Abstract
The sex of Drosophila melanogaster is determined by a hierarchy of genes. The ultimate targets of this regulatory hierarchy are the genes encoding terminal differentiation products of one sex. For one of the best-characterized target genes, that encoding female-specific yolk protein 1 (YP1), sex-specific transcriptional controls have been clearly demonstrated. In addition, sex-specific posttranscriptional controls were suggested from experiments in which YP1 RNA was induced in males with hormones. To determine whether males can efficiently process and translate a transcript which is normally found only in females, we used a non-sex-specific promoter, the hsp70 gene promoter, to drive YP1 gene transcription in germ line transformed males. The efficiency of expression of the YP1 gene at levels of RNA splicing, translation, and protein secretion in these males was compared with that in wild-type females. These experiments show that there are no sex-specific posttranscriptional controls operating to limit the production of secreted YP1 in males. Promoters containing different numbers of heat shock elements were tested for their ability to drive YP1 gene transcription in males. These results show that incompatibility between the hsp70 gene heat shock elements and the YP1 gene promoter can be overcome by increasing the amount of hsp70 gene sequence up or downstream of the TATA box. In the course of this study, two vectors useful for placing genes under heat shock regulation were constructed. One of these vectors is designed so that the heat-induced transcript produced is the "authentic" primary transcript; it should be useful for studies of posttranscriptional regulation.
Full text
PDF








Images in this article
Selected References
These references are in PubMed. This may not be the complete list of references from this article.
- Baker B. S., Nagoshi R. N., Burtis K. C. Molecular genetic aspects of sex determination in Drosophila. Bioessays. 1987 Feb;6(2):66–70. doi: 10.1002/bies.950060206. [DOI] [PubMed] [Google Scholar]
- Barnett T., Pachl C., Gergen J. P., Wensink P. C. The isolation and characterization of Drosophila yolk protein genes. Cell. 1980 Oct;21(3):729–738. doi: 10.1016/0092-8674(80)90436-5. [DOI] [PubMed] [Google Scholar]
- Beall C. J., Hirsh J. Regulation of the Drosophila dopa decarboxylase gene in neuronal and glial cells. Genes Dev. 1987 Jul;1(5):510–520. doi: 10.1101/gad.1.5.510. [DOI] [PubMed] [Google Scholar]
- Belote J. M., Handler A. M., Wolfner M. F., Livak K. J., Baker B. S. Sex-specific regulation of yolk protein gene expression in Drosophila. Cell. 1985 Feb;40(2):339–348. doi: 10.1016/0092-8674(85)90148-5. [DOI] [PubMed] [Google Scholar]
- Bienz M., Pelham H. R. Heat shock regulatory elements function as an inducible enhancer in the Xenopus hsp70 gene and when linked to a heterologous promoter. Cell. 1986 Jun 6;45(5):753–760. doi: 10.1016/0092-8674(86)90789-0. [DOI] [PubMed] [Google Scholar]
- Boggs R. T., Gregor P., Idriss S., Belote J. M., McKeown M. Regulation of sexual differentiation in D. melanogaster via alternative splicing of RNA from the transformer gene. Cell. 1987 Aug 28;50(5):739–747. doi: 10.1016/0092-8674(87)90332-1. [DOI] [PubMed] [Google Scholar]
- 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]
- Boswell R. E., Mahowald A. P. tudor, a gene required for assembly of the germ plasm in Drosophila melanogaster. Cell. 1985 Nov;43(1):97–104. doi: 10.1016/0092-8674(85)90015-7. [DOI] [PubMed] [Google Scholar]
