Tumor-promoting phorbol esters (TPA and PDD) cause severe disorders in the growth and behavior of C. elegans. The
tpa-1 gene, which mediates the action of these agents, encodes a protein kinase C (PKC) homolog. The
tpa-1 gene consists of eleven exons and ten introns, and two mRNA species, 2.8 kb and 2.4 kb, are transcribed from the
tpa-1 locus. The former contains all the eleven exons and codes for TPA-1A, a protein of 80 kDa; the latter lacks the first four exons and encodes TPA-1B, a 65-kDa protein, without some 140 amino acids at the amino- terminus of TPA-1A. Microinjection rescue experiment, in which
tpa-1 genomic fragments variously lacking its 5' region were introduced into a TPA-resistant mutant strain, MJ563, showed that 1) TPA-1B was sufficient to confer phorbol ester- sensitivity on C. elegans; and 2) transcription for the 2.4- kb mRNA started at a region of the 4th intron located within 1 kb upstream from the 5th exon. We confirmed the promoter activity in this region by expressing lacZ fused to a genomic fragment covering only 1 kb upstream of the 5th exon. We also found with various
tpa-1-lacZ fusion genes that the tissue expression pattens of the two mRNAs were very different. These results suggest that the two PKC isoforms from
tpa-1 execute separate physiological functions in C. elegans. PKC is reproted to form a family of various isoforms in many animals. Recently, Y. Kohara et al. identified five cDNA clones encoding PKC homologs in their C. elegans cDNA sequencing project and kindly provided us with these clones. To understand the cellular signalling pathways that involve PKCs other than
tpa-1, we began to analyze these cDNA clones. We will discuss the current status of their structural analysis.