NLR immune receptor RB is differentially targeted by two homologous but functionally distinct effector proteins

Plant nucleotide-binding leucine-rich repeat(NLR)receptors mediate immune responses by directly or indirectly sensing pathogen-derived effectors.Despite significant advances in the understanding of NLR-mediated immunity,the mechanisms by which pathogens evolve to suppress NLR activation triggered by cognate effectors and gain virulence remain largely unknown.The agronomically important immune receptor RB recognizes the ubiquitous and highly conserved IPI-O RXLR family members(e.g.,IPI-O1)from Phytophthora infestans,and this process is suppressed by the rarely present and homologous effector IPIO4.Here,we report that self-association of RB via the coiled-coil(CC)domain is required for RB activation and is differentially affected by avirulence and virulence effectors.IPI-O1 moderately reduces the self-association of RB CC,potentially leading to changes in the conformation and equilibrium of RB,whereas IPIO4 dramatically impairs CC self-association to prevent RB activation.We also found that IPI-O1 associates with itself,whereas IPI-O4 does not.Notably,IPI-O4 interacts with IPI-O1 and disrupts its self-association,therefore probably blocking its avirulence function.Furthermore,IPI-O4 enhances the interaction between RB CC and IPI-O1,possibly sequestering RB and IPI-O1 and subsequently blocking their interactions with signaling components.Taken together,these findings considerably extend our understanding of the underlying mechanisms by which emerging virulent pathogens suppress the NLR-mediated recognition of cognate effectors.

Extracellular and cytoplasmic regions of LRIG1 play a negative role in EGFR activity: Findings of a radioligand-binding assay

Objective Leucine-rich repeats and immunoglobulin-like domains 1(LRIG1) is a newly identified human gene that inhibits the epidermal growth factor receptor(EGFR), which on combining with a ligand, can drive tumor growth. This study investigated the interaction between human LRIG1 and EGFR and attempted to delineate the functions of as well as the mechanisms used by the extracellular(ECD) and cytoplasmic(CPD) domains of the human LRIG1 protein to downregulate human EGFR signaling activity.Methods Two constructed chimeric eukaryotic expression vectors, pIRES2-EGFP-3XFLAG-LRIG1-ET and p3FLAG-LRIG1-TC, encoding the extracellular and transmembrane regions(LRIG1-ET) and the transmembrane and cytoplasmic regions(LRIG1-TC), respectively, and the plasmid p3XFLAG-CMV-9-LRIG1 encoding full-length LRIG1(LRIG1-FL) were transfected into the human glioma cell line U251 or primary astrocytoma cells by using liposomes. The number and affinity of cell surface EGFR on transfected cells was determined by ^(125)I-EGF binding assay. Results The dissociation constant(KD) values for EGFR were higher, and the maximum increase was observed in the cells transfected into LRIG1-ET(1.36 folds). The number of maximal binding sites(Bmax) of the receptors was decreased in all transfected cells; the maximum decrease was noted in the cells transfected into LRIG1-FL(40.05%).Conclusion Both the ECD and CPD of LRIG1 are important to negate EGFR signaling. The ECD may interfere with the binding between EGFR and its ligand and facilitate the functions of CPD. The CPD may, when brought in proximity to EGFR, enhance receptor degradation. These two mechanisms can contribute to the downregulation of EGFR-mediated signaling by LRIG1.

Wheat zinc finger protein TaZF interacts with both the powdery mildew AvrPm2 protein and the corresponding wheat Pm2a immune receptor

Plant defense responses to pathogens are induced after direct or indirect perception of effector proteins or their activity on host proteins.In fungal–plant interactions,relatively little is known about whether,in addi-tion to avirulence effectors and immune receptors,other proteins contribute to specific recognition.The nucleotide-binding leucine-rich repeat(NLR)immune receptor Pm2a in wheat recognizes the fungal pow-dery mildew effector AvrPm2.We found that the predicted wheat zincfinger TaZF interacts with both the fungal avirulence protein AvrPm2 and the wheat NLR Pm2a.We further demonstrated that the virulent AvrPm2-H2 variant does not interact with TaZF.TaZF silencing in wheat resulted in a reduction but not a loss of Pm2a-mediated powdery mildew resistance.Interaction studies showed that the leucine-rich repeat domain of Pm2a is the mediator of the interaction with TaZF.TaZF recruits both Pm2a and AvrPm2 from the cytosol to the nucleus,resulting in nuclear localization of Pm2a,TaZF,and AvrPm2 in wheat.We propose that TaZF acts as a facilitator of Pm2a-dependent AvrPm2 effector recognition.Ourfindings highlight the importance of identifying effector host targets for characterization of NLR-mediated effector recognition.

A Plant Immune Receptor Adopts a Two-Step Recognition Mechanism to Enhance Viral Effector Perception

Plant intracellular nucleotide binding leucine-rich repeat (NLR) immune receptors play critical roles in pathoge n surveillance. Most plant NLRs characterized so far were found to use a single domain/sensor to recognize pathogen effectors. Here we report that the Sw-5b NLR immune receptor uses two distinct domains to detect the viral movement protein NSm encoded by tospovirus. In addition to its leucine-rich repeat (LRR) domain that has been previously reported, the N-terminal Solanaceae domain (SD) of Sw- 5b also interacts with NSm and a conserved 21-amino-acid region of NSm (NSm^21). The specific interaction between Sw-5b SD and NSm is required for releasing the inhibitory effect of coiled-coil domain on the NBARC- LRR region. Furthermore, we found that the binding of NSm affects the nucleotide binding activity of the NB-ARC-LRR in vitro, while Sw-5b NB-ARC-LRR is activated only when NSm and NSm^21 levels are high. Interestingly, Sw-5b SD could significantly enhanee the ability of the NB-ARC-LRR to detect low levels of NSm effector and facilitate its activation and induction of defense response. An Sw-5b SD mutant that is disrupted in NSm recognition failed to enhance the ability of the NB-ARC-LRR to sense low levels of NSm and NSm^21 . Taken together, our results suggest that Sw-5b SD functions as an extra sensor and the NB-ARC-LRR as an activator, and that Sw-5b NLR adopts a two-step recog nition mechanism to enhance viral effector perception.

两类免疫受体强强联手筑牢植物免疫防线

植物先天免疫系统在抵御病原菌入侵过程中发挥至关重要的作用,主要包括两个层次,即病原菌相关分子模式和效应因子分别触发的PTI和ETI免疫反应。PTI和ETI分别由植物细胞膜表面模式识别受体(PRRs)和胞内免疫受体(NLRs)激活,具有特异的激活机制,但是两者激活的下游免疫事件相互重叠。PTI和ETI是否为泾渭分明的两道防线,以及ETI与PTI下游事件为何如此相似,一直是植物免疫领域最受关注的问题之一。最近,中国科学院分子植物科学卓越创新中心辛秀芳团队与合作者利用拟南芥(Arabidopsis thaliana)与丁香假单胞杆菌(Pseudomonas syringae)互作系统对PTI和ETI在机制上的联系进行了研究。他们发现PRRs和共受体参与ETI,而活性氧的产生是联系PRRs和NLRs所介导的免疫早期信号事件。他们还发现NLRs信号能够迅速增强PTI关键因子的转录和蛋白水平,PTI的增强在ETI免疫反应中不可或缺。该研究从机制上解析了植物免疫领域中长期悬而未决的PTI与ETI相似性之谜,是该领域的一项突破性进展,为未来作物分子设计育种提供了新的启示。