The Sainsbury Laboratory, Norwich Research Park, Norwich, United Kingdom.
PLoS Pathog. 2012;8(8):e1002875. doi: 10.1371/journal.ppat.1002875. Epub 2012 Aug 23.
Phytopathogenic oomycetes, such as Phytophthora infestans, secrete an arsenal of effector proteins that modulate plant innate immunity to enable infection. We describe CRN8, a host-translocated effector of P. infestans that has kinase activity in planta. CRN8 is a modular protein of the CRN effector family. The C-terminus of CRN8 localizes to the host nucleus and triggers cell death when the protein is expressed in planta. Cell death induction by CRN8 is dependent on its localization to the plant nucleus, which requires a functional nuclear localization signal (NLS). The C-terminal sequence of CRN8 has similarity to a serine/threonine RD kinase domain. We demonstrated that CRN8 is a functional RD kinase and that its auto-phosphorylation is dependent on an intact catalytic site. Co-immunoprecipitation experiments revealed that CRN8 forms a dimer or multimer. Heterologous expression of CRN8 in planta resulted in enhanced virulence by P. infestans. In contrast, in planta expression of the dominant-negative CRN8(R469A;D470A) resulted in reduced P. infestans infection, further implicating CRN8 in virulence. Overall, our results indicate that similar to animal parasites, plant pathogens also translocate biochemically active kinase effectors inside host cells.
植物病原卵菌,如致病疫霉,会分泌大量效应蛋白来调节植物固有免疫,从而实现侵染。我们描述了 P. infestans 的一个宿主易位效应子 CRN8,其在植物体内具有激酶活性。CRN8 是 CRN 效应家族的一种模块化蛋白。CRN8 的 C 端定位于宿主细胞核,当该蛋白在植物体内表达时会引发细胞死亡。CRN8 诱导的细胞死亡依赖于其在植物细胞核内的定位,这需要一个功能正常的核定位信号(NLS)。CRN8 的 C 端序列与丝氨酸/苏氨酸 RD 激酶结构域具有相似性。我们证明 CRN8 是一种功能性 RD 激酶,其自动磷酸化依赖于完整的催化位点。共免疫沉淀实验表明 CRN8 形成二聚体或多聚体。CRN8 在植物体内的异源表达导致 P. infestans 的毒力增强。相比之下,在植物体内表达显性失活的 CRN8(R469A;D470A)会导致 P. infestans 感染减少,这进一步表明 CRN8 与毒力有关。总的来说,我们的结果表明,与动物寄生虫类似,植物病原体也会将具有生物化学活性的激酶效应子易位到宿主细胞内。