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.