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PMID:10717484
Citation |
Zhu, W and Hanes, SD (2000) Identification of drosophila bicoid-interacting proteins using a custom two-hybrid selection. Gene 245:329-39 |
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Abstract |
Bicoid directs pattern formation in the developing Drosophila embryo, and does so by performing two seemingly unrelated tasks; it activates transcription and represses translation. To understand how Bicoid carries out this dual role, we sought to identify Bicoid-ancillary proteins that might mediate Bicoid's function in transcription or translation. We used a customized version of the two-hybrid method and found two Bicoid-interacting proteins, Bin1 and Bin3, both of which interact with Bicoid in vitro. Bin1 is similar to a human protein (SAP18) involved in transcription regulation, and Bin3, described in this paper, is similar to a family of protein methyltransferases that modify RNA-binding proteins. Given that Bicoid's role as a translation regulator requires RNA binding, we suggest that the Bicoid-interacting methyltransferase might be important for that role. The custom two-hybrid method we used, in which Bicoid is bound to DNA via its own DNA binding domain, rather than via a fusion-protein tether, should be generally applicable to other DNA binding proteins. |
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Keywords |
Amino Acid Sequence; Animals; Carrier Proteins/genetics; Carrier Proteins/metabolism; Chromosome Mapping; DNA, Complementary/chemistry; DNA, Complementary/genetics; DNA, Complementary/isolation & purification; Drosophila/genetics; Drosophila/metabolism; Drosophila Proteins; Gene Expression Regulation, Developmental; Genetic Engineering; Homeodomain Proteins/genetics; Homeodomain Proteins/metabolism; Insect Proteins/genetics; Insect Proteins/metabolism; Methyltransferases/genetics; Methyltransferases/metabolism; Molecular Sequence Data; Protein Binding; Protein-Serine-Threonine Kinases; Recombinant Fusion Proteins/genetics; Recombinant Fusion Proteins/metabolism; Saccharomyces cerevisiae/genetics; Saccharomyces cerevisiae Proteins; Sequence Analysis, DNA; Sequence Homology, Amino Acid; Trans-Activators/genetics; Trans-Activators/metabolism; Transcription Factors; Two-Hybrid System Techniques |
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