- Bownes M., Blair M., Kozma R., Dempster M. 20-hydroxyecdysone stimulates tissue-specific yolk-protein gene transcription in both male and female Drosophila. J Embryol Exp Morphol. 1983 Dec;78:249–268. [PubMed] [Google Scholar]
- Brennan M. D., Weiner A. J., Goralski T. J., Mahowald A. P. The follicle cells are a major site of vitellogenin synthesis in Drosophila melanogaster. Dev Biol. 1982 Jan;89(1):225–236. doi: 10.1016/0012-1606(82)90309-8. [DOI] [PubMed] [Google Scholar]
- Brock M. L., Shapiro D. J. Estrogen stabilizes vitellogenin mRNA against cytoplasmic degradation. Cell. 1983 Aug;34(1):207–214. doi: 10.1016/0092-8674(83)90151-4. [DOI] [PubMed] [Google Scholar]
- Chapman K. B., Wolfner M. F. Determination of male-specific gene expression in Drosophila accessory glands. Dev Biol. 1988 Mar;126(1):195–202. doi: 10.1016/0012-1606(88)90253-9. [DOI] [PubMed] [Google Scholar]
- DiBenedetto A. J., Lakich D. M., Kruger W. D., Belote J. M., Baker B. S., Wolfner M. F. Sequences expressed sex-specifically in Drosophila melanogaster adults. Dev Biol. 1987 Jan;119(1):242–251. doi: 10.1016/0012-1606(87)90225-9. [DOI] [PubMed] [Google Scholar]
- Dudler R., Travers A. A. Upstream elements necessary for optimal function of the hsp 70 promoter in transformed flies. Cell. 1984 Sep;38(2):391–398. doi: 10.1016/0092-8674(84)90494-x. [DOI] [PubMed] [Google Scholar]
- Feinberg A. P., Vogelstein B. "A technique for radiolabeling DNA restriction endonuclease fragments to high specific activity". Addendum. Anal Biochem. 1984 Feb;137(1):266–267. doi: 10.1016/0003-2697(84)90381-6. [DOI] [PubMed] [Google Scholar]
- Garabedian M. J., Hung M. C., Wensink P. C. Independent control elements that determine yolk protein gene expression in alternative Drosophila tissues. Proc Natl Acad Sci U S A. 1985 Mar;82(5):1396–1400. doi: 10.1073/pnas.82.5.1396. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Garabedian M. J., Shepherd B. M., Wensink P. C. A tissue-specific transcription enhancer from the Drosophila yolk protein 1 gene. Cell. 1986 Jun 20;45(6):859–867. doi: 10.1016/0092-8674(86)90560-x. [DOI] [PubMed] [Google Scholar]
- Glaser R. L., Wolfner M. F., Lis J. T. Spatial and temporal pattern of hsp26 expression during normal development. EMBO J. 1986 Apr;5(4):747–754. doi: 10.1002/j.1460-2075.1986.tb04277.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Hackett R. W., Lis J. T. Localization of the hsp83 transcript within a 3292 nucleotide sequence from the 63B heat shock locus of D. melanogaster. Nucleic Acids Res. 1983 Oct 25;11(20):7011–7030. doi: 10.1093/nar/11.20.7011. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Hultmark D., Klemenz R., Gehring W. J. Translational and transcriptional control elements in the untranslated leader of the heat-shock gene hsp22. Cell. 1986 Feb 14;44(3):429–438. doi: 10.1016/0092-8674(86)90464-2. [DOI] [PubMed] [Google Scholar]
- Hung M. C., Wensink P. C. The sequence of the Drosophila melanogaster gene for yolk protein 1. Nucleic Acids Res. 1981 Dec 11;9(23):6407–6419. doi: 10.1093/nar/9.23.6407. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Ish-Horowicz D., Pinchin S. M. Pattern abnormalities induced by ectopic expression of the Drosophila gene hairy are associated with repression of ftz transcription. Cell. 1987 Nov 6;51(3):405–415. doi: 10.1016/0092-8674(87)90636-2. [DOI] [PubMed] [Google Scholar]
