Identification of a novel type of cGMP phosphodiesterase that is defective in the chemotactic stmF mutants.
StmF mutants are chemotactic mutants that are defective in a cGMP phosphodiesterase (PDE) activity. We identified a novel gene, PdeD, that harbors two cyclic nucleotide-binding domains and a metallo-beta-lactamase homology domain. Similar to stmF mutants, pdeD-null mutants displayed extensively streaming aggregates, prolonged elevation of cGMP levels after chemotactic stimulation, and reduced cGMP-PDE activity. PdeD transcripts were lacking in stmF mutant NP377, indicating that this mutant carries a PdeD lesion. Expression of a PdeD-YFP fusion protein in pdeD-null cells restored the normal cGMP response and showed that PdeD resides in the cytosol. When purified by immunoprecipitation, the PdeD-YFP fusion protein displayed cGMP-PDE activity, which was retained in a truncated construct that contained only the metallo-beta-lactamase domain.
Meima
Marcel E
ME
School of Life Sciences, University of Dundee, Dundee DD1 5EH, United Kingdom.
Biondi
Ricardo M
RM
Schaap
Pauline
P
D015106:3',5'-Cyclic-GMP Phosphodiesterases / Q000737:chemistry / Q000235:genetics / Q000378:metabolism*; D000595:Amino Acid Sequence; D000818:Animals; D002633:Chemotaxis / Q000502:physiology*; D006152:Cyclic GMP / Q000378:metabolism; D004023:Dictyostelium / Q000201:enzymology* / Q000502:physiology; D004789:Enzyme Activation; D018507:Gene Expression Regulation, Developmental; D006801:Humans; D008969:Molecular Sequence Data; D009154:Mutation; D017434:Protein Structure, Tertiary; D011993:Recombinant Fusion Proteins / Q000235:genetics / Q000378:metabolism*; D016415:Sequence Alignment
D011993:Recombinant Fusion Proteins; D015106:3',5'-Cyclic-GMP Phosphodiesterases; D006152:Cyclic GMP