Multiple copies of a G-rich element upstream of a cAMP-inducible Dictyostelium gene are necessary but not sufficient for efficient gene expression.
The cysteine proteinase 1 (CP1) and cysteine proteinase 2 (CP2) genes of Dictyostelium discoideum encode co-ordinately expressed mRNA sequences which are inducible by extracellular cAMP. There are short, G-rich sequence elements upstream of both genes and we have previously shown that deletion of these elements from the CP2 gene abolishes cAMP-inducibility. We show here that the G-rich element from the CP1 gene is functionally homologous to that in the CP2 gene by reconstituting cAMP-inducibility in a deletion mutant of the CP2 gene using CP1-derived sequences. Both the CP1 and CP2 genes contain multiple G-rich elements. We show that efficient induction requires at least two copies of the CP1 element and that their relative orientation is unimportant. Two copies of an inverted relative orientation are, however, inactive when moved upstream of their normal position and are incapable of conferring cAMP-inducibility on a heterologous gene. These observations suggest that these sequences are either essential promoter elements, not themselves interacting with the inducer, or that their interaction with a separate class of control sequences is necessary for inducible expression.
Pears
C J
CJ
Imperial Cancer Research Fund, Clare Hall Laboratories, Herts, UK.
Williams
J G
JG
D001483:Base Sequence; D000242:Cyclic AMP / Q000494:pharmacology*; D003546:Cysteine Endopeptidases / Q000235:genetics*; D004262:DNA Restriction Enzymes; D004023:Dictyostelium / Q000201:enzymology / Q000235:genetics*; D005796:Genes / Q000187:drug effects*; D005800:Genes, Fungal / Q000187:drug effects*; D006147:Guanine; D008969:Molecular Sequence Data; D011401:Promoter Regions, Genetic; D012333:RNA, Messenger / Q000235:genetics; D014158:Transcription, Genetic
D012333:RNA, Messenger; D006147:Guanine; D000242:Cyclic AMP; D004262:DNA Restriction Enzymes; D003546:Cysteine Endopeptidases