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. 1988 Sep;120(1):173–180. doi: 10.1093/genetics/120.1.173

Analysis of the Promoter of the Rh2 Opsin Gene in Drosophila Melanogaster

D Mismer 1, W M Michael 1, T R Laverty 1, G M Rubin 1
PMCID: PMC1203488  PMID: 2975615

Abstract

We have analyzed the cis-acting regulatory sequences of the Drosophila melanogaster Rh2 gene that encodes the protein component of a rhodopsin which is expressed in ocellar photoreceptor cells. DNA fragments containing the start point of transcription of the Rh2 gene were fused to either the Escherichia coli chloramphenicol acetyltransferase (CAT) or lacZ (β-galactosidase) genes and introduced into the Drosophila germline by P-element-mediated transformation. Expression of the E. coli genes was then used to assay the ability of various sequences from the Rh2 gene to confer upon the indicator genes the Rh2 pattern of expression. Fragments containing between 4.3 kb and 183 bp upstream of the start of transcription plus the first 32 bp of the 5'-untranslated leader were found to result in nearly identical levels of head-specific CAT expression. Deletion of Rh2 sequences distal to position -112 bp resulted in loss of detectable CAT expression from these Rh2/CAT fusion constructs. We have, therefore, defined a region essential for head-specific expression of the Rh2 gene to a region extending from -183 to -112. We have determined the DNA sequence of the Rh2 promoter from -448 to +32 and have found an 11-bp sequence which is also present in the upstream flanking sequences of two other photoreceptor-specific genes (ninaE and ninaC). By histochemical staining of β-galactosidase expressed under the control of the Rh2 promoter and by analyzing the effect of the ocelliless mutation on the expression of an Rh2/CAT fusion gene, we have been able to demonstrate that this promoter is active in ocelli.

