Germinal Center Kinases SmKIN3 and SmKIN24 Are Associated with the Sordaria macrospora Striatin-Interacting Phosphatase and Kinase (STRIPAK) Complex
Germinal Center Kinases SmKIN3 and SmKIN24 Are Associated with the Sordaria macrospora Striatin-Interacting Phosphatase and Kinase (STRIPAK) Complex
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DOI：
10.1371/journal.pone.0139163
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发表时间：
2015-09-29
期刊：
影响因子：
3.7
通讯作者：
Poeggeler, Stefanie
中科院分区：
文献类型：
--
作者：
Frey, Stefan;Reschka, Eva J.;Poeggeler, Stefanie
纹状体相互作用磷酸酶和激酶（STRIPAK）复合物由纹状体、蛋白磷酸酶PP 2A和调节动物和真菌发育的蛋白激酶组成。在丝状子囊菌大孢球孢菌中，它是子实体发育和细胞融合所必需的。在这里，我们报告的存在和功能的子囊菌中的PAK相关激酶。以哺乳动物生殖中心激酶（GCK）MST 4、STK 24、STK 25和MINK 1为查询序列，我们鉴定了两个可能的SmKIN 3和SmKIN 24同源物。大孢子虫。BLASTP搜索显示，这两种激酶是保守的丝状子囊菌。通过酵母双杂交（Y2 H）相互作用研究验证了纹状体蛋白同源物PRO 11与SmKIN 3和SmKIN 24的物理相互作用，并通过免疫共沉淀（co-IP）验证了SmKIN 3。在体内定位发现，这两种激酶都存在于隔膜和Smkin 3和Smkin 24的缺失导致异常的隔膜分布。而Smkin 3的缺失导致相邻隔膜之间的距离更大，气生菌丝增加，Smkin 24的缺失导致隔膜间距更近，不育。虽然表型不同，但两种激酶似乎独立地起作用，因为双敲除菌株Δ Smkin 3/Δ Smkin 24显示每个单缺失菌株的组合表型。
The striatin-interacting phosphatase and kinase (STRIPAK) complex is composed of striatin, protein phosphatase PP2A and protein kinases that regulate development in animals and fungi. In the filamentous ascomycete Sordaria macrospora, it is required for fruitingbody development and cell fusion. Here, we report on the presence and function of STRI-PAK-associated kinases in ascomycetes. Using the mammalian germinal center kinases (GCKs) MST4, STK24, STK25 and MINK1 as query, we identified the two putative homologs SmKIN3 and SmKIN24 in S. macrospora. A BLASTP search revealed that both kinases are conserved among filamentous ascomycetes. The physical interaction of the striatin homolog PRO11 with SmKIN3 and SmKIN24 were verified by yeast two-hybrid (Y2H) interaction studies and for SmKIN3 by co-Immunoprecipitation (co-IP). In vivo localization found that both kinases were present at the septa and deletion of both Smkin3 and Smkin24 led to abnormal septum distribution. While deletion of Smkin3 caused larger distances between adjacent septa and increased aerial hyphae, deletion of Smkin24 led to closer spacing of septa and to sterility. Although phenotypically distinct, both kinases appear to function independently because the double-knockout strain Delta Smkin3/Delta Smkin24 displayed the combined phenotypes of each single-deletion strain.