Molecular cloning of a POU domain-containing factor involved in the regulation of the Bombyx sericin-1 gene.

Abstract

The POU domain is a highly conserved region found in a number of transcription factors and products of developmental control genes. We report here the isolation and characterization of a POU domain-containing cDNA (POU-M1) from the middle silk gland of Bombyx mori. It encodes a protein with a POU domain identical to that of the Drosophila Cf1-a protein. By mobility shift and nuclease protection assays, the POU-M1 protein and the putative silk gland factor 3 (SGF-3) were found to interact in an indistinguishable manner with the SC region (positions -204 to -183) of the sericin-1 gene, a key cis-acting element involved in the regulation of the gene through the interaction with SGF-3. Antibodies raised against the synthetic oligopeptide corresponding to the COOH-terminal region of the putative POU-M1 sequence reacted specifically to both the POU-M1 protein and SGF-3. Northern blot hybridization and Western blotting revealed that POU-M1 expression is regulated both temporally and spatially during silk gland development. We conclude that the POU-M1 protein is SGF-3 and propose that the differential expression of the POU-M1 gene is probably involved in the transcriptional regulation of the silk protein genes.