Primary structure, expression and developmental regulation of a Dictyostelium calcineurin A homologue.
cDNA clones for the catalytic subunit of Ca2+/calmodulin(CaM)-dependent protein phosphatase (calcineurin A, protein phosphatase 2B) from Dictyostelium discoideum were isolated by functional screening of a lambda gt11 lysogen expression library with labeled Dictyostelium CaM. A complete cDNA of 2146 bp predicts a protein of 623 amino acids with homology to calcineurin A from other organisms and a similar molecular architecture. However, the Dictyostelium protein contains N-terminal and C-terminal extra domains causing a significantly higher molecular mass than found in any of its known counterparts. Recombinant Dictyostelium calcineurin A was purified from Escherichia coli cells and shown to display similar enzymatic properties as the enzyme from other sources. On Western blots specific antibodies against the protein recognized a band of approximately 80 kDa that migrated with an endogenous CaM-binding activity. Both the mRNA for calcineurin A and the protein are expressed during the growth phase. During early development the abundance of the protein is reduced and then increases to peak after 10 h of starvation, when tight aggregates have formed.
Dammann
H
H
Fakultät für Biologie, Universität Konstanz, Germany.
Hellstern
S
S
Husain
Q
Q
Mutzel
R
R
D000595:Amino Acid Sequence; D000818:Animals; D000906:Antibodies / Q000276:immunology; D001483:Base Sequence; D015153:Blotting, Western; D019703:Calcineurin; D002118:Calcium / Q000378:metabolism; D002147:Calmodulin / Q000378:metabolism; D002148:Calmodulin-Binding Proteins / Q000737:chemistry* / Q000235:genetics / Q000276:immunology / Q000378:metabolism*; D002846:Chromatography, Affinity; D003001:Cloning, Molecular; D018076:DNA, Complementary / Q000235:genetics; D004023:Dictyostelium / Q000201:enzymology* / Q000235:genetics / Q000254:growth & development; D004791:Enzyme Inhibitors / Q000494:pharmacology; D004926:Escherichia coli / Q000235:genetics; D018507:Gene Expression Regulation, Developmental; D007093:Imidazoles / Q000494:pharmacology; D008555:Melitten / Q000494:pharmacology; D008969:Molecular Sequence Data; D010749:Phosphoprotein Phosphatases / Q000737:chemistry* / Q000235:genetics / Q000276:immunology / Q000378:metabolism*; D011994:Recombinant Proteins / Q000737:chemistry / Q000378:metabolism; D016415:Sequence Alignment; D017421:Sequence Analysis
D000906:Antibodies; D002147:Calmodulin; D002148:Calmodulin-Binding Proteins; D018076:DNA, Complementary; D004791:Enzyme Inhibitors; D007093:Imidazoles; D011994:Recombinant Proteins; D008555:Melitten; C031938:calmidazolium; D019703:Calcineurin; D010749:Phosphoprotein Phosphatases; D002118:Calcium