Abstract
Previous experiments have identified a region that is required for the expression of the Drosophila glue protein gene Sgs-4 and is located 300-500 base pairs upstream from the structural gene. The chromatin in this region changes conformation and becomes hypersensitive to DNase I digestion when the gene becomes active, a change that apparently induces additional conformational changes near the site of transcription initiation. To learn more about the DNA sequence requirements for the function of this region, we analyzed three naturally occurring Sgs-4 under-producers. In two of these strains, a single base pair change within the hypersensitive region is correlated with a 50% reduction in the amount of Sgs-4 RNA produced. Another strain, which has multiple 5' lesions, is severely reduced in Sgs-4 expression and in the DNase hypersensitivity of the upstream region. Several of the sequence changes in this extreme underproducer lie near hypersensitive sites, suggesting that they inhibit the appearance of the normal DNase hypersensitive conformation.
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- Beckendorf S. K., Kafatos F. C. Differentiation in the salivary glands of Drosophila melanogaster: characterization of the glue proteins and their developmental appearance. Cell. 1976 Nov;9(3):365–373. doi: 10.1016/0092-8674(76)90081-7. [DOI] [PubMed] [Google Scholar]
- Benton W. D., Davis R. W. Screening lambdagt recombinant clones by hybridization to single plaques in situ. Science. 1977 Apr 8;196(4286):180–182. doi: 10.1126/science.322279. [DOI] [PubMed] [Google Scholar]
- Breathnach R., Chambon P. Organization and expression of eucaryotic split genes coding for proteins. Annu Rev Biochem. 1981;50:349–383. doi: 10.1146/annurev.bi.50.070181.002025. [DOI] [PubMed] [Google Scholar]
- Keene M. A., Corces V., Lowenhaupt K., Elgin S. C. DNase I hypersensitive sites in Drosophila chromatin occur at the 5' ends of regions of transcription. Proc Natl Acad Sci U S A. 1981 Jan;78(1):143–146. doi: 10.1073/pnas.78.1.143. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Korge G. Chromosome puff activity and protein synthesis in larval salivary glands of Drosophila melanogaster. Proc Natl Acad Sci U S A. 1975 Nov;72(11):4550–4554. doi: 10.1073/pnas.72.11.4550. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Korge G. Genetic analysis of the larval secretion gene Sgs-4 and its regulatory chromosome sites in Drosophila melanogaster. Chromosoma. 1981;84(3):373–390. doi: 10.1007/BF00286027. [DOI] [PubMed] [Google Scholar]
- Maxam A. M., Gilbert W. A new method for sequencing DNA. Proc Natl Acad Sci U S A. 1977 Feb;74(2):560–564. doi: 10.1073/pnas.74.2.560. [DOI] [PMC free article] [PubMed] [Google Scholar]
- McGhee J. D., Wood W. I., Dolan M., Engel J. D., Felsenfeld G. A 200 base pair region at the 5' end of the chicken adult beta-globin gene is accessible to nuclease digestion. Cell. 1981 Nov;27(1 Pt 2):45–55. doi: 10.1016/0092-8674(81)90359-7. [DOI] [PubMed] [Google Scholar]
- McGinnis W., Farrell J., Jr, Beckendorf S. K. Molecular limits on the size of a genetic locus in Drosophila melanogaster. Proc Natl Acad Sci U S A. 1980 Dec;77(12):7367–7371. doi: 10.1073/pnas.77.12.7367. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Murray N. E., Brammar W. J., Murray K. Lambdoid phages that simplify the recovery of in vitro recombinants. Mol Gen Genet. 1977 Jan 7;150(1):53–61. doi: 10.1007/BF02425325. [DOI] [PubMed] [Google Scholar]
- Muskavitch M. A., Hogness D. S. An expandable gene that encodes a Drosophila glue protein is not expressed in variants lacking remote upstream sequences. Cell. 1982 Jul;29(3):1041–1051. doi: 10.1016/0092-8674(82)90467-6. [DOI] [PubMed] [Google Scholar]
- Muskavitch M. A., Hogness D. S. Molecular analysis of a gene in a developmentally regulated puff of Drosophila melanogaster. Proc Natl Acad Sci U S A. 1980 Dec;77(12):7362–7366. doi: 10.1073/pnas.77.12.7362. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Rubin G. M., Spradling A. C. Genetic transformation of Drosophila with transposable element vectors. Science. 1982 Oct 22;218(4570):348–353. doi: 10.1126/science.6289436. [DOI] [PubMed] [Google Scholar]
- Shermoen A. W., Beckendorf S. K. A complex of interacting DNAase I-hypersensitive sites near the Drosophila glue protein gene, Sgs4. Cell. 1982 Jun;29(2):601–607. doi: 10.1016/0092-8674(82)90176-3. [DOI] [PubMed] [Google Scholar]
- Stalder J., Larsen A., Engel J. D., Dolan M., Groudine M., Weintraub H. Tissue-specific DNA cleavages in the globin chromatin domain introduced by DNAase I. Cell. 1980 Jun;20(2):451–460. doi: 10.1016/0092-8674(80)90631-5. [DOI] [PubMed] [Google Scholar]
- Thomas P. S. Hybridization of denatured RNA and small DNA fragments transferred to nitrocellulose. Proc Natl Acad Sci U S A. 1980 Sep;77(9):5201–5205. doi: 10.1073/pnas.77.9.5201. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Weinstock R., Sweet R., Weiss M., Cedar H., Axel R. Intragenic DNA spacers interrupt the ovalbumin gene. Proc Natl Acad Sci U S A. 1978 Mar;75(3):1299–1303. doi: 10.1073/pnas.75.3.1299. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Weintraub H., Larsen A., Groudine M. Alpha-Globin-gene switching during the development of chicken embryos: expression and chromosome structure. Cell. 1981 May;24(2):333–344. doi: 10.1016/0092-8674(81)90323-8. [DOI] [PubMed] [Google Scholar]
- Wu C., Gilbert W. Tissue-specific exposure of chromatin structure at the 5' terminus of the rat preproinsulin II gene. Proc Natl Acad Sci U S A. 1981 Mar;78(3):1577–1580. doi: 10.1073/pnas.78.3.1577. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Wu C. The 5' ends of Drosophila heat shock genes in chromatin are hypersensitive to DNase I. Nature. 1980 Aug 28;286(5776):854–860. doi: 10.1038/286854a0. [DOI] [PubMed] [Google Scholar]