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. 1986 Mar;5(3):589–596. doi: 10.1002/j.1460-2075.1986.tb04251.x

Molecular characterization of mutant actin genes which induce heat-shock proteins in Drosophila flight muscles

Hitoshi Okamoto 1, Yasushi Hiromi 1, Etsuko Ishikawa 1, Takuma Yamada 1, Kazuyoshi Isoda 1, Hideaki Maekawa 2, Yoshiki Hotta 1
PMCID: PMC1166803  PMID: 16453675

Abstract

Heat-shock proteins (hsps) are constitutively induced by the mutant actins in the Drosophila indirect flight muscles (IFM). We compared primary structures of the mutant actin genes (KM75 and HH5) which induce hsps and of the non-inducing alleles (KM129 and KM88). The KM75 actin has lost 20 amino acids at the C-terminus. The HH5 actin has only one amino acid substitution, from Gly-336 to Ser. In KM129, the C-terminal part of actin is replaced by novel amino acids. KM88 is a null allele, with an amber mutation early in the coding region of the mutated actin gene. Although all of the KM75, HH5 and KM129 actins have defects near the C-terminus, only hsp-inducing mutant actins cause enlargement of the IFM nuclei as well as a disruption of myofibrils even in the presence of two copies of the normal genes. We further consider the underlying mechanisms linking these features of the hsp-inducing alleles.

