypermobility: exon skipping caused by an 11-bp deletion from an intron in one COL1A2 allele. Hum Genet. 1992 Mar;88(6):627–633. doi: 10.1007/BF02265286. [DOI] [PubMed] [Google Scholar]
  19. 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]
  20. Pongs O., Kecskemethy N., Müller R., Krah-Jentgens I., Baumann A., Kiltz H. H., Canal I., Llamazares S., Ferrus A. Shaker encodes a family of putative potassium channel proteins in the nervous system of Drosophila. EMBO J. 1988 Apr;7(4):1087–1096. doi: 10.1002/j.1460-2075.1988.tb02917.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  21. Reed R. The organization of 3' splice-site sequences in mammalian introns. Genes Dev. 1989 Dec;3(12B):2113–2123. doi: 10.1101/gad.3.12b.2113. [DOI] [PubMed] [Google Scholar]
  22. Rüther U., Müller-Hill B. Easy identification of cDNA clones. EMBO J. 1983;2(10):1791–1794. doi: 10.1002/j.1460-2075.1983.tb01659.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  23. Sakuraba H., Eng C. M., Desnick R. J., Bishop D. F. Invariant exon skipping in the human alpha-galactosidase A pre-mRNA: Ag+1 to t substitution in a 5'-splice site causing Fabry disease. Genomics. 1992 Apr;12(4):643–650. doi: 10.1016/0888-7543(92)90288-4. [DOI] [PubMed] [Google Scholar]
  24. 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]
  25. Smith C. W., Porro E. B., Patton J. G., Nadal-Ginard B. Scanning from an independently specified branch point defines the 3' splice site of mammalian introns. Nature. 1989 Nov 16;342(6247):243–247. doi: 10.1038/342243a0. [DOI] [PubMed] [Google Scholar]
  26. Steingrimsdottir H., Rowley G., Dorado G., Cole J., Lehmann A. R. Mutations which alter splicing in the human hypoxanthine-guanine phosphoribosyltransferase gene. Nucleic Acids Res. 1992 Mar 25;20(6):1201–1208. doi: 10.1093/nar/20.6.1201. [DOI] [PMC free article] [PubMed] [Google Scholar]
  27. Zot A. S., Potter J. D. Structural aspects of troponin-tropomyosin regulation of skeletal muscle contraction. Annu Rev Biophys Biophys Chem. 1987;16:535–559. doi: 10.1146/annurev.bb.16.060187.002535. [DOI] [PubMed] [Google Scholar]
  28. de la Pompa J. L., Garcia J. R., Ferrús A. Genetic analysis of muscle development in Drosophila melanogaster. Dev Biol. 1989 Feb;131(2):439–454. doi: 10.1016/s0012-1606(89)80016-8. [DOI] [PubMed] [Google Scholar]

Articles from Molecular and Cellular Biology are provided here courtesy of Taylor & Francis

RESOURCES