Synergistic Interactions of Silkmoth Chorion Promoter-Binding Factors

Abstract

Two DNA-binding proteins, BCFI and BCFII, that interact with defined promoter sequences of silkmoth chorion genes of late developmental specificity appear in the nuclei of follicular cells at a time that coincides with the transcriptional activation of the corresponding genes. BCFI prebinding is shown to be indispensable for stable binding of BCFII to its cognate sequence. BCFI and BCFII synergism requires a relatively stringent stereospecific alignment and is a prerequisite for the assembly of higher-order protein-promoter DNA complexes containing additional factors, which are neither gene (stage) nor class (chorion) specific. Binding of BCFI to its site correlates with the induction of DNA structural perturbations that may facilitate assembly of additional factors on the promoter. The BCFI-binding domain contains a core hexanucleotide sequence, AGATAA, which represents the major binding determinant of the erythroid-specific transcription factor GATA-1 of higher vertebrates. This sequence is shown to be necessary and sufficient for binding of BCFI, as it is for a factor that is present in induced K562 human erythroleukemic cells, presumably GATA-1. Comparative analyses of mobility shift patterns obtained with partially proteolyzed preparations of these two unrelated factors were used to confirm that a BCFI-like chorion promoter-binding protein, which is present in the nuclei of an established silkmoth cell line derived from ovarian tissue, is in fact BCFI. The transcriptional repression of endogenous chorion genes in this cell line coupled with the documented absence of factor BCFII suggests that the synergistic interactions between these two factors constitute a minimum requirement for late chorion gene expression.