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. 1985 Sep;4(9):2275–2280. doi: 10.1002/j.1460-2075.1985.tb03926.x

Developmentally regulated alternate modes of expression of the Gpdh locus of Drosophila.

G R Skuse, D T Sullivan
PMCID: PMC554497  PMID: 3935431

Abstract

Immunoblot analyses have been performed on extracts prepared from Drosophila melanogaster. Those analyses have revealed two subunit forms of enzyme glycerol 3-phosphate dehydrogenase (GPDH) in larval tissues and in adult abdominal tissues. Thoracic tissue, which accounts for the bulk of the adult GPDH, has only one subunit form, the smaller. The two subunit forms differ by approximately 2400 daltons. In agreement with previous genetic and biochemical data indicating that this enzyme is encoded by a single structural gene, analyses of extracts prepared from a strain carrying a GPDH null mutation detect no GPDH polypeptides in larvae or adults. Similarly, analyses of extracts prepared from a strain carrying a mutation which produces a GPDH polypeptide that differs in size from wild-type reveal a change in the adult thoracic GPDH polypeptide as well as a change in both GPDH polypeptides found in larvae. Total Drosophila RNA prepared from larvae or newly eclosed adults has been translated in a mRNA-dependent cell-free system. GDPH was immunoprecipitated from the translation products and analyzed. Two subunit forms of GPDH were immunoprecipitated from translation products whose synthesis was directed by larval RNA and only one was detected in the polypeptides synthesized from adult RNA. The GPDH polypeptides synthesized in vitro are approximately the same size as the corresponding polypeptides found in vivo. The relative proportion of total GPDH represented by each subunit form synthesized in vitro is similar to those found in vivo.(ABSTRACT TRUNCATED AT 250 WORDS)

