Abstract
The pivotal role of twist in mesoderm determination in the Drosophila embryo depends upon two processes--the transcriptional activation of twist in the ventrally located mesodermal anlage and the regulation of downstream gene expression by the twist transcription factor. To elucidate the molecular mechanisms involved in these processes, we have compared both the coding and regulatory regions of the twist genes from Drosophila melanogaster and Drosophila virilis. Within the coding region, the basic-helix-loop-helix DNA binding and dimerization motif is highly conserved, consistent with the functional importance of this domain. A comparison of the transcriptional regulatory regions reveals a high degree of conservation in the more distal of the two ventral activator regions that have been mapped in the twist 5' flanking region. On the other hand, the more proximal ventral activator region is absent at the corresponding position in the D. virilis twist gene. Instead, there is a region in the second intron of the D. virilis gene that resembles the proximal element of the D. melanogaster gene, in that it consists of little more than a series of whole and half binding sites for the dorsal morphogen. In transformation experiments, the intronic D. virilis element directs an expression pattern that is indistinguishable from that directed by the D. melanogaster proximal VAR. Thus, the twi genes from these two species appear to have evolved enhancer elements with very similar structural and functional properties. These findings suggest that apparently redundant spatially regulated enhancer elements may each play essential roles in fine tuning the level and/or pattern of gene expression.
