title: Gib2, a novel Gbeta-like/RACK1 homolog, functions as a Gbeta subunit in cAMP signaling and is essential in Cryptococcus neoformans.
abstract: Canonical G proteins are heterotrimeric, consisting of alpha, beta, and gamma subunits. Despite multiple Galpha subunits functioning in fungi, only a single Gbeta subunit per species has been identified, suggesting that non-conventional G protein signaling exists in this diverse group of eukaryotic organisms. Using the Galpha subunit Gpa1 that functions in cAMP signaling as bait in a two-hybrid screen, we have identified a novel Gbeta-like/RACK1 protein homolog, Gib2, from the human pathogenic fungus Cryptococcus neoformans. Gib2 contains a seven WD-40 repeat motif and is predicted to form a seven-bladed beta propeller structure characteristic of beta transducins. Gib2 is also shown to interact, respectively, with two Ggamma subunit homologs, Gpg1 and Gpg2, similar to the conventional Gbeta subunit Gpb1. In contrast to Gpb1 whose overexpression promotes mating response, overproduction of Gib2 suppresses defects of gpa1 mutation in both melanization and capsule formation, the phenotypes regulated by cAMP signaling and associated with virulence. Furthermore, depletion of Gib2 by antisense suppression results in a severe growth defect, suggesting that Gib2 is essential. Finally, Gib2 is shown to also physically interact with a downstream target of Gpa1-cAMP signaling, Smg1, and the protein kinase C homolog Pkc1, indicating that Gib2 is also a multifunctional RACK1-like protein.
authors.affiliation: Research Institute for Children, Department of Pediatrics, Louisiana State University Health Sciences Center, 200 Henry Clay Avenue, New Orleans, LA 70118, USA.
authors.forename: Daniel A
authors.initials: DA
authors.lastname: Palmer
authors.forename: Jill K
authors.initials: JK
authors.lastname: Thompson
authors.forename: Lie
authors.initials: L
authors.lastname: Li
authors.forename: Ashton
authors.initials: A
authors.lastname: Prat
authors.forename: Ping
authors.initials: P
authors.lastname: Wang
mesh_terms: D000595:Amino Acid Sequence; D003455:Cryptococcus neoformans / Q000472:pathogenicity / Q000502:physiology*; D000242:Cyclic AMP / Q000378:metabolism*; D044387:GTP-Binding Protein beta Subunits; D020962:Heterotrimeric GTP-Binding Proteins / Q000737:chemistry* / Q000235:genetics*; D008958:Models, Molecular; D008969:Molecular Sequence Data; D010802:Phylogeny; D017433:Protein Structure, Secondary; D017386:Sequence Homology, Amino Acid; D015398:Signal Transduction; D020798:Two-Hybrid System Techniques
chemical_list: D044387:GTP-Binding Protein beta Subunits; D000242:Cyclic AMP; D020962:Heterotrimeric GTP-Binding Proteins