Contrasting activities of the aggregative and late PDSA promoters in Dictyostelium development.
Expression of the Dictyostelium PdsA gene from the aggregative (PdA) and late (PdL) promoter is essential for aggregation and slug morphogenesis, respectively. We studied the regulation of the PdA and PdL promoters in slugs using labile beta-galactosidase (gal) reporter enzymes. PdL was active in prestalk cells as was also found with stable gal. PdA activity decreased strongly in slugs from all cells, except those at the rear. This is almost opposite to PdA activity traced with stable gal, where slugs showed sustained activity with highest levels at the front. PdA was down-regulated after aggregation irrespective of stimulation with any of the factors known to control gene expression. PdL activity was induced in cell suspension by cAMP and DIF acting in synergy. However, a DIF-less mutant showed normal PdL activity during development, suggesting that DIF does not control PdL in vivo. Dissection of the PdL promoter showed that all sequences essential for correct spatiotemporal control of promoter activity are downstream of the transcription start site in a region between -383 and -19 nucleotides relative to the start codon. Removal of nucleotides to position -364 eliminated responsiveness to DIF and cAMP, but normal PdL activity in prestalk cells in slugs was retained. Further 5' deletions abolished all promoter activity. This result also indicates that the induction by DIF and cAMP as seen in cell suspensions is not essential for PdL activity in normal development.
Weening
Karin E
KE
School of Life Sciences, University of Dundee, Dundee, DD1 5EH, UK.
Wijk
Irene Verkerke-Van
IV
Thompson
Christopher R
CR
Kessin
Richard H
RH
Podgorski
Gregory J
GJ
Schaap
Pauline
P
D015105:3',5'-Cyclic-AMP Phosphodiesterases / Q000235:genetics; D000818:Animals; D001483:Base Sequence; D000242:Cyclic AMP / Q000494:pharmacology; D016054:DNA, Protozoan / Q000235:genetics; D004023:Dictyostelium / Q000201:enzymology / Q000235:genetics* / Q000254:growth & development*; D015536:Down-Regulation / Q000187:drug effects; D018507:Gene Expression Regulation, Developmental / Q000187:drug effects; D017125:Genes, Protozoan; D017930:Genes, Reporter; D006588:Hexanones / Q000494:pharmacology; D008969:Molecular Sequence Data; D011401:Promoter Regions, Genetic; D017384:Sequence Deletion; D015398:Signal Transduction; D015854:Up-Regulation / Q000187:drug effects
D016054:DNA, Protozoan; D006588:Hexanones; C053433:1-((3,5-dichloro)-2,6-dihydroxy-4-methoxyphenyl)-1-hexanone; D000242:Cyclic AMP; D015105:3',5'-Cyclic-AMP Phosphodiesterases