- Klemenz R., Hultmark D., Gehring W. J. Selective translation of heat shock mRNA in Drosophila melanogaster depends on sequence information in the leader. EMBO J. 1985 Aug;4(8):2053–2060. doi: 10.1002/j.1460-2075.1985.tb03891.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Legerski R. J., Hodnett J. L., Gray H. B., Jr Extracellular nucleases of pseudomonas BAL 31. III. Use of the double-strand deoxyriboexonuclease activity as the basis of a convenient method for the mapping of fragments of DNA produced by cleavage with restriction enzymes. Nucleic Acids Res. 1978 May;5(5):1445–1464. doi: 10.1093/nar/5.5.1445. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Lis J. T., Neckameyer W., Dubensky R., Costlow N. Cloning and characterization of nine heat-shock-induced mRNAs of Drosophila melanogaster. Gene. 1981 Oct;15(1):67–80. doi: 10.1016/0378-1119(81)90105-0. [DOI] [PubMed] [Google Scholar]
- Lis J. T., Simon J. A., Sutton C. A. New heat shock puffs and beta-galactosidase activity resulting from transformation of Drosophila with an hsp70-lacZ hybrid gene. Cell. 1983 Dec;35(2 Pt 1):403–410. doi: 10.1016/0092-8674(83)90173-3. [DOI] [PubMed] [Google Scholar]
- Mahowald A. P. Ultrastructural observations on oogenesis in Drosophila. J Morphol. 1972 May;137(1):29–48. doi: 10.1002/jmor.1051370103. [DOI] [PubMed] [Google Scholar]
- McGarry T. J., Lindquist S. The preferential translation of Drosophila hsp70 mRNA requires sequences in the untranslated leader. Cell. 1985 Oct;42(3):903–911. doi: 10.1016/0092-8674(85)90286-7. [DOI] [PubMed] [Google Scholar]
- McKnight S. L., Gavis E. R., Kingsbury R., Axel R. Analysis of transcriptional regulatory signals of the HSV thymidine kinase gene: identification of an upstream control region. Cell. 1981 Aug;25(2):385–398. doi: 10.1016/0092-8674(81)90057-x. [DOI] [PubMed] [Google Scholar]
- Monsma S. A., Ard R., Lis J. T., Wolfner M. F. Localized heat-shock induction in Drosophila melanogaster. J Exp Zool. 1988 Sep;247(3):279–284. doi: 10.1002/jez.1402470312. [DOI] [PubMed] [Google Scholar]
- O'Farrell P. H. High resolution two-dimensional electrophoresis of proteins. J Biol Chem. 1975 May 25;250(10):4007–4021. [PMC free article] [PubMed] [Google Scholar]
- 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]
- Postlethwait J. H., Laugé G., Handler A. M. Yolk protein synthesis in ovariectomized and genetically agametic [X87]Drosophila melanogaster. Gen Comp Endocrinol. 1980 Apr;40(4):385–390. doi: 10.1016/0016-6480(80)90001-5. [DOI] [PubMed] [Google Scholar]
- Reddy P., Zehring W. A., Wheeler D. A., Pirrotta V., Hadfield C., Hall J. C., Rosbash M. Molecular analysis of the period locus in Drosophila melanogaster and identification of a transcript involved in biological rhythms. Cell. 1984 Oct;38(3):701–710. doi: 10.1016/0092-8674(84)90265-4. [DOI] [PubMed] [Google Scholar]
- 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]
- Schneuwly S., Klemenz R., Gehring W. J. Redesigning the body plan of Drosophila by ectopic expression of the homoeotic gene Antennapedia. 1987 Feb 26-Mar 4Nature. 325(6107):816–818. doi: 10.1038/325816a0. [DOI] [PubMed] [Google Scholar]
