The Irish potato famine pathogen Phytophthora infestans translocates the CRN8 kinase into host plant cells.
The Irish potato famine pathogen Phytophthora infestans translocates the CRN8 kinase into host plant cells.
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DOI：
10.1371/journal.ppat.1002875
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发表时间：
2012
期刊：
影响因子：
6.7
通讯作者：
Kamoun S
中科院分区：
文献类型：
--
作者：
van Damme M;Bozkurt TO;Cakir C;Schornack S;Sklenar J;Jones AM;Kamoun S
植物病原性卵菌，如致病疫霉，分泌大量的效应蛋白，调节植物的先天免疫，使其能够感染。我们描述了CRN8，一个寄主易位的致病假单胞菌的效应器，它在植物中具有激酶活性。CRN8是CRN效应器家族的一种模块化蛋白。当CRN8蛋白在植物中表达时，CRN8的C末端定位于宿主核并触发细胞死亡。CRN8诱导的细胞死亡依赖于其在植物核中的定位，这需要一个功能性的核定位信号(NLS)。CRN8的C末端序列与丝氨酸/苏氨酸RD激活区相似。我们证明了CRN8是一个功能性的RD激酶，它的自动磷酸化依赖于一个完整的催化位点。免疫共沉淀实验表明CRN8形成二聚体或多聚体。CRN8在植物中的异源表达可增强致病疫霉的致病力。相反，在植物中表达显性阴性的CRN8R469A；D470A导致致病疫霉感染减少，进一步暗示CRN8的毒力。总体而言，我们的结果表明，与动物寄生虫类似，植物病原体也将生物化学活性激酶效应器转移到宿主细胞内。马铃薯晚疫病菌是马铃薯和番茄晚疫病的病原菌。现在公认的是，卵菌，如致病疫霉，会分泌大量的效应蛋白，调节植物的先天免疫，从而使感染成为可能。含有LFLAK易位基序的Crinkler(CRN)效应器家族代表了一类来自卵菌的新的细胞质效应器。N-末端结构域包括这个基序，对于植物细胞死亡的诱导是必不可少的，这表明存在参与不同过程的模块结构域。CRN8的C末端/效应域定位于宿主核，核定位是触发细胞死亡所必需的。CRN8的效应域与植物丝氨酸/苏氨酸RD激酶有相似之处。这一有趣的发现提出了CRN8模仿一类特定植物酶的可能性，这一特征已经在其他几种病原体效应器中观察到。通过生化分析，我们证实了CRN8的C-末端结构域具有激酶活性，并通过质谱学鉴定了5个被证明是自动磷酸化的丝氨酸。据我们所知，CRN8是第一个报道的来自微生物植物病原体的分泌型活性激酶。
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 CRN8R469A;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. Phytophthora infestans is the causal agent of late blight on potato and tomato. It is now well established that oomycetes, such as P. infestans, secrete an arsenal of effector proteins that modulate plant innate immunity to enable infection. The Crinkler (CRN) effector family containing the LFLAK translocation motif represents a new class of cytoplasmic effectors from oomycetes. The N-terminal domain, which includes this motif, is dispensable for cell death induction in planta, indicating the presence of modular domains that are involved in distinct processes. The C-terminal/effector domain of CRN8 localizes to the host nucleus and nuclear localization is required for triggering cell death. The CRN8 effector domain has similarity to plant Serine/Threonine RD kinases. This intriguing finding raises the possibility that CRN8 mimics a specific class of plant enzymes, a feature that has been observed for several other pathogen effectors. Using biochemical assays, we confirmed that the C-terminal domain of CRN8 has kinase activity, and have identified by mass spectrometry five serines shown to be auto-phosphorylated. To the best of our knowledge, CRN8 is the first reported secreted active kinase from a microbial plant pathogen.
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影响因子：
11.6
作者：
Dou, Daolong;Kale, Shiv D.;Tyler, Brett M.
通讯作者：
Tyler, Brett M.
影响因子：
4.8
作者：
Dukuzumuremyi, JM;Rosqvist, R;Schesser, K
通讯作者：
Schesser, K
DOI：
10.1073/pnas.1112708109
发表时间：
2011-12-20
影响因子：
11.1
作者：
Bozkurt, Tolga O.;Schornack, Sebastian;Kamoun, Sophien
通讯作者：
Kamoun, Sophien
影响因子：
56.9
作者：
Allen, RL;Bittner-Eddy, PD;Beynon, JL
通讯作者：
Beynon, JL
影响因子：
7.2
作者：
Bos, Jorunn I. B.;Kanneganti, Thirumala-Devi;Kamoun, Sophien
通讯作者：
Kamoun, Sophien