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Selected References

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  1. Bourouis M., Richards G. Remote regulatory sequences of the Drosophila glue gene sgs3 as revealed by P-element transformation. Cell. 1985 Feb;40(2):349–357. doi: 10.1016/0092-8674(85)90149-7. [DOI] [PubMed] [Google Scholar]
  2. Cohen R. S., Meselson M. Separate regulatory elements for the heat-inducible and ovarian expression of the Drosophila hsp26 gene. Cell. 1985 Dec;43(3 Pt 2):737–746. doi: 10.1016/0092-8674(85)90247-8. [DOI] [PubMed] [Google Scholar]
  3. Cowman A. F., Zuker C. S., Rubin G. M. An opsin gene expressed in only one photoreceptor cell type of the Drosophila eye. Cell. 1986 Mar 14;44(5):705–710. doi: 10.1016/0092-8674(86)90836-6. [DOI] [PubMed] [Google Scholar]
  4. Delaney S. J., Sunkel C. E., Genova-Seminova G. K., Davies J. E., Glover D. M. Cis-acting sequences sufficient for correct tissue and temporal specificity of larval serum protein 1 genes of Drosophila. EMBO J. 1987 Dec 1;6(12):3849–3854. doi: 10.1002/j.1460-2075.1987.tb02722.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  5. 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]
  6. Feiler R., Harris W. A., Kirschfeld K., Wehrhahn C., Zuker C. S. Targeted misexpression of a Drosophila opsin gene leads to altered visual function. Nature. 1988 Jun 23;333(6175):737–741. doi: 10.1038/333737a0. [DOI] [PubMed] [Google Scholar]
  7. Fryxell K. J., Meyerowitz E. M. An opsin gene that is expressed only in the R7 photoreceptor cell of Drosophila. EMBO J. 1987 Feb;6(2):443–451. doi: 10.1002/j.1460-2075.1987.tb04774.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  8. 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]
  9. 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]
  10. Gorman C. M., Moffat L. F., Howard B. H. Recombinant genomes which express chloramphenicol acetyltransferase in mammalian cells. Mol Cell Biol. 1982 Sep;2(9):1044–1051. doi: 10.1128/mcb.2.9.1044. [DOI] [PMC free article] [PubMed] [Google Scholar]
  11. Hazelrigg T., Levis R., Rubin G. M. Transformation of white locus DNA in drosophila: dosage compensation, zeste interaction, and position effects. Cell. 1984 Feb;36(2):469–481. doi: 10.1016/0092-8674(84)90240-x. [DOI] [PubMed] [Google Scholar]
  12. Hiromi Y., Kuroiwa A., Gehring W. J. Control elements of the Drosophila segmentation gene fushi tarazu. Cell. 1985 Dec;43(3 Pt 2):603–613. doi: 10.1016/0092-8674(85)90232-6. [DOI] [PubMed] [Google Scholar]
  13. Jowett T. The regulatory domain of a larval serum protein gene in Drosophila melanogaster. EMBO J. 1985 Dec 30;4(13B):3789–3795. doi: 10.1002/j.1460-2075.1985.tb04149.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  14. Kalfayan L., Wakimoto B., Spradling A. Analysis of transcriptional regulation of the s38 chorion gene of Drosophila by P element-mediated transformation. J Embryol Exp Morphol. 1984 Nov;83 (Suppl):137–146. [PubMed] [Google Scholar]
  15. Krumm A., Roth G. E., Korge G. Transformation of salivary gland secretion protein gene Sgs-4 in Drosophila: stage- and tissue-specific regulation, dosage compensation, and position effect. Proc Natl Acad Sci U S A. 1985 Aug;82(15):5055–5059. doi: 10.1073/pnas.82.15.5055. [DOI] [PMC free article] [PubMed] [Google Scholar]
  16. Langer-Safer P. R., Levine M., Ward D. C. Immunological method for mapping genes on Drosophila polytene chromosomes. Proc Natl Acad Sci U S A. 1982 Jul;79(14):4381–4385. doi: 10.1073/pnas.79.14.4381. [DOI] [PMC free article] [PubMed] [Google Scholar]
  17. Levine J., Spradling A. DNA sequence of a 3.8 kilobase pair region controlling Drosophila chorion gene amplification. Chromosoma. 1985;92(2):136–142. doi: 10.1007/BF00328465. [DOI] [PubMed] [Google Scholar]
  18. Levis R., Hazelrigg T., Rubin G. M. Separable cis-acting control elements for expression of the white gene of Drosophila. EMBO J. 1985 Dec 16;4(13A):3489–3499. doi: 10.1002/j.1460-2075.1985.tb04108.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  19. Marsh J. L., Gibbs P. D., Timmons P. M. Developmental control of transduced dopa decarboxylase genes in D. melanogaster. Mol Gen Genet. 1985;198(3):393–403. doi: 10.1007/BF00332929. [DOI] [PubMed] [Google Scholar]
