Schubert Daniela, Raudaskoski Marjatta, Knabe Nicole, Kothe Erika
Institute of Microbiology, Microbial Phytopathology, Neugasse 25, D-07743 Jena, Germany.
Eukaryot Cell. 2006 Apr;5(4):683-95. doi: 10.1128/EC.5.4.683-695.2006.
The white rot fungus Schizophyllum commune is used for the analysis of mating and sexual development in homobasidiomycete fungi. In this study, we isolated the gene gap1 encoding a GTPase-activating protein for Ras. Disruption of gap1 should therefore lead to strains accumulating Ras in its activated, GTP-bound state and to constitutive Ras signaling. Haploid Deltagap1 monokaryons of different mating types did not show alterations in mating behavior in the four different mating interactions possible in fungi expressing a tetrapolar mating type system. Instead, the growth rate in Deltagap1 monokaryons was reduced by ca. 25% and ca. 50% in homozygous Deltagap1/Deltagap1 dikaryons. Monokaryons, as well as homozygous dikaryons, carrying the disrupted gap1 alleles exhibited a disorientated growth pattern. Dikaryons showed a strong phenotype during clamp formation since hook cells failed to fuse with the peg beside them. Instead, the dikaryotic character of the hyphae was rescued by fusion of the hooks with nearby developing branches. Deltagap1/Deltagap1 dikaryons formed increased numbers of fruitbody primordia, whereas the amount of fruitbodies was not raised. Mature fruitbodies formed no or abnormal gills. No production of spores could be observed. The results suggest Ras involvement in growth, clamp formation, and fruitbody development.
白腐真菌裂褶菌用于分析同担子菌纲真菌的交配和有性发育。在本研究中,我们分离出了编码Ras的GTP酶激活蛋白的基因gap1。因此,gap1的破坏应导致菌株在其激活的、GTP结合状态下积累Ras,并导致Ras信号的组成型表达。在表达四极性交配型系统的真菌中可能存在的四种不同交配相互作用中,不同交配型的单倍体Deltagap1单核体在交配行为上未显示出变化。相反,Deltagap1单核体的生长速率降低了约25%,而纯合Deltagap1/Deltagap1双核体的生长速率降低了约50%。携带破坏的gap1等位基因的单核体以及纯合双核体表现出生长模式紊乱。双核体在锁状联合形成过程中表现出强烈的表型,因为钩状细胞未能与其旁边的钉状细胞融合。相反,钩状细胞与附近发育中的分支融合挽救了菌丝的双核特征。Deltagap1/Deltagap1双核体形成的子实体原基数量增加,而子实体的数量并未增加。成熟子实体没有形成菌褶或形成异常菌褶。未观察到孢子产生。结果表明Ras参与生长、锁状联合形成和子实体发育。