Laboratory of Phytopathology, Wageningen University, Droevendaalsesteeg 1, 6708 PB, Wageningen, The Netherlands.
Mol Microbiol. 2013 Apr;88(2):352-70. doi: 10.1111/mmi.12190. Epub 2013 Mar 11.
For dispersal and host infection plant pathogens largely depend on asexual spores. Pathogenesis and sporulation are complex processes that are governed by cellular signalling networks including G-protein and phospholipid signalling. Oomycetes possess a family of novel proteins called GPCR-PIPKs (GKs) that are composed of a seven-transmembrane spanning (7-TM) domain fused to a phosphatidylinositol phosphate kinase (PIPK) domain. Based on this domain structure GKs are anticipated to link G-protein and phospholipid signal pathways; however, their functions are currently unknown. Expression analyses of the 12 GK genes in Phytophthora infestans and their orthologues in Phytophthora sojae, revealed differential expression during asexual development. PiGK1 and PiGK4 were fused to monomeric red fluorescent protein (mRFP) and ectopically expressed in P. infestans. In growing hyphae different subcellular distribution patterns were observed indicating that these two GKs act independently during development. We focused on the functional analyses of PiGK4. Its localization suggested involvement in cell differentiation and elongation and its 7-TM domain showed a canonical GPCR membrane topology. Silencing of GK4 and overexpression of full-length and truncated constructs in P. infestans revealed that PiGK4 is not only involved in spore germination and hyphal elongation but also in sporangia cleavage and infection.
对于传播和宿主感染,植物病原体在很大程度上依赖于无性孢子。发病机制和孢子形成是复杂的过程,受包括 G 蛋白和磷脂信号在内的细胞信号网络调控。卵菌门拥有一类称为 GPCR-PIPKs(GKs)的新型蛋白,它们由一个七跨膜(7-TM)结构域融合到一个磷酸肌醇磷酸激酶(PIPK)结构域组成。基于这个结构域,GKs 预计能够连接 G 蛋白和磷脂信号通路;然而,它们的功能目前尚不清楚。在马铃薯晚疫病菌和大豆疫霉菌中,对 12 个 GK 基因及其同源基因的表达分析表明,它们在无性发育过程中存在差异表达。将 PiGK1 和 PiGK4 融合到单体红色荧光蛋白(mRFP)中,并在马铃薯晚疫病菌中异位表达。在生长的菌丝中观察到不同的亚细胞分布模式,表明这两个 GKs 在发育过程中独立发挥作用。我们专注于 PiGK4 的功能分析。其定位表明它参与细胞分化和伸长,其 7-TM 结构域显示出典型的 GPCR 膜拓扑结构。在马铃薯晚疫病菌中沉默 GK4 并过表达全长和截短构建体表明,PiGK4 不仅参与孢子萌发和菌丝伸长,还参与游动孢子分裂和侵染。