Park Gyungsoon, Servin Jacqueline A, Turner Gloria E, Altamirano Lorena, Colot Hildur V, Collopy Patrick, Litvinkova Liubov, Li Liande, Jones Carol A, Diala Fitz-Gerald, Dunlap Jay C, Borkovich Katherine A
Department of Plant Pathology and Microbiology, Institute for Integrative Genome Biology, University of California, Riverside, CA 92521, USA.
Eukaryot Cell. 2011 Nov;10(11):1553-64. doi: 10.1128/EC.05140-11. Epub 2011 Sep 30.
Serine/threonine (S/T) protein kinases are crucial components of diverse signaling pathways in eukaryotes, including the model filamentous fungus Neurospora crassa. In order to assess the importance of S/T kinases to Neurospora biology, we embarked on a global analysis of 86 S/T kinase genes in Neurospora. We were able to isolate viable mutants for 77 of the 86 kinase genes. Of these, 57% exhibited at least one growth or developmental phenotype, with a relatively large fraction (40%) possessing a defect in more than one trait. S/T kinase knockouts were subjected to chemical screening using a panel of eight chemical treatments, with 25 mutants exhibiting sensitivity or resistance to at least one chemical. This brought the total percentage of S/T mutants with phenotypes in our study to 71%. Mutants lacking apg-1, an S/T kinase required for autophagy in other organisms, possessed the greatest number of phenotypes, with defects in asexual and sexual growth and development and in altered sensitivity to five chemical treatments. We showed that NCU02245/stk-19 is required for chemotropic interactions between female and male cells during mating. Finally, we demonstrated allelism between the S/T kinase gene NCU00406 and velvet (vel), encoding a p21-activated protein kinase (PAK) gene important for asexual and sexual growth and development in Neurospora.
丝氨酸/苏氨酸(S/T)蛋白激酶是真核生物中多种信号通路的关键组成部分,包括模式丝状真菌粗糙脉孢菌。为了评估S/T激酶对粗糙脉孢菌生物学的重要性,我们对粗糙脉孢菌中的86个S/T激酶基因进行了全面分析。我们能够分离出86个激酶基因中77个的可行突变体。其中,57%表现出至少一种生长或发育表型,相当一部分(40%)在不止一个性状上存在缺陷。对S/T激酶基因敲除突变体进行了一组八种化学处理的化学筛选,有25个突变体对至少一种化学物质表现出敏感性或抗性。这使得我们研究中具有表型的S/T突变体的总比例达到71%。缺乏apg-1(其他生物体自噬所需的一种S/T激酶)的突变体表现出的表型最多,在无性和有性生长发育以及对五种化学处理的敏感性改变方面存在缺陷。我们发现NCU02245/stk-19是交配过程中雌雄细胞之间趋化相互作用所必需的。最后,我们证明了S/T激酶基因NCU00406与velvet(vel)之间的等位关系,velvet编码一种对粗糙脉孢菌无性和有性生长发育重要的p21激活蛋白激酶(PAK)基因。