Summary
Twenty transformed lines have been isolated as a result of the germ line insertion of a 3.2 kb alcohol dehydrogenase (Adh) gene fragment into an Adh negative strain of Drosophila melanogaster by P element-mediated transformation. More than half of these lines exhibited abnormal ADH expression. The level of ADH expression ranges from zero in some lines to near normal levels in others, and the pattern of ADH expression in the larval gut is also abnormal in many of these lines. Each of the abnormal tissue-specific patterns is stable and characterized by the absence or reduction of ADH expression in certain tissues. High levels of ectopic expression were not observed. In two of these lines, the pattern of ADH staining is highly restricted: it is limited to the medial midgut in line MM-50, and to the gastric caecae and the proventriculus in line GC-1. In heterozygotes between these two lines ADH is expressed in both of these tissues. To test the hypothesis that this abnormal expression is due to position effects, inserts were mobilized to new locations. The mobilized inserts exhibited new patterns of tissue-specific expression associated with new cytological insert locations, showing that the abnormal expression in lines MM-50 and GC-1 is due to tissue-specific position effects and not to mutations. The results are discussed in the context of chromatin structure as a possible cause of these position effects.
Similar content being viewed by others
References
Bellen H, O'Kane C, Wilson C, Grossniklaus U, Pearson R, Gehring W (1989) P-element-mediated enhancer detection: a versatile method to study development in Drosophila. Genes Dev 3:1288–1300
Benyajati C, Worcel A (1976) Isolation, characterization and structure of the folded interphase genome of Drosophila melanogaster. Cell 9:393–407
Benyajati C, Place AR, Sofer W (1983) Formaldehyde mutagenesis in Drosophila: Molecular analysis of ADH-negative mutants. Mutat Res 111:1–7
Bier E, Vaessin H, Shepherd S, Lee K, McCall K, Barbel S, Ackerman L, Carretto R, Uemura T, Grell E, Jan L, Jan Y (1989) Searching for pattern and mutation in the Drosophila genome with a P-lacZ vector. Genes Dev 3:1273–1287
Boto L, Cano A, Pestana A (1987) Biochemical and morphological effects of sodium butyrate on Dictyostelium discoidium development. Mol Cell Biochem 74:137–147
Clark SH, Chovnick A (1986) Studies of normal and positionaffected expression of rosy region genes in Drosophila melanogaster. Genetics 114:819–840
Engels W, Preston C, Thompson P, Eggleston W (1986) In situ hybridization to Drosophila salivary chromosomes with biotinylated DNA probes and alkaline phosphatase. Focus (Bethesda Research Laboratories) 8:6–8
Gehring W, Klemenz R, Weber U, Kloter U (1984) Functional analysis of the white + gene of Drosophila by P-factor mediated transformation. EMBO J 3:2077–2085
Goldberg DA (1980) Isolation and partial characterization of the Drosophila alcohol dehydrogenase gene. Proc Natl Acad Sci USA 77:5794–5798
Goldberg DA, Posakony JW, Maniatis T (1983) Correct developmental expression of a cloned alcohol dehydrogenase gene transduced into the Drosophila germ line. Cell 34:59–73
Goldman MA (1988) The chromatin domain as a unit of gene regulation. Bioessays 9:50–55
Hazelrigg T, Levis R, Rubin GM (1984) Transformation of white locus DNA in Drosophila: dosage compensation, interaction with zeste and position effects. Cell 36:469–481
Hofmann JF-X, Laroche T, Brand AH, Gasser SM (1989) RAP-1 factor is necessary for DNA loop formation in vitro at the silent mating type locus HML. Cell 57:725–737
Kellum R, Schedl P (1991) A position-effect assay for boundaries of higher order chromosomal domains. Cell 64:941–950
