PROMOTER IS AN IMPORTANT DETERMINANT OF DEVELOPMENTALLY REGULATED PUFFING AT THE SGS-4 LOCUS OF DROSOPHILA-MELANOGASTER

Sgs-4 is one of the eight known genes coding for larval secretion proteins in Drosophila melanogaster. High-level transcription of the endogenous Sgs genes in salivary glands is accompanied by chromosome puffing at the Sgs gene loci. Naturally occurring mutations of the Sgs-4 promoter region diminish both the level of Sgs-4 expression and the puff size; in null-producers no puff is formed. P element-mediated transformation experiments were performed to clarify this apparent causal relation between transcription and puffing. Sgs-4 upstream sequences, unchanged or recombined with sequences from differently expressed alleles, were fused with Sgs-4 coding and downstream sequences or with the coding sequence of the viral oncogene v-mil. Analyses of the expression of these fragments at the RNA and protein levels and of their capacity for puff formation demonstrate uncoupling of transcription and puffing. That is, high-level transcription is independent of chromosome puffing and does not necessarily induce puffing, and developmentally regulated chromosome puffing is independent of significant transcriptional activity within the puff. Our results show that the strength of the Sgs-4 promoter located within the upstream region from -1 to -840 determines the formation of a puff. No specific effects could be detected on either transcription or puffing by decondensed versus compact chromatin adjoining the transposed DNA at the sites of insertion in transformants. A model in which trans-acting factors binding to the promoter region initiate puffing is proposed. © 1990.