Keywords: actin gene, heat shock protein, Drosophila

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Selected References

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

  1. Ashburner M., Bonner J. J. The induction of gene activity in drosophilia by heat shock. Cell. 1979 Jun;17(2):241–254. doi: 10.1016/0092-8674(79)90150-8. [DOI] [PubMed] [Google Scholar]
  2. Baumann B., Potash M. J., Köhler G. Consequences of frameshift mutations at the immunoglobulin heavy chain locus of the mouse. EMBO J. 1985 Feb;4(2):351–359. doi: 10.1002/j.1460-2075.1985.tb03636.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  3. Bienz M., Pelham H. R. Expression of a Drosophila heat-shock protein in Xenopus oocytes: conserved and divergent regulatory signals. EMBO J. 1982;1(12):1583–1588. doi: 10.1002/j.1460-2075.1982.tb01359.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  4. Bonner J. J., Parks C., Parker-Thornburg J., Mortin M. A., Pelham H. R. The use of promoter fusions in Drosophila genetics: isolation of mutations affecting the heat shock response. Cell. 1984 Jul;37(3):979–991. doi: 10.1016/0092-8674(84)90432-x. [DOI] [PubMed] [Google Scholar]
  5. Breathnach R., Chambon P. Organization and expression of eucaryotic split genes coding for proteins. Annu Rev Biochem. 1981;50:349–383. doi: 10.1146/annurev.bi.50.070181.002025. [DOI] [PubMed] [Google Scholar]
  6. Chirgwin J. M., Przybyla A. E., MacDonald R. J., Rutter W. J. Isolation of biologically active ribonucleic acid from sources enriched in ribonuclease. Biochemistry. 1979 Nov 27;18(24):5294–5299. doi: 10.1021/bi00591a005. [DOI] [PubMed] [Google Scholar]
  7. De Robertis E. M., Longthorne R. F., Gurdon J. B. Intracellular migration of nuclear proteins in Xenopus oocytes. Nature. 1978 Mar 16;272(5650):254–256. doi: 10.1038/272254a0. [DOI] [PubMed] [Google Scholar]
  8. Egly J. M., Miyamoto N. G., Moncollin V., Chambon P. Is actin a transcription initiation factor for RNA polymerase B? EMBO J. 1984 Oct;3(10):2363–2371. doi: 10.1002/j.1460-2075.1984.tb02141.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  9. Elzinga M., Phelan J. J. F-actin is intermolecularly crosslinked by N,N'-p-phenylenedimaleimide through lysine-191 and cysteine-374. Proc Natl Acad Sci U S A. 1984 Nov;81(21):6599–6602. doi: 10.1073/pnas.81.21.6599. [DOI] [PMC free article] [PubMed] [Google Scholar]
  10. Frischauf A. M., Garoff H., Lehrach H. A subcloning strategy for DNA sequence analysis. Nucleic Acids Res. 1980 Dec 11;8(23):5541–5549. doi: 10.1093/nar/8.23.5541. [DOI] [PMC free article] [PubMed] [Google Scholar]
  11. Fujita S. C., Zipursky S. L., Benzer S., Ferrús A., Shotwell S. L. Monoclonal antibodies against the Drosophila nervous system. Proc Natl Acad Sci U S A. 1982 Dec;79(24):7929–7933. doi: 10.1073/pnas.79.24.7929. [DOI] [PMC free article] [PubMed] [Google Scholar]
  12. Fyrberg E. A., Bond B. J., Hershey N. D., Mixter K. S., Davidson N. The actin genes of Drosophila: protein coding regions are highly conserved but intron positions are not. Cell. 1981 Apr;24(1):107–116. doi: 10.1016/0092-8674(81)90506-7. [DOI] [PubMed] [Google Scholar]
  13. 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]
  14. Hiromi Y., Hotta Y. Actin gene mutations in Drosophila; heat shock activation in the indirect flight muscles. EMBO J. 1985 Jul;4(7):1681–1687. doi: 10.1002/j.1460-2075.1985.tb03837.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  15. Hotta Y., Benzer S. Mapping of behavior in Drosophila mosaics. Symp Soc Dev Biol. 1973;31:129–167. doi: 10.1016/b978-0-12-612975-5.50010-x. [DOI] [PubMed] [Google Scholar]
  16. 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]
  17. Keller E. B., Noon W. A. Intron splicing: a conserved internal signal in introns of animal pre-mRNAs. Proc Natl Acad Sci U S A. 1984 Dec;81(23):7417–7420. doi: 10.1073/pnas.81.23.7417. [DOI] [PMC free article] [PubMed] [Google Scholar]
  18. Koana T., Hotta Y. Isolation and characterization of flightless mutants in Drosophila melanogaster. J Embryol Exp Morphol. 1978 Jun;45:123–143. [PubMed] [Google Scholar]
  19. Landel C. P., Krause M., Waterston R. H., Hirsh D. DNA rearrangements of the actin gene cluster in Caenorhabditis elegans accompany reversion of three muscle mutants. J Mol Biol. 1984 Dec 15;180(3):497–513. doi: 10.1016/0022-2836(84)90024-x. [DOI] [PubMed] [Google Scholar]
  20. Leavitt J., Gunning P., Porreca P., Ng S. Y., Lin C. S., Kedes L. Molecular cloning and characterization of mutant and wild-type human beta-actin genes. Mol Cell Biol. 1984 Oct;4(10):1961–1969. doi: 10.1128/mcb.4.10.1961. [DOI] [PMC free article] [PubMed] [Google Scholar]
  21. Leavitt J., Kakunaga T. Expression of a variant form of actin and additional polypeptide changes following chemical-induced in vitro neoplastic transformation of human fibroblasts. J Biol Chem. 1980 Feb 25;255(4):1650–1661. [PubMed] [Google Scholar]
