Abstract
The alcohol dehydrogenase (ADH) locus (Adh) of Drosophila melanogaster in polymorphic on a world-wide basis for two allozymes, Fast and Slow. This study was undertaken to determine whether the well-established difference in ADH protein concentration between the allozymes is due to a difference in mRNA levels. RNA gel blot hybridization and an RNase protection assay were used to quantify ADH mRNA levels. Each method used an Adh null mutant as an internal standard. Several Slow and Fast allele pairs of different geographic origins were analyzed. The results provide strong evidence that the ADH protein concentration difference is not accounted for by RNA level.
Full text
PDFImages in this article
Selected References
These references are in PubMed. This may not be the complete list of references from this article.
- Anderson S. M., McDonald J. F. Biochemical and molecular analysis of naturally occurring Adh variants in Drosophila melanogaster. Proc Natl Acad Sci U S A. 1983 Aug;80(15):4798–4802. doi: 10.1073/pnas.80.15.4798. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Aquadro C. F., Desse S. F., Bland M. M., Langley C. H., Laurie-Ahlberg C. C. Molecular population genetics of the alcohol dehydrogenase gene region of Drosophila melanogaster. Genetics. 1986 Dec;114(4):1165–1190. doi: 10.1093/genetics/114.4.1165. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Benyajati C., Place A. R., Sofer W. Formaldehyde mutagenesis in Drosophila. Molecular analysis of ADH-negative mutants. Mutat Res. 1983 Sep;111(1):1–7. doi: 10.1016/0027-5107(83)90002-7. [DOI] [PubMed] [Google Scholar]
- Benyajati C., Place A. R., Wang N., Pentz E., Sofer W. Deletions at intervening sequence splice sites in the alcohol dehydrogenase gene of Drosophila. Nucleic Acids Res. 1982 Nov 25;10(22):7261–7272. doi: 10.1093/nar/10.22.7261. [DOI] [PMC free article] [PubMed] [Google Scholar]
- 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]
- Birley A. J., Couch P. A., Marson A. Genetical variation for enzyme activity in a population of Drosophila melanogaster. VI. Molecular variation in the control of alcohol dehydrogenase (ADH) activity. Heredity (Edinb) 1981 Oct;47(2):185–196. doi: 10.1038/hdy.1981.75. [DOI] [PubMed] [Google Scholar]
- Chia W., Savakis C., Karp R., Pelham H., Ashburner M. Mutation of the Adh gene of Drosophila melanogaster containing an internal tandem duplication. J Mol Biol. 1985 Dec 20;186(4):679–688. doi: 10.1016/0022-2836(85)90388-2. [DOI] [PubMed] [Google Scholar]
- Day T. H., Hillier P. C., Clarke B. The relative quantities and catalytic activities of enzymes produced by alleles at the alcohol dehydrogenase locus in Drosophila melanogaster. Biochem Genet. 1974 Feb;11(2):155–165. doi: 10.1007/BF00485771. [DOI] [PubMed] [Google Scholar]
- Fischer J. A., Maniatis T. Structure and transcription of the Drosophila mulleri alcohol dehydrogenase genes. Nucleic Acids Res. 1985 Oct 11;13(19):6899–6917. doi: 10.1093/nar/13.19.6899. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Fletcher T. S., Ayala F. J., Thatcher D. R., Chambers G. K. Structural analysis of the ADHS electromorph of Drosophila melanogaster. Proc Natl Acad Sci U S A. 1978 Nov;75(11):5609–5612. doi: 10.1073/pnas.75.11.5609. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Gibson J. B. Differences in the number of molecules produced by two allelic electrophoretic enzyme variants in D. melanogaster. Experientia. 1972 Aug 15;28(8):975–976. doi: 10.1007/BF01924982. [DOI] [PubMed] [Google Scholar]
