RanGAP2 Mediates Nucleocytoplasmic Partitioning of the NB-LRR Immune Receptor Rx in the Solanaceae, Thereby Dictating Rx Function

Tameling, W.I.L.; Nooijen, Claudia; Ludwig, Nora R.; Botër, Marta; Slootweg, E.J.; Goverse, A.; Shirasu, Ken; Joosten, M.H.A.J.

doi:10.1105/tpc.110.077461

Cited by 141 publications

(162 citation statements)

References 101 publications

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“…In terms of compartment-dependent activation, Sr33 is more similar to the potato CC-NLR Rx, a nucleo-cytoplasmic protein that recognizes the potato virus X (PVX) coat protein (CP) (32,49). Rx interacts with the small cytosolic GTPase RanGAP2, which is required for Rx function (50,51) and affects Rx nucleocytoplasmic distribution (49).…”

Section: Discussionmentioning

confidence: 99%

Cytosolic activation of cell death and stem rust resistance by cereal MLA-family CC–NLR proteins

Césari¹,

Moore²,

Chen³

et al. 2016

Proc. Natl. Acad. Sci. U.S.A.

105

120

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Plants possess intracellular immune receptors designated “nucleotide-binding domain and leucine-rich repeat” (NLR) proteins that translate pathogen-specific recognition into disease-resistance signaling. The wheat immune receptors Sr33 and Sr50 belong to the class of coiled-coil (CC) NLRs. They confer resistance against a broad spectrum of field isolates of Puccinia graminis f. sp. tritici, including the Ug99 lineage, and are homologs of the barley powdery mildew-resistance protein MLA10. Here, we show that, similarly to MLA10, the Sr33 and Sr50 CC domains are sufficient to induce cell death in Nicotiana benthamiana. Autoactive CC domains and full-length Sr33 and Sr50 proteins self-associate in planta. In contrast, truncated CC domains equivalent in size to an MLA10 fragment for which a crystal structure was previously determined fail to induce cell death and do not self-associate. Mutations in the truncated region also abolish self-association and cell-death signaling. Analysis of Sr33 and Sr50 CC domains fused to YFP and either nuclear localization or nuclear export signals in N. benthamiana showed that cell-death induction occurs in the cytosol. In stable transgenic wheat plants, full-length Sr33 proteins targeted to the cytosol provided rust resistance, whereas nuclear-targeted Sr33 was not functional. These data are consistent with CC-mediated induction of both cell-death signaling and stem rust resistance in the cytosolic compartment, whereas previous research had suggested that MLA10-mediated cell-death and disease resistance signaling occur independently, in the cytosol and nucleus, respectively.

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Section: Discussionmentioning

confidence: 99%

Cytosolic activation of cell death and stem rust resistance by cereal MLA-family CC–NLR proteins

Césari¹,

Moore²,

Chen³

et al. 2016

Proc. Natl. Acad. Sci. U.S.A.

105

120

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“…This system has been widely used for the functional analysis of HR mediated by NLRs from both monocots and dicots (Leister et al, 2005;van Ooijen et al, 2008;Slootweg et al, 2010;Tameling et al, 2010;Bai et al, 2012;Qi et al, 2012). We have recently shown that this is an appropriate system for the functional analysis of HR modulated by Rp1-D21 and its regulators (Wang and Balint-Kurti, 2015;Wang et al, 2015cWang et al, , 2015d.…”

Section: Ccoaomt2mentioning

confidence: 99%

Maize Homologs of CCoAOMT and HCT, Two Key Enzymes in Lignin Biosynthesis, Form Complexes with the NLR Rp1 Protein to Modulate the Defense Response

Wang

Balint‐Kurti²

2016

Plant Physiol.

