title: Temperature-sensitive inhibition of development in Dictyostelium due to a point mutation in the piaA gene.
abstract: The Dictyostelium mutant HSB1 is temperature-sensitive for development, undergoing aggregation and fruiting body formation at temperatures below 18 degrees C but not above. In vivo G protein-linked adenylyl cyclase activation is defective in HSB1, and the enzyme is not stimulated in vitro by GTPgammaS; stimulation is restored upon addition of wild-type cytosol. Transfection with the gene encoding the cytosolic regulator PIA rescued the mutant. We excluded the possibility that HSB1 cells fail to express PIA and show that the HSB1 piaA gene harbors a point mutation, resulting in the amino acid exchange G(917)D. Both wild-type and HSB1 cells were also transfected with the HSB1 piaA gene. The piaA(HSB1) gene product displayed a partial inhibitory effect on wild-type cell development. We hypothesize that PIA couples the heterotrimeric G protein to adenylyl cyclase via two binding sites, one of which is altered in a temperature-sensitive way by the HSB1 mutation. When overexpressed in the wild-type background, PIA(HSB1) competes with wild-type PIA via the nonmutated binding site, resulting in dominant-negative inhibition of development. Expression of GFP-fused PIA shows that PIA is homogeneously distributed in the cytoplasm of chemotactically moving cells.
authors.affiliation: Dipartimento di Scienze Cliniche e Biologiche, Università di Torino, Ospedale S. Luigi, 10043, Orbassano, Italy.
authors.forename: Barbara
authors.initials: B
authors.lastname: Pergolizzi
authors.forename: Barbara
authors.initials: B
authors.lastname: Peracino
authors.forename: James
authors.initials: J
authors.lastname: Silverman
authors.forename: Adriano
authors.initials: A
authors.lastname: Ceccarelli
authors.forename: Angelika
authors.initials: A
authors.lastname: Noegel
authors.forename: Peter
authors.initials: P
authors.lastname: Devreotes
authors.forename: Salvatore
authors.initials: S
authors.lastname: Bozzaro
mesh_terms: D000818:Animals; D002633:Chemotaxis / Q000235:genetics; D004023:Dictyostelium / Q000235:genetics* / Q000254:growth & development*; D018507:Gene Expression Regulation, Developmental / Q000502:physiology; D017354:Point Mutation; D015800:Protozoan Proteins / Q000235:genetics*; D015398:Signal Transduction / Q000235:genetics; D013696:Temperature
chemical_list: C109557:PiaA protein, Dictyostelium; D015800:Protozoan Proteins