- Goldberg D. A., Posakony J. W., Maniatis T. Correct developmental expression of a cloned alcohol dehydrogenase gene transduced into the Drosophila germ line. Cell. 1983 Aug;34(1):59–73. doi: 10.1016/0092-8674(83)90136-8. [DOI] [PubMed] [Google Scholar]
- Kelley M. R., Mims I. P., Farnet C. M., Dicharry S. A., Lee W. R. Molecular analysis of X-ray-induced alcohol dehydrogenase (ADH) null mutations in Drosophila melanogaster. Genetics. 1985 Feb;109(2):365–377. doi: 10.1093/genetics/109.2.365. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Kreitman M. Nucleotide polymorphism at the alcohol dehydrogenase locus of Drosophila melanogaster. Nature. 1983 Aug 4;304(5925):412–417. doi: 10.1038/304412a0. [DOI] [PubMed] [Google Scholar]
- Laurie-Ahlberg C. C., Maroni G., Bewley G. C., Lucchesi J. C., Weir B. S. Quantitative genetic variation of enzyme activities in natural populations of Drosophila melanogaster. Proc Natl Acad Sci U S A. 1980 Feb;77(2):1073–1077. doi: 10.1073/pnas.77.2.1073. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Laurie-Ahlberg C. C., Stam L. F. Use of P-element-mediated transformation to identify the molecular basis of naturally occurring variants affecting Adh expression in Drosophila melanogaster. Genetics. 1987 Jan;115(1):129–140. doi: 10.1093/genetics/115.1.129. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Lewis N., Gibson J. Variation in amount of enzyme protein in natural populations. Biochem Genet. 1978 Apr;16(3-4):159–170. doi: 10.1007/BF00484075. [DOI] [PubMed] [Google Scholar]
- Mancini G., Carbonara A. O., Heremans J. F. Immunochemical quantitation of antigens by single radial immunodiffusion. Immunochemistry. 1965 Sep;2(3):235–254. doi: 10.1016/0019-2791(65)90004-2. [DOI] [PubMed] [Google Scholar]
- Maroni G. Genetic control of alcohol dehydrogenase levels in Drosophila. Biochem Genet. 1978 Jun;16(5-6):509–523. doi: 10.1007/BF00484215. [DOI] [PubMed] [Google Scholar]
- Maroni G., Laurie-Ahlberg C. C., Adams D. A., Wilton A. N. Genetic variation in the expression of ADH in Drosophila melanogaster. Genetics. 1982 Jul-Aug;101(3-4):431–446. doi: 10.1093/genetics/101.3-4.431. [DOI] [PMC free article] [PubMed] [Google Scholar]
- McDonald J. F., Anderson S. M., Santos M. Biochemical differences between products of the Adh locus in Drosophila. Genetics. 1980 Aug;95(4):1013–1022. doi: 10.1093/genetics/95.4.1013. [DOI] [PMC free article] [PubMed] [Google Scholar]
- McDonald J. F., Ayala F. J. Genetic and biochemical basis of enzyme activity variation in natural populations. I. Alcohol dehydrogenase in Drosophila melanogaster. Genetics. 1978 Jun;89(2):371–388. doi: 10.1093/genetics/89.2.371. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Melton D. A., Krieg P. A., Rebagliati M. R., Maniatis T., Zinn K., Green M. R. Efficient in vitro synthesis of biologically active RNA and RNA hybridization probes from plasmids containing a bacteriophage SP6 promoter. Nucleic Acids Res. 1984 Sep 25;12(18):7035–7056. doi: 10.1093/nar/12.18.7035. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Myers R. M., Larin Z., Maniatis T. Detection of single base substitutions by ribonuclease cleavage at mismatches in RNA:DNA duplexes. Science. 1985 Dec 13;230(4731):1242–1246. doi: 10.1126/science.4071043. [DOI] [PubMed] [Google Scholar]
- Winberg J. O., Hovik R., McKinley-McKee J. S. The alcohol dehydrogenase alleloenzymes AdhS and AdhF from the fruitfly Drosophila melanogaster: an enzymatic rate assay to determine the active-site concentration. Biochem Genet. 1985 Apr;23(3-4):205–216. doi: 10.1007/BF00504319. [DOI] [PubMed] [Google Scholar]