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Suppression of fruit-body formation by constitutively active G-protein -subunits ScGP-A and ScGP-C in the homobasidiomycete Schizophyllum commune

National Agricultural Research Centre for Tohoku Region, National Agricultural Research Organization, Arai, Fukushima, 960-2156, Japan¹

Author for correspondence: Kenji Yamagishi. Tel: +81 24 593 5151. Fax: +81 24 593 2155. e-mail: yamagisi{at}affrc.go.jp

The heterotrimeric G-protein -subunit family plays multiple roles in eukaryotic cells, such as the regulation of growth and development, of pathogenicity and of the transmission of pheromone stimulation. In the homobasidiomycete Schizophyllum commune, some genes encoding heterotrimeric G-protein -subunits (SCGP1, SCGP2, ScGP-A, ScGP-B and ScGP-C) have been reported. In this study, constitutively active mutants of ScGP-A, ScGP-B and ScGP-C were generated by site-directed mutagenesis and introduced into the S. commune monokaryon strain to investigate the function of each gene. Northern analysis showed that the mutated genes were strongly expressed when compared with endogenous G-proteins in many clones. Upon macroscopic examination, some transformed clones expressing ScGP-A (Q207R) and ScGP-C (Q204R) mutant genes exhibited a slight suppression of aerial-hyphae formation in the monokaryon strain. In contrast to the slight suppression of aerial-hyphae formation in the monokaryon, most clones expressing mutated ScGP-A or ScGP-C genes failed to form fruit-bodies in the dikaryon strain. This observation indicated that ScGP-A and ScGP-C played a role in suppressing fruit-body formation in the dikaryon. Furthermore, these phenotypes were similar to the phenotype of the thn mutant in S. commune to some extent. Since the thn-1 gene encodes a putative regulator of the G-protein signalling protein (RGS), ScGP-A and ScGP-C might be targets of thn-1.

Keywords: site-directed mutagenesis

Abbreviations: GPD, glyceraldehyde-3-phosphate dehydrogenase; RGS, G-protein signalling protein; ScGP-A, Schizophyllum commune G-protein -subunit A; ScGP-B, S. commune G-protein -subunit B; ScGP-C, S. commune G-protein -subunit C

The GenBank accession numbers for the sequences reported in this paper are AB066503, AB051903 and AB051904.

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