Guanylate cyclase in Dictyostelium discoideum with the topology of mammalian adenylate cyclase.
The core of adenylate and guanylate cyclases is formed by an intramolecular or intermolecular dimer of two cyclase domains arranged in an antiparallel fashion. Metazoan membrane-bound adenylate cyclases are composed of 12 transmembrane spanning regions, and two cyclase domains which function as a heterodimer and are activated by G-proteins. In contrast, membrane-bound guanylate cyclases have only one transmembrane spanning region and one cyclase domain, and are activated by extracellular ligands to form a homodimer. In the cellular slime mould, Dictyostelium discoideum, membrane-bound guanylate cyclase activity is induced after cAMP stimulation; a G-protein-coupled cAMP receptor and G-proteins are essential for this activation. We have cloned a Dictyostelium gene, DdGCA, encoding a protein with 12 transmembrane spanning regions and two cyclase domains. Sequence alignment demonstrates that the two cyclase domains are transposed, relative to these domains in adenylate cyclases. DdGCA expressed in Dictyostelium exhibits high guanylate cyclase activity and no detectable adenylate cyclase activity. Deletion of the gene indicates that DdGCA is not essential for chemotaxis or osmo-regulation. The knock-out strain still exhibits substantial guanylate cyclase activity, demonstrating that Dictyostelium contains at least one other guanylate cyclase.
Department of Biochemistry, University of Groningen, Nijenborgh 4, 9747AG Groningen, The Netherlands.
J
J
Roelofs
H
H
Snippe
R G
RG
Kleineidam
P J
PJ
Van Haastert
D000262:Adenylyl Cyclases / Q000737:chemistry*; D000595:Amino Acid Sequence; D000818:Animals; D002449:Cell Aggregation; D002478:Cells, Cultured; D003001:Cloning, Molecular; D006152:Cyclic GMP / Q000378:metabolism; D004023:Dictyostelium / Q000166:cytology / Q000201:enzymology* / Q000235:genetics; D017353:Gene Deletion; D006162:Guanylate Cyclase / Q000737:chemistry* / Q000235:genetics* / Q000378:metabolism / Q000502:physiology; D008565:Membrane Proteins / Q000737:chemistry* / Q000235:genetics* / Q000502:physiology; D008969:Molecular Sequence Data; D010802:Phylogeny; D011487:Protein Conformation; D017434:Protein Structure, Tertiary; D015800:Protozoan Proteins; D017386:Sequence Homology, Amino Acid
D008565:Membrane Proteins; D015800:Protozoan Proteins; C424686:DdGCA protein, Dictyostelium discoideum; D000262:Adenylyl Cyclases; D006162:Guanylate Cyclase; D006152:Cyclic GMP