GATA-type zinc finger motif-containing sequences and chorion gene transcription factors of the silkworm Bombyx mori.

Abstract

To characterize a DNA-binding protein, BCFI, which regulates the expression of silkmoth chorion genes through binding to gene promoter elements identical to those recognized by the GATA family of transcription factors, we have carried out polymerase chain reaction amplifications of Bombyx mori genomic DNA using degenerate primers derived from the conserved DNA binding domain of mammalian GATA factors. Two single copy genes, BmGATA alpha and BmGATA beta, were identified, which encode sequences containing GATA-type zinc finger motifs. The BmGATA beta gene is expressed in follicular and Bm5 tissue culture cells, the two cell types that contain BCFI. No BmGATA alpha gene transcripts were detectable in the tissues that were tested. Upon overexpression in Escherichia coli, a peptide encompassing the BmGATA beta zinc finger motif was able to bind specifically to the BCFI recognition motif of the chorion gene promoters. A polyclonal antibody directed against the zinc finger domain of BmGATA beta was also used in gel retardation assays to confirm that factor BCFI is indeed encoded by the BmGATA beta gene. Conceptual translation of a complete cDNA clone encoding the BmGATA beta protein revealed that this protein has a size similar to that of an immunoreactive protein, presumably BCFI, which is present in follicular cell extracts.