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The Dd ras gene produces three transcripts during Dictyostelium development. The largest transcript (L-) can be induced by external addition of cAMP even in cells prevented from aggregating, whereas shorter transcripts (S1- and S2-) expression requires cell aggregate formation. We show the presence of two independent promoters for L- and S-transcripts by deletion analysis of Dd ras fragments fused to CAT reporter genes reintroduced in Dictyostelium. A direct repeat upstream of S-transcript start sites which seems involved in S-promoter function, modulates also L-RNA accumulation. Furthermore removal of sequences between this repeat and the AUG protein start codon reduces the level of L-transcripts in aggregates. This study allowed to uncover the intricate pattern of sequences participating in the regulation of Dd ras expression.

Two Dictyostelium discoideum protein kinase(PK)-encoding cDNAs (Dd PK1 and Dd PK2) have been isolated by hybridization with an oligodeoxyribonucleotide derived from a highly conserved region of eukaryotic PKs. The two nucleotide (nt) sequences encode new putative serine/threonine-specific PKs. Dd PK1 is a partial cDNA covering the entire catalytic domain. The derived amino acid (aa) sequence is about 30% identical to both cAMP-dependent protein kinase (cAPK) and protein kinase C. The Dd PK2 sequence was extended through the isolation of a genomic fragment encoding a complete putative protein. A single intron is present, as deduced from sequence comparison with the cDNA. The catalytic domain appears more closely related to the catalytic subunit of cAPK (54% sequence identity). However, our nt sequence potentially codes for a much larger protein (648 vs. about 350 aa for most cAPKs) with a N-terminal half containing long homopolymers of threonines, glutamines and asparagines. Similar repeats occur at the C terminus of Dd PK1, Dd PK1 is expressed in vegetatively growing cells and during development. Dd PK1 RNA decreases after 6 h of starvation to re-accumulate once the cells have aggregated. Dd PK2 transcripts, present at a low amount in growing cells, rise upon starvation. A switch to a shorter form of transcripts occurs between 3 and 6 h into development.