Regulation of sexual dimorphism

Mutational and chemogenetic analysis of the doublesex DM domain

Wei Zhang, Biaoru Li, Rupinder Singh, Uma Narendra, Lingyang Zhu, Michael A. Weiss

Research output: Contribution to journalArticle

17 Citations (Scopus)

Abstract

Doublesex (dsx) is a transcription factor in Drosophila that regulates somatic sexual differentiation. Male- and female-speciftc splicing isoforms of DSX share a novel DNA-binding domain, designated the DM motif. Broadly conserved among metazoan sex-determining factors, the DM domain contains a nonclassical zinc module and binds in the DNA minor groove. Here, we characterize the DM motif by site-directed and random mutagenesis using a yeast one-hybrid (Y1H) system and extend this analysis by chemogenetic complementation in vitro. The Y1H system is based on a sex-speciftc Drosophila enhancer element and validated through studies of intersexual dsx mutations. We demonstrate that the eight motif-specific histidines and cysteines engaged in zinc coordination are each critical and cannot be interchanged; folding also requires conserved aliphatic side chains in the hydrophobic core. Mutations that impair DNA binding tend to occur at conserved positions, whereas neutral substitutions occur at nonconserved sites. Evidence for a specific salt bridge between a conserved lysine and the DNA backbone is obtained through the synthesis of nonstandard protein and DNA analogs. Together, these results provide molecular links between the structure of the DM domain and its function in the regulation of sexual dimorphism.

Original languageEnglish (US)
Pages (from-to)535-547
Number of pages13
JournalMolecular and Cellular Biology
Volume26
Issue number2
DOIs
StatePublished - Jan 1 2006

Fingerprint

Sex Characteristics
DNA
Drosophila
Zinc
Sex Factors
Sex Differentiation
Mutation
Site-Directed Mutagenesis
Histidine
Cysteine
Protein Isoforms
Transcription Factors
Salts
Yeasts
Proteins

ASJC Scopus subject areas

  • Molecular Biology
  • Cell Biology

Cite this

Regulation of sexual dimorphism : Mutational and chemogenetic analysis of the doublesex DM domain. / Zhang, Wei; Li, Biaoru; Singh, Rupinder; Narendra, Uma; Zhu, Lingyang; Weiss, Michael A.

In: Molecular and Cellular Biology, Vol. 26, No. 2, 01.01.2006, p. 535-547.

Research output: Contribution to journalArticle

Zhang, Wei ; Li, Biaoru ; Singh, Rupinder ; Narendra, Uma ; Zhu, Lingyang ; Weiss, Michael A. / Regulation of sexual dimorphism : Mutational and chemogenetic analysis of the doublesex DM domain. In: Molecular and Cellular Biology. 2006 ; Vol. 26, No. 2. pp. 535-547.
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