title: NADPH oxidase homologs are required for normal cell differentiation and morphogenesis in Dictyostelium discoideum.
abstract: Membrane-associated NADPH oxidase complexes catalyse the production of the superoxide anion radical from oxygen and NADPH. In mammalian systems, NADPH oxidases form a family of at least seven isoforms that participate in host defence and signalling pathways. We report here the cloning and the characterisation of slime mould Dictyostelium discoideum homologs of the mammalian heme-containing subunit of flavocytochrome b (gp91(phox)) (NoxA, NoxB and NoxC), of the small subunit of flavocytochrome b (p22(phox)) and of the cytosolic factor p67(phox). Null-mutants of either noxA, noxB, noxC or p22(phox) show aberrant starvation-induced development and are unable to produce spores. The overexpression of NoxA(myc2) in noxA null strain restores spore formation. Remarkably, the gene alg-2B, coding for one of the two penta EF-hand proteins in Dictyostelium, acts as a suppressor in noxA, noxB, and p22(phox) null-mutant strains. Knockout of alg-2B allows noxA, noxB or p22(phox) null-mutants to return to normal development. However, the knockout of gene encoding NoxC, which contains two penta EF-hands, is not rescued by the invalidation of alg-2B. These data are consistent with a hypothesis connecting superoxide and calcium signalling during Dictyostelium development.
authors.affiliation: Laboratoire de Biochimie et Biophysique des Systèmes Intégrés (UMR5092 CNRS), Département de Réponse et Dynamique Cellulaires, CEA-Grenoble, France. BLardy@chu-grenoble.fr
authors.forename: Bernard
authors.initials: B
authors.lastname: Lardy
authors.forename: Mireille
authors.initials: M
authors.lastname: Bof
authors.forename: Laurence
authors.initials: L
authors.lastname: Aubry
authors.forename: Marie Hélène
authors.initials: MH
authors.lastname: Paclet
authors.forename: Françoise
authors.initials: F
authors.lastname: Morel
authors.forename: Michel
authors.initials: M
authors.lastname: Satre
authors.forename: Gérard
authors.initials: G
authors.lastname: Klein
mesh_terms: D000595:Amino Acid Sequence; D000818:Animals; D002384:Catalysis; D002454:Cell Differentiation; D002462:Cell Membrane / Q000201:enzymology; D002478:Cells, Cultured; D003001:Cloning, Molecular; D004023:Dictyostelium / Q000166:cytology / Q000201:enzymology* / Q000254:growth & development; D020832:EF Hand Motifs; D018507:Gene Expression Regulation, Developmental; D017125:Genes, Protozoan; D007527:Isoenzymes / Q000235:genetics / Q000378:metabolism; D008958:Models, Molecular; D008969:Molecular Sequence Data; D009024:Morphogenesis; D009154:Mutation; D019255:NADPH Oxidases / Q000737:chemistry / Q000235:genetics / Q000378:metabolism*; D010802:Phylogeny; D017433:Protein Structure, Secondary; D017434:Protein Structure, Tertiary; D017422:Sequence Analysis, DNA; D017386:Sequence Homology, Amino Acid; D013481:Superoxides / Q000378:metabolism
chemical_list: D007527:Isoenzymes; D013481:Superoxides; D019255:NADPH Oxidases