Mutagenesis of putative acylation sites alters function, localization, and accumulation of a Gi alpha subunit of the chestnut blight fungus Cryphonectria parasitica.
Targeted disruption of cpg-1, a gene encoding the G protein Gi alpha subunit, CPG-1, in the chestnut blight fungus, Cryphonectria parasitica, results in reduced mycelial growth, reduced orange pigmentation, loss of virulence, loss of asexual sporulation, and female infertility. We report the development of a complementation system for cpg-1 null mutants and its use to evaluate the in vivo consequences of mutating conserved putative CPG-1 myristoylation (G2) and palmitoylation (C3) sites. Independent mutations of the two putative acylation sites differentially altered complex fungal biological processes, including virulence, and modified CPG-1 membrane association. Results of combined Northern (RNA) and Western (immunoblot) analysis also indicated a role for lipid modification in post-transcriptional regulation of CPG-1 accumulation.
Gao
S
S
Center for Agricultural Biotechnology, University of Maryland Biotechnology Institute, University of Maryland, College Park 20742-3351, USA.
Nuss
D L
DL
D000215:Acylation; D001203:Ascomycota / Q000378:metabolism* / Q000502:physiology; D001483:Base Sequence; D017931:DNA Primers; D019206:GTP-Binding Protein alpha Subunits, Gi-Go / Q000378:metabolism*; D005816:Genetic Complementation Test; D016297:Mutagenesis, Site-Directed; D010641:Phenotype; D015398:Signal Transduction
D017931:DNA Primers; D019206:GTP-Binding Protein alpha Subunits, Gi-Go