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EMBO J. Feb 1993; 12(2): 527–535.
PMCID: PMC413235

The Drosophila doublesex proteins share a novel zinc finger related DNA binding domain.

Abstract

The doublesex gene of Drosophila melanogaster is the final member of a well characterized hierarchy of genes that controls somatic sex determination and differentiation. The male-specific and female-specific doublesex polypeptides occupy a terminal position in the hierarchy, and thus regulate those genes responsible for the development of sexually dimorphic characteristics of the fly. To investigate the molecular mechanism by which these two related proteins interact with specific target genes, we have identified and characterized their DNA binding domains. Using gel mobility shift experiments with sequentially deleted polypeptides, site-directed mutagenesis and spectrophotometric assays, we have shown that the two doublesex proteins share a common and novel zinc finger-related DNA binding domain distinct from any reported class of zinc binding proteins. We have further shown that of 10 null dsx alleles, six encode proteins deficient in DNA binding activity, and that three of these alleles are the result of mutations that alter cysteine and histidine residues in the metal binding domain. Our results provide evidence that both the male-specific and female-specific doublesex proteins share and depend upon the same DNA binding domain for function in vivo, suggesting that both proteins bind to, but differentially regulate, a common set of genes in both sexes.

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Selected References

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  • Ota T, Fukunaga A, Kawabe M, Oishi K. Interactions between sex-transformation mutants of Drosophila melanogaster. I. Hemolymph vitellogenins and gonad morphology. Genetics. 1981 Nov-Dec;99(3-4):429–441. [PMC free article] [PubMed]
  • Postlethwait JH, Bownes M, Jowett T. Sexual phenotype and vitellogenin synthesis in Drosophila melanogaster. Dev Biol. 1980 Oct;79(2):379–387. [PubMed]
  • Schena M, Freedman LP, Yamamoto KR. Mutations in the glucocorticoid receptor zinc finger region that distinguish interdigitated DNA binding and transcriptional enhancement activities. Genes Dev. 1989 Oct;3(10):1590–1601. [PubMed]
  • Severne Y, Wieland S, Schaffner W, Rusconi S. Metal binding 'finger' structures in the glucocorticoid receptor defined by site-directed mutagenesis. EMBO J. 1988 Aug;7(8):2503–2508. [PMC free article] [PubMed]
  • Slee R, Bownes M. Sex determination in Drosophila melanogaster. Q Rev Biol. 1990 Jun;65(2):175–204. [PubMed]
  • Smith DL, Johnson AD. A molecular mechanism for combinatorial control in yeast: MCM1 protein sets the spacing and orientation of the homeodomains of an alpha 2 dimer. Cell. 1992 Jan 10;68(1):133–142. [PubMed]
  • Steinmann-Zwicky M, Amrein H, Nöthiger R. Genetic control of sex determination in Drosophila. Adv Genet. 1990;27:189–237. [PubMed]
  • Tabor S, Richardson CC. A bacteriophage T7 RNA polymerase/promoter system for controlled exclusive expression of specific genes. Proc Natl Acad Sci U S A. 1985 Feb;82(4):1074–1078. [PMC free article] [PubMed]
  • Taylor BJ, Truman JW. Commitment of abdominal neuroblasts in Drosophila to a male or female fate is dependent on genes of the sex-determining hierarchy. Development. 1992 Mar;114(3):625–642. [PubMed]
  • Vallee BL, Coleman JE, Auld DS. Zinc fingers, zinc clusters, and zinc twists in DNA-binding protein domains. Proc Natl Acad Sci U S A. 1991 Feb 1;88(3):999–1003. [PMC free article] [PubMed]
  • Webster LC, Zhang K, Chance B, Ayene I, Culp JS, Huang WJ, Wu FY, Ricciardi RP. Conversion of the E1A Cys4 zinc finger to a nonfunctional His2,Cys2 zinc finger by a single point mutation. Proc Natl Acad Sci U S A. 1991 Nov 15;88(22):9989–9993. [PMC free article] [PubMed]