  20. Mestril R., Schiller P., Amin J., Klapper H., Ananthan J., Voellmy R. Heat shock and ecdysterone activation of the Drosophila melanogaster hsp23 gene; a sequence element implied in developmental regulation. EMBO J. 1986 Jul;5(7):1667–1673. doi: 10.1002/j.1460-2075.1986.tb04410.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  21. Mismer D., Rubin G. M. Analysis of the promoter of the ninaE opsin gene in Drosophila melanogaster. Genetics. 1987 Aug;116(4):565–578. doi: 10.1093/genetics/116.4.565. [DOI] [PMC free article] [PubMed] [Google Scholar]
  22. Montell C., Rubin G. M. The Drosophila ninaC locus encodes two photoreceptor cell specific proteins with domains homologous to protein kinases and the myosin heavy chain head. Cell. 1988 Mar 11;52(5):757–772. doi: 10.1016/0092-8674(88)90413-8. [DOI] [PubMed] [Google Scholar]
  23. Nathans J., Hogness D. S. Isolation and nucleotide sequence of the gene encoding human rhodopsin. Proc Natl Acad Sci U S A. 1984 Aug;81(15):4851–4855. doi: 10.1073/pnas.81.15.4851. [DOI] [PMC free article] [PubMed] [Google Scholar]
  24. 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]
  25. Nathans J., Thomas D., Hogness D. S. Molecular genetics of human color vision: the genes encoding blue, green, and red pigments. Science. 1986 Apr 11;232(4747):193–202. doi: 10.1126/science.2937147. [DOI] [PubMed] [Google Scholar]
  26. Ovchinnikov YuA Rhodopsin and bacteriorhodopsin: structure-function relationships. FEBS Lett. 1982 Nov 8;148(2):179–191. doi: 10.1016/0014-5793(82)80805-3. [DOI] [PubMed] [Google Scholar]
  27. 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]
  28. 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]
  29. Pirrotta V., Steller H., Bozzetti M. P. Multiple upstream regulatory elements control the expression of the Drosophila white gene. EMBO J. 1985 Dec 16;4(13A):3501–3508. doi: 10.1002/j.1460-2075.1985.tb04109.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  30. Pollock J. A., Benzer S. Transcript localization of four opsin genes in the three visual organs of Drosophila; RH2 is ocellus specific. Nature. 1988 Jun 23;333(6175):779–782. doi: 10.1038/333779a0. [DOI] [PubMed] [Google Scholar]
  31. Riddihough G., Pelham H. R. Activation of the Drosophila hsp27 promoter by heat shock and by ecdysone involves independent and remote regulatory sequences. EMBO J. 1986 Jul;5(7):1653–1658. doi: 10.1002/j.1460-2075.1986.tb04408.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  32. 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]
  33. Scholnick S. B., Bray S. J., Morgan B. A., McCormick C. A., Hirsh J. CNS and hypoderm regulatory elements of the Drosophila melanogaster dopa decarboxylase gene. Science. 1986 Nov 21;234(4779):998–1002. doi: 10.1126/science.3095924. [DOI] [PubMed] [Google Scholar]
  34. Scholnick S. B., Morgan B. A., Hirsh J. The cloned dopa decarboxylase gene is developmentally regulated when reintegrated into the Drosophila genome. Cell. 1983 Aug;34(1):37–45. doi: 10.1016/0092-8674(83)90134-4. [DOI] [PubMed] [Google Scholar]
  35. 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]
  36. Spradling A. C., Rubin G. M. The effect of chromosomal position on the expression of the Drosophila xanthine dehydrogenase gene. Cell. 1983 Aug;34(1):47–57. doi: 10.1016/0092-8674(83)90135-6. [DOI] [PubMed] [Google Scholar]
  37. Wu C. Activating protein factor binds in vitro to upstream control sequences in heat shock gene chromatin. Nature. 1984 Sep 6;311(5981):81–84. doi: 10.1038/311081a0. [DOI] [PubMed] [Google Scholar]
  38. Wu C. An exonuclease protection assay reveals heat-shock element and TATA box DNA-binding proteins in crude nuclear extracts. Nature. 1985 Sep 5;317(6032):84–87. doi: 10.1038/317084a0. [DOI] [PubMed] [Google Scholar]
  39. Zuker C. S., Mismer D., Hardy R., Rubin G. M. Ectopic expression of a minor Drosophila opsin in the major photoreceptor cell class: distinguishing the role of primary receptor and cellular context. Cell. 1988 May 6;53(3):475–482. doi: 10.1016/0092-8674(88)90167-5. [DOI] [PubMed] [Google Scholar]
  40. Zuker C. S., Montell C., Jones K., Laverty T., Rubin G. M. A rhodopsin gene expressed in photoreceptor cell R7 of the Drosophila eye: homologies with other signal-transducing molecules. J Neurosci. 1987 May;7(5):1550–1557. doi: 10.1523/JNEUROSCI.07-05-01550.1987. [DOI] [PMC free article] [PubMed] [Google Scholar]

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