- Seifert H. S., Chen E. Y., So M., Heffron F. Shuttle mutagenesis: a method of transposon mutagenesis for Saccharomyces cerevisiae. Proc Natl Acad Sci U S A. 1986 Feb;83(3):735–739. doi: 10.1073/pnas.83.3.735. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Shirk P. D., Minoo P., Postlethwait J. H. 20-Hydroxyecdysone stimulates the accumulation of translatable yolk polypeptide gene transcript in adult male Drosophila melanogaster. Proc Natl Acad Sci U S A. 1983 Jan;80(1):186–190. doi: 10.1073/pnas.80.1.186. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Simon J. A., Lis J. T. A germline transformation analysis reveals flexibility in the organization of heat shock consensus elements. Nucleic Acids Res. 1987 Apr 10;15(7):2971–2988. doi: 10.1093/nar/15.7.2971. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Simon J. A., Sutton C. A., Lobell R. B., Glaser R. L., Lis J. T. Determinants of heat shock-induced chromosome puffing. Cell. 1985 Apr;40(4):805–817. doi: 10.1016/0092-8674(85)90340-x. [DOI] [PubMed] [Google Scholar]
- Steller H., Pirrotta V. Expression of the Drosophila white gene under the control of the hsp70 heat shock promoter. EMBO J. 1985 Dec 30;4(13B):3765–3772. doi: 10.1002/j.1460-2075.1985.tb04146.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Struhl G. Near-reciprocal phenotypes caused by inactivation or indiscriminate expression of the Drosophila segmentation gene ftz. Nature. 1985 Dec 19;318(6047):677–680. doi: 10.1038/318677a0. [DOI] [PubMed] [Google Scholar]
- Tabor S., Richardson C. C. DNA sequence analysis with a modified bacteriophage T7 DNA polymerase. Proc Natl Acad Sci U S A. 1987 Jul;84(14):4767–4771. doi: 10.1073/pnas.84.14.4767. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Tamura T., Kunert C., Postlethwait J. Sex- and cell-specific regulation of yolk polypeptide genes introduced into Drosophila by P-element-mediated gene transfer. Proc Natl Acad Sci U S A. 1985 Oct;82(20):7000–7004. doi: 10.1073/pnas.82.20.7000. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Topol J., Ruden D. M., Parker C. S. Sequences required for in vitro transcriptional activation of a Drosophila hsp 70 gene. Cell. 1985 Sep;42(2):527–537. doi: 10.1016/0092-8674(85)90110-2. [DOI] [PubMed] [Google Scholar]
- Velazquez J. M., Sonoda S., Bugaisky G., Lindquist S. Is the major Drosophila heat shock protein present in cells that have not been heat shocked? J Cell Biol. 1983 Jan;96(1):286–290. doi: 10.1083/jcb.96.1.286. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Walder J. Antisense DNA and RNA: progress and prospects. Genes Dev. 1988 May;2(5):502–504. doi: 10.1101/gad.2.5.502. [DOI] [PubMed] [Google Scholar]
- Warren T. G., Brennan M. D., Mahowald A. P. Two processing steps in maturation of vitellogenin polypeptides in Drosophila melanogaster. Proc Natl Acad Sci U S A. 1979 Jun;76(6):2848–2852. doi: 10.1073/pnas.76.6.2848. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Williams J. L., Bownes M. Reduced stability of RNA coding for yolk polypeptide 3 in Drosophila melanogaster ovary. Eur J Biochem. 1986 Nov 17;161(1):95–101. doi: 10.1111/j.1432-1033.1986.tb10128.x. [DOI] [PubMed] [Google Scholar]
- Xiao H., Lis J. T. A consensus sequence polymer inhibits in vivo expression of heat shock genes. Mol Cell Biol. 1986 Sep;6(9):3200–3206. doi: 10.1128/mcb.6.9.3200. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Xiao H., Lis J. T. Germline transformation used to define key features of heat-shock response elements. Science. 1988 Mar 4;239(4844):1139–1142. doi: 10.1126/science.3125608. [DOI] [PubMed] [Google Scholar]
- Yost H. J., Lindquist S. RNA splicing is interrupted by heat shock and is rescued by heat shock protein synthesis. Cell. 1986 Apr 25;45(2):185–193. doi: 10.1016/0092-8674(86)90382-x. [DOI] [PubMed] [Google Scholar]