Cloning and expression in Escherichia coli of a cDNA encoding a developmentally regulated Ca(2+)-binding protein from Dictyostelium discoideum.
We have cloned a full-length cDNA from Dictyostelium discoideum which encodes a new Ca(2+)-binding protein. The deduced protein (termed CBP1) is composed of 156 amino acids and contains four consensus metal-ligating loop sequences found in helix-loop-helix motifs of many Ca(2+)-binding proteins. When expressed in bacteria as a GST fusion protein, CBP1 binds Ca2+ in a 45Ca2+ overlay assay. CBP1 exhibits little amino acid sequence homology with Dictyostelium calmodulin or calfumirin-1 (CAF-1) except in the putative Ca(2+)-binding regions. Moreover, unlike calmodulin and CAF-1 expression, CBP1 mRNA is expressed preferentially during the multicellular stages of development.
Coukell
B
B
Department of Biology, York University, Ont., Canada.
Moniakis
J
J
Grinberg
A
A
D000595:Amino Acid Sequence; D000818:Animals; D001483:Base Sequence; D002118:Calcium / Q000378:metabolism; D002135:Calcium-Binding Proteins / Q000737:chemistry / Q000235:genetics* / Q000378:metabolism; D003001:Cloning, Molecular; D018076:DNA, Complementary / Q000235:genetics*; D004023:Dictyostelium / Q000235:genetics* / Q000378:metabolism; D004926:Escherichia coli / Q000235:genetics; D018507:Gene Expression Regulation, Developmental; D017125:Genes, Protozoan / Q000235:genetics*; D018257:Helix-Loop-Helix Motifs / Q000235:genetics; D008969:Molecular Sequence Data; D015800:Protozoan Proteins; D012333:RNA, Messenger / Q000032:analysis; D016053:RNA, Protozoan / Q000032:analysis; D011993:Recombinant Fusion Proteins / Q000096:biosynthesis / Q000378:metabolism; D016415:Sequence Alignment; D017422:Sequence Analysis, DNA
D002135:Calcium-Binding Proteins; D018076:DNA, Complementary; D015800:Protozoan Proteins; D012333:RNA, Messenger; D016053:RNA, Protozoan; D011993:Recombinant Fusion Proteins; C093239:calcium-binding protein 1, Dictyostelium discoideum; D002118:Calcium