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Selected References

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

  1. Benyajati C., Spoerel N., Haymerle H., Ashburner M. The messenger RNA for alcohol dehydrogenase in Drosophila melanogaster differs in its 5' end in different developmental stages. Cell. 1983 May;33(1):125–133. doi: 10.1016/0092-8674(83)90341-0. [DOI] [PubMed] [Google Scholar]
  2. Bewley G. C., Lucchesi J. C. Origin of alpha-glycerophosphate dehydrogenase isozymes in Drosophila melanogaster and their functional relationship in the alpha-glycerophosphate cycle. Biochem Genet. 1977 Apr;15(3-4):235–251. doi: 10.1007/BF00484456. [DOI] [PubMed] [Google Scholar]
  3. Burnette W. N. "Western blotting": electrophoretic transfer of proteins from sodium dodecyl sulfate--polyacrylamide gels to unmodified nitrocellulose and radiographic detection with antibody and radioiodinated protein A. Anal Biochem. 1981 Apr;112(2):195–203. doi: 10.1016/0003-2697(81)90281-5. [DOI] [PubMed] [Google Scholar]
  4. Carlson M., Botstein D. Two differentially regulated mRNAs with different 5' ends encode secreted with intracellular forms of yeast invertase. Cell. 1982 Jan;28(1):145–154. doi: 10.1016/0092-8674(82)90384-1. [DOI] [PubMed] [Google Scholar]
  5. Jellinghaus U., Schätzle U., Schmid W., Roewekamp W. Transcription of a dictyostelium discoidin-i gene in yeast alternative promoter sites used in two different eukaryotic cells. J Mol Biol. 1982 Aug 25;159(4):623–636. doi: 10.1016/0022-2836(82)90104-8. [DOI] [PubMed] [Google Scholar]
  6. Kotarski M. A., Pickert S., Leonard D. A., LaRosa G. J., MacIntyre R. J. The characterization of alpha-glycerophosphate dehydrogenase mutants in Drosophila melanogaster. Genetics. 1983 Oct;105(2):387–407. doi: 10.1093/genetics/105.2.387. [DOI] [PMC free article] [PubMed] [Google Scholar]
  7. Kotarski M. A., Pickert S., MacIntyre R. J. A cytogenetic analysis of the chromosomal region surrounding the alpha-glycerophosphate dehydrogenase locus of Drosophila melanogaster. Genetics. 1983 Oct;105(2):371–386. doi: 10.1093/genetics/105.2.371. [DOI] [PMC free article] [PubMed] [Google Scholar]
  8. 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]
  9. Lai E. C., Roop D. R., Tsai M. J., Woo S. L., O'Malley B. W. Heterogeneous initiation regions for transcription of the chicken ovomucoid gene. Nucleic Acids Res. 1982 Sep 25;10(18):5553–5567. doi: 10.1093/nar/10.18.5553. [DOI] [PMC free article] [PubMed] [Google Scholar]
  10. Laskey R. A., Mills A. D. Quantitative film detection of 3H and 14C in polyacrylamide gels by fluorography. Eur J Biochem. 1975 Aug 15;56(2):335–341. doi: 10.1111/j.1432-1033.1975.tb02238.x. [DOI] [PubMed] [Google Scholar]
  11. Niesel D. W., Bewley G. C., Miller S. G., Armstrong F. B., Lee C. Y. Purification and structural analysis of the soluble sn-glycerol-3-phosphate dehydrogenase isozymes in Drosophila melanogaster. J Biol Chem. 1980 May 10;255(9):4073–4080. [PubMed] [Google Scholar]
  12. Niesel D. W., Pan Y. C., Bewley G. C., Armstrong F. B., Li S. S. Structural analysis of adult and larval isozymes of sn-glycerol-3-phosphate dehydrogenase of Drosophila melanogaster. J Biol Chem. 1982 Jan 25;257(2):979–983. [PubMed] [Google Scholar]
  13. O'Brien S. J., MacIntyre R. J. The -glycerophosphate cycle in Drosophila melanogaster. I. Biochemical and developmental aspects. Biochem Genet. 1972 Oct;7(2):141–161. doi: 10.1007/BF00486085. [DOI] [PubMed] [Google Scholar]
  14. O'Brien S. J., Macintyre R. J. The -glycerophosphate in Drosophila melanogaster. II. Genetic aspects. Genetics. 1972 May;71(1):127–138. doi: 10.1093/genetics/71.1.127. [DOI] [PMC free article] [PubMed] [Google Scholar]
  15. Pelham H. R., Jackson R. J. An efficient mRNA-dependent translation system from reticulocyte lysates. Eur J Biochem. 1976 Aug 1;67(1):247–256. doi: 10.1111/j.1432-1033.1976.tb10656.x. [DOI] [PubMed] [Google Scholar]
  16. Perlman D., Halvorson H. O., Cannon L. E. Presecretory and cytoplasmic invertase polypeptides encoded by distinct mRNAs derived from the same structural gene differ by a signal sequence. Proc Natl Acad Sci U S A. 1982 Feb;79(3):781–785. doi: 10.1073/pnas.79.3.781. [DOI] [PMC free article] [PubMed] [Google Scholar]
  17. 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]
  18. Schwarzbauer J. E., Tamkun J. W., Lemischka I. R., Hynes R. O. Three different fibronectin mRNAs arise by alternative splicing within the coding region. Cell. 1983 Dec;35(2 Pt 1):421–431. doi: 10.1016/0092-8674(83)90175-7. [DOI] [PubMed] [Google Scholar]
  19. Sullivan D. T., Donovan F. A., Skuse G. Developmental regulation of glycerol-3-phosphate dehydrogenase synthesis in Drosophila. Biochem Genet. 1983 Feb;21(1-2):49–62. doi: 10.1007/BF02395391. [DOI] [PubMed] [Google Scholar]
  20. Towbin H., Staehelin T., Gordon J. Electrophoretic transfer of proteins from polyacrylamide gels to nitrocellulose sheets: procedure and some applications. Proc Natl Acad Sci U S A. 1979 Sep;76(9):4350–4354. doi: 10.1073/pnas.76.9.4350. [DOI] [PMC free article] [PubMed] [Google Scholar]
  21. Wright D. A., Shaw C. R. Genetics and ontogeny of alpha-glycerophosphate dehydrogenase isozymes in Drosophila melanogaster. Biochem Genet. 1969 Aug;3(4):343–353. doi: 10.1007/BF00485718. [DOI] [PubMed] [Google Scholar]

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