  22. Losson R., Lacroute F. Interference of nonsense mutations with eukaryotic messenger RNA stability. Proc Natl Acad Sci U S A. 1979 Oct;76(10):5134–5137. doi: 10.1073/pnas.76.10.5134. [DOI] [PMC free article] [PubMed] [Google Scholar]
  23. 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]
  24. Maruta H., Knoerzer W., Hinssen H., Isenberg G. Regulation of actin polymerization by non-polymerizable actin-like proteins. 1984 Nov 29-Dec 5Nature. 312(5993):424–427. doi: 10.1038/312424a0. [DOI] [PubMed] [Google Scholar]
  25. Maxam A. M., Gilbert W. A new method for sequencing DNA. Proc Natl Acad Sci U S A. 1977 Feb;74(2):560–564. doi: 10.1073/pnas.74.2.560. [DOI] [PMC free article] [PubMed] [Google Scholar]
  26. Messing J. New M13 vectors for cloning. Methods Enzymol. 1983;101:20–78. doi: 10.1016/0076-6879(83)01005-8. [DOI] [PubMed] [Google Scholar]
  27. 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]
  28. Mount S. M. A catalogue of splice junction sequences. Nucleic Acids Res. 1982 Jan 22;10(2):459–472. doi: 10.1093/nar/10.2.459. [DOI] [PMC free article] [PubMed] [Google Scholar]
  29. Nathans J., Hogness D. S. Isolation, sequence analysis, and intron-exon arrangement of the gene encoding bovine rhodopsin. Cell. 1983 Oct;34(3):807–814. doi: 10.1016/0092-8674(83)90537-8. [DOI] [PubMed] [Google Scholar]
  30. Parker C. S., Topol J. A Drosophila RNA polymerase II transcription factor binds to the regulatory site of an hsp 70 gene. Cell. 1984 May;37(1):273–283. doi: 10.1016/0092-8674(84)90323-4. [DOI] [PubMed] [Google Scholar]
  31. Pelham H. R. A regulatory upstream promoter element in the Drosophila hsp 70 heat-shock gene. Cell. 1982 Sep;30(2):517–528. doi: 10.1016/0092-8674(82)90249-5. [DOI] [PubMed] [Google Scholar]
  32. Pelham H. R., Bienz M. A synthetic heat-shock promoter element confers heat-inducibility on the herpes simplex virus thymidine kinase gene. EMBO J. 1982;1(11):1473–1477. doi: 10.1002/j.1460-2075.1982.tb01340.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  33. Rubin R. W., Hill M. C., Hepworth P., Boehmer J. Isolation and electrophoretic analysis of nucleoli, phenol-soluble nuclear proteins, and outer cyst walls from Acanthamoeba castellanii during encystation initiation. J Cell Biol. 1976 Mar;68(3):740–751. doi: 10.1083/jcb.68.3.740. [DOI] [PMC free article] [PubMed] [Google Scholar]
  34. Sanchez F., Tobin S. L., Rdest U., Zulauf E., McCarthy B. J. Two Drosophila actin genes in detail. Gene structure, protein structure and transcription during development. J Mol Biol. 1983 Feb 5;163(4):533–551. doi: 10.1016/0022-2836(83)90111-0. [DOI] [PubMed] [Google Scholar]
  35. Scheer U., Hinssen H., Franke W. W., Jockusch B. M. Microinjection of actin-binding proteins and actin antibodies demonstrates involvement of nuclear actin in transcription of lampbrush chromosomes. Cell. 1984 Nov;39(1):111–122. doi: 10.1016/0092-8674(84)90196-x. [DOI] [PubMed] [Google Scholar]
  36. Southern E. M. Detection of specific sequences among DNA fragments separated by gel electrophoresis. J Mol Biol. 1975 Nov 5;98(3):503–517. doi: 10.1016/s0022-2836(75)80083-0. [DOI] [PubMed] [Google Scholar]
  37. Sutoh K. Actin-actin and actin-deoxyribonuclease I contact sites in the actin sequence. Biochemistry. 1984 Apr 24;23(9):1942–1946. doi: 10.1021/bi00304a009. [DOI] [PubMed] [Google Scholar]
  38. Suzuki Y., Gage L. P., Brown D. D. The genes for silk fibroin in Bombyx mori. J Mol Biol. 1972 Oct 14;70(3):637–649. doi: 10.1016/0022-2836(72)90563-3. [DOI] [PubMed] [Google Scholar]
  39. Thomas P. S. Hybridization of denatured RNA and small DNA fragments transferred to nitrocellulose. Proc Natl Acad Sci U S A. 1980 Sep;77(9):5201–5205. doi: 10.1073/pnas.77.9.5201. [DOI] [PMC free article] [PubMed] [Google Scholar]
  40. Tobin S. L., Zulauf E., Sánchez F., Craig E. A., McCarthy B. J. Multiple actin-related sequences in the Drosophila melanogaster genome. Cell. 1980 Jan;19(1):121–131. doi: 10.1016/0092-8674(80)90393-1. [DOI] [PubMed] [Google Scholar]
  41. Vandekerckhove J., Leavitt J., Kakunaga T., Weber K. Coexpression of a mutant beta-actin and the two normal beta- and gamma-cytoplasmic actins in a stably transformed human cell line. Cell. 1980 Dec;22(3):893–899. doi: 10.1016/0092-8674(80)90566-8. [DOI] [PubMed] [Google Scholar]
  42. Waterston R. H., Hirsh D., Lane T. R. Dominant mutations affecting muscle structure in Caenorhabditis elegans that map near the actin gene cluster. J Mol Biol. 1984 Dec 15;180(3):473–496. doi: 10.1016/0022-2836(84)90023-8. [DOI] [PubMed] [Google Scholar]
  43. Wu C. Two protein-binding sites in chromatin implicated in the activation of heat-shock genes. Nature. 1984 May 17;309(5965):229–234. doi: 10.1038/309229a0. [DOI] [PubMed] [Google Scholar]

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