The Drosophila PAR Domain Protein 1 (Pdp1) Gene Encodes Multiple Differentially Expressed mRNAs and Proteins through the Use of Multiple Enhancers and Promoters

Abstract

Transcription factors are often expressed at several times and in multiple tissues during development and regulate diverse sets of downstream target genes by varying their combinatorial interactions with other transcription factors. The Drosophila Tropomyosin I (TmI) gene is regulated by a complex of proteins within the enhancer that synergistically interacts with MEF2 to activate TmI transcription as muscle cells fuse and differentiate. One of the components of this complex is PDP1 (PAR domain protein 1), a basic leucine zipper transcription factor that is highly homologous to three vertebrate genes that are members of the PAR domain subfamily. We have isolated and describe here the structure of the Pdp1 gene. The Pdp1 gene is complex, containing at least four transcriptional start sites and producing at least six different mRNAs and PDP1 isoforms. Five of the PDP1 isoforms differ by the substitution or insertion of amino acids at or near the N-terminal of the protein. At least three of these alternately spliced transcripts are differentially expressed in different tissues of the developing embryo in which PDP1 expression is correlated with the differentiation of different cell types. A sixth isoform is produced by splicing out part of the PAR and basic DNA binding domains, and DNA binding and transient transfection experiments suggest that it functions as a dominant negative inhibitor of transcription. Furthermore, two enhancers have been identified within the gene that express in the somatic mesodermal precursors to body wall muscles and fat body and together direct expression in other tissues that closely mimics that of the endogenous gene. These results show that Pdp1 is widely expressed, including in muscle, fat, and gut precursors, and is likely involved in the transcriptional control of different developmental pathways through the use of differentially expressed PDP1 isoforms. Furthermore, the similarities between Pdp1 and the other PAR domain genes suggest that Pdp1 is the homologue of the vertebrate genes.