  • Wolfner MF. Sex-specific gene expression in somatic tissues of Drosophila melanogaster. Trends Genet. 1988 Dec;4(12):333–337. [PubMed]
  • Baker BS. Sex in flies: the splice of life. Nature. 1989 Aug 17;340(6234):521–524. [PubMed]
  • Baker BS, Ridge KA. Sex and the single cell. I. On the action of major loci affecting sex determination in Drosophila melanogaster. Genetics. 1980 Feb;94(2):383–423. [PMC free article] [PubMed]
  • Baker BS, Hoff G, Kaufman TC, Wolfner MF, Hazelrigg T. The doublesex locus of Drosophila melanogaster and its flanking regions: a cytogenetic analysis. Genetics. 1991 Jan;127(1):125–138. [PMC free article] [PubMed]
  • Belote JM, Handler AM, Wolfner MF, Livak KJ, Baker BS. Sex-specific regulation of yolk protein gene expression in Drosophila. Cell. 1985 Feb;40(2):339–348. [PubMed]
  • Berg JM. Potential metal-binding domains in nucleic acid binding proteins. Science. 1986 Apr 25;232(4749):485–487. [PubMed]
  • Blumberg H, Eisen A, Sledziewski A, Bader D, Young ET. Two zinc fingers of a yeast regulatory protein shown by genetic evidence to be essential for its function. Nature. 328(6129):443–445. [PubMed]
  • Bownes M, Nöthiger R. Sex determining genes and vitellogenin synthesis in Drosophila melanogaster. Mol Gen Genet. 1981;182(2):222–228. [PubMed]
  • Burtis KC, Baker BS. Drosophila doublesex gene controls somatic sexual differentiation by producing alternatively spliced mRNAs encoding related sex-specific polypeptides. Cell. 1989 Mar 24;56(6):997–1010. [PubMed]
  • Burtis KC, Coschigano KT, Baker BS, Wensink PC. The doublesex proteins of Drosophila melanogaster bind directly to a sex-specific yolk protein gene enhancer. EMBO J. 1991 Sep;10(9):2577–2582. [PMC free article] [PubMed]
  • Chou PY, Fasman GD. Prediction of the secondary structure of proteins from their amino acid sequence. Adv Enzymol Relat Areas Mol Biol. 1978;47:45–148. [PubMed]
  • Clark JM. Novel non-templated nucleotide addition reactions catalyzed by procaryotic and eucaryotic DNA polymerases. Nucleic Acids Res. 1988 Oct 25;16(20):9677–9686. [PMC free article] [PubMed]
  • Cline TW. The affairs of daughterless and the promiscuity of developmental regulators. Cell. 1989 Oct 20;59(2):231–234. [PubMed]
  • Devereux J, Haeberli P, Smithies O. A comprehensive set of sequence analysis programs for the VAX. Nucleic Acids Res. 1984 Jan 11;12(1 Pt 1):387–395. [PMC free article] [PubMed]
  • Frankel AD, Kim PS. Modular structure of transcription factors: implications for gene regulation. Cell. 1991 May 31;65(5):717–719. [PubMed]
  • Guan KL, Dixon JE. Eukaryotic proteins expressed in Escherichia coli: an improved thrombin cleavage and purification procedure of fusion proteins with glutathione S-transferase. Anal Biochem. 1991 Feb 1;192(2):262–267. [PubMed]
  • Harrison SC. A structural taxonomy of DNA-binding domains. Nature. 1991 Oct 24;353(6346):715–719. [PubMed]
  • HILDRETH PE. DOUBLESEX, RECESSIVE GENE THAT TRANSFORMS BOTH MALES AND FEMALES OF DROSOPHILA INTO INTERSEXES. Genetics. 1965 Apr;51:659–678. [PMC free article] [PubMed]
  • Hunt JB, Neece SH, Ginsburg A. The use of 4-(2-pyridylazo)resorcinol in studies of zinc release from Escherichia coli aspartate transcarbamoylase. Anal Biochem. 1985 Apr;146(1):150–157. [PubMed]
  • Jones N. Transcriptional regulation by dimerization: two sides to an incestuous relationship. Cell. 1990 Apr 6;61(1):9–11. [PubMed]
  • Luisi B. DNA transcription. Zinc standard for economy. Nature. 1992 Apr 2;356(6368):379–380. [PubMed]
  • Nagoshi RN, Baker BS. Regulation of sex-specific RNA splicing at the Drosophila doublesex gene: cis-acting mutations in exon sequences alter sex-specific RNA splicing patterns. Genes Dev. 1990 Jan;4(1):89–97. [PubMed]
  • Nagoshi RN, McKeown M, Burtis KC, Belote JM, Baker BS. The control of alternative splicing at genes regulating sexual differentiation in D. melanogaster. Cell. 1988 Apr 22;53(2):229–236. [PubMed]

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