Identification of a unique cAMP-response element in the gene encoding the cell adhesion molecule gp80 in Dictyostelium discoideum.
Extracellular cAMP serves as a chemoattractant as well as a signal which regulates gene expression during development of Dictyostelium discoideum. The cell adhesion molecule gp80 is expressed at the aggregation stage, between 6 and 10 h of development, and is known to be under cAMP regulation. Transcription of the gp80 gene is first turned on at a low, basal level at the preaggregation stage and is then greatly augmented by pulses of low levels of cAMP at the aggregation stage. Using cloned cDNA sequences, we have isolated genomic DNA fragments encompassing the gp80 gene. The gp80 gene has a single open reading frame, with multiple transcription start sites located downstream from a putative TATA box. Several short, repeated sequences in the upstream sequence have also been identified. The cloned 1.3-kilobase upstream DNA was sufficient to confer proper temporal and cAMP regulation on a gp80 minigene reporter in Dictyostelium cells. Deletional analysis of this 5'-flanking DNA led to the mapping of a cAMP-response element (CRE) in the gp80 gene to sequences between -306 and -289 base pairs upstream of the translational start site. Present within this region is a sequence we refer to as box 1 (TGGTGTG). The gp80 box 1-CRE binds specifically to a protein present in nuclear extracts, but binding is abolished when mutations are introduced in the box 1 sequence. The gp80 box 1-CRE shows little sequence homology to CREs of late developmental genes and the expression of gp80 may involve a distinct signal transduction pathway.
Desbarats
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Banting and Best Department of Medical Research, Charles H. Best Institute, University of Toronto, Ontario, Canada.
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Wong
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Siu
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D000595:Amino Acid Sequence; D000818:Animals; D001483:Base Sequence; D015815:Cell Adhesion Molecules / Q000235:genetics*; D003001:Cloning, Molecular; D000242:Cyclic AMP / Q000502:physiology*; D004252:DNA Mutational Analysis; D004023:Dictyostelium / Q000235:genetics*; D015966:Gene Expression Regulation, Fungal; D005800:Genes, Fungal; D008969:Molecular Sequence Data; D009838:Oligodeoxyribonucleotides / Q000737:chemistry; D011401:Promoter Regions, Genetic; D012045:Regulatory Sequences, Nucleic Acid; D012091:Repetitive Sequences, Nucleic Acid; D015183:Restriction Mapping; D012689:Sequence Homology, Nucleic Acid
D015815:Cell Adhesion Molecules; D009838:Oligodeoxyribonucleotides; D000242:Cyclic AMP