Krumm A, Roth GE, Korge G (1985) Transformation of salivary gland secretion protein gene Sgs-4 in Drosophila: Stage- and tissue-specific regulation, dosage compensation and position effect. Proc Nat Acad Sci 82:5055–5059
Lester SC, Korn NF, DeMars R (1982) Derepression of genes on the human inactive X chromosome: evidence for differences in locus-specific rates of derepression and rates of transfer of active and inactive genes after DNA-mediated transformation. Somatic Cell Genet 8:265–284
Levis R, Hazelrigg T, Rubin GM (1985) Effects of genomic position on the expression of transduced copies of the white gene of Drosophila. Science 229:558–561
Maniatis T (1982) Molecular cloning: a laboratory Manual. Cold Spring Harbor, Laboratory Press, Cold Spring Harbor, New York
Martin P, Martin A, Osmani A, Sofer W (1986) A transient expression assay for tissue-specific gene expression of alcohol dehydrogenase in Drosophila. Dev Biol 117: 574–580
Martin PF, Martin AR, Sofer W (1985) Abnormal patterns of alcohol dehydrogenase expression in transformed Drosophila. Genetics 110: s24
McNabb SL, Beckendorf SK (1986) Cis-acting sequences which regulate expression of the Sgs-4 glue protein gene of Drosophila. EMBO J 5:2331–2340
Michailidis J, Graves J, Murray ND (1989) Suppression of position-effect variegation in Drosophila melanogaster by fatty acids and dimethylsulphoxide: implications for the mechanism of position effect variegation. J Genet 68:1–8
Mirkovitch J, Mirault M-E, Laemmli UK (1984) Organization of higher order chromatin loop: specific DNA attachment sites on nuclear scaffolds. Cell 39:223–232
Mottus R, Reeves R, Griliatti TA (1980) Butyrate suppression of position effect variegation in Drosophila melanogaster. Mol Gen Genet 178:465–469
O'Hare K, Rubin GM (1983) Structure of P transposable elements and their sites of insertion and excision in the Drosophila melanogaster genome. Cell 34:25–35
O'Kane C, Gehring W (1987) Detection in situ of genomic regulatory elements in Drosophila. Proc Natl Acad Sci 84:9123–9127
Palmiter RD, Brinster RL (1986) Germ line transformation of mice. Annu Rev Genet 20:465–499
Posakony JW, Fisher JA, Maniatis T (1986) Identification of DNA sequences required for the regulation of Drosophila alcohol dehydrogenase gene expression. Cold Spring Harbor Symp Quant Biol 50:515–520
Rubin GM, Spradling AC (1982) Genetic transformation of Drosophila with transposable element vectors. Science 218:348–353
Scholnick SB, Morgan BA, Hirsh J (1983) The cloned dopa decarboxylase gene is developmentally regulated when reintegrated into the Drosophila genome. Cell 34:37–45
Sofer W, Martin PF (1987) Analysis of densitometric data obtained from electrophoretic analysis. Computer Appl Biosci 3:129
Sofer W, Ursprung H (1968) Drosophila alcohol dehydrogenase: purification and partial characterization. J Biol Chem 243:3110–3115
Spradling AC, Rubin GM (1983) The effect of chromosomal position on the expression of the Drosophila xanthine dehydrogenase gene. Cell 34:47–57
Wilson C, Pearson R, Bellen H, O'Kane C, Grossniklaus U, Gehring W (1989) P-element-mediated enhancer detection: an efficient method for isolating and characterizing developmentally regulated genes in Drosophila. Genes Dev 3:1301–1313
Wu C-Y, Mote J, Brennan MD (1990) Tissue-specific expression phenotypes of Hawaiian Drosophila Adh genes in Drosophila melanogaster transformants. Genetics 125:599–610
Author information
Authors and Affiliations
Rights and permissions
About this article
Cite this article
Kirkpatrick, R.B., Martin, P.F. Tissue-specific position effects on alcohol dehydrogenase expression in Drosophila melanogaster . Molec. Gen. Genet. 232, 135–144 (1992). https://doi.org/10.1007/BF00299146
Received:
Issue Date:
DOI: https://doi.org/10.1007/BF00299146