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Disease resistance (R) genes encode nucleotide binding Leu-rich-repeat (NLR) proteins that confer resistance to specific pathogens. Upon pathogen recognition they trigger a defense response that usually includes a so-called hypersensitive response (HR), a rapid localized cell death at the site of pathogen infection. Intragenic recombination between two maize (Zea mays) NLRs, Rp1-D and Rp1-dp2, resulted in the formation of a hybrid NLR, Rp1-D21, which confers an autoactive HR in the absence of pathogen infection. From a previous quantitative trait loci and genome-wide association study, we identified genes encoding two key enzymes in lignin biosynthesis, hydroxycinnamoyltransferase (HCT) and caffeoyl CoA O-methyltransferase (CCoAOMT), adjacent to the nucleotide polymorphisms that were highly associated with variation in the severity of Rp1-D21-induced HR. We have previously shown that the two maize HCT homologs suppress the HR conferred by Rp1-D21 in a heterologous system, very likely through physical interaction. Here, we show, similarly, that CCoAOMT2 suppresses the HR induced by either the full-length or by the N-terminal coiled-coil domain of Rp1-D21 also likely via physical interaction and that the metabolic activity of CCoAOMT2 is unlikely to be necessary for its role in suppressing HR. We also demonstrate that CCoAOMT2, HCTs, and Rp1 proteins can form in the same complexes. A model is derived to explain the roles of CCoAOMT and HCT in Rp1-mediated defense resistance.

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“…However, no evidence exists that RanGAPs themselves can shuttle proteins from the cytoplasm to the nucleus; therefore, it is not likely that RanGAP2 has a direct role in the transport of Rx1 to the nucleus. On the other hand, a detailed study on the effect RanGAP2 has on the subcellular localization and activation of Rx1 (Tameling et al, 2010) showed that RanGAP2 affects the nucleocytoplasmic partitioning of Rx1 but does not require GAP (GTPase activating protein) activity to produce this effect.…”

Section: The CC and Lrr Domains Play Distinct Roles In The Nucleocytomentioning

confidence: 99%

“…If this is true, and a similar indirect RanGAP2-mediated recognition underlies PVX CP recognition, then this would explain why the nuclear-targeted PVX CP no longer activates Rx1. However, a study by Tameling et al (2010) determining the link between RanGAP2 localization and Rx1 functioning demonstrated that when the RanGAP2 WPP domain itself is targeted to the nucleus by an NLS fusion, thereby pulling Rx1 into the nucleus as well, it still does not allow Rx1 activation in the nuclear compartment. However, it might be the full-length RanGAP2 that is required for elicitor recognition.…”

Section: Rx1 Is Activated In the Cytoplasmic Compartmentmentioning

confidence: 99%

Nucleocytoplasmic Distribution Is Required for Activation of Resistance by the Potato NB-LRR Receptor Rx1 and Is Balanced by Its Functional Domains

Slootweg¹,

Roosien²,

et al. 2010

Self Cite

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The Rx1 protein, as many resistance proteins of the nucleotide binding-leucine-rich repeat (NB-LRR) class, is predicted to be cytoplasmic because it lacks discernable nuclear targeting signals. Here, we demonstrate that Rx1, which confers extreme resistance to Potato virus X, is located both in the nucleus and cytoplasm. Manipulating the nucleocytoplasmic distribution of Rx1 or its elicitor revealed that Rx1 is activated in the cytoplasm and cannot be activated in the nucleus. The coiled coil (CC) domain was found to be required for accumulation of Rx1 in the nucleus, whereas the LRR domain promoted the localization in the cytoplasm. Analyses of structural subdomains of the CC domain revealed no autonomous signals responsible for active nuclear import. Fluorescence recovery after photobleaching and nuclear fractionation indicated that the CC domain binds transiently to large complexes in the nucleus. Disruption of the Rx1 resistance function and protein conformation by mutating the ATP binding phosphate binding loop in the NB domain, or by silencing the cochaperone SGT1, impaired the accumulation of Rx1 protein in the nucleus, while Rx1 versions lacking the LRR domain were not affected in this respect. Our results support a model in which interdomain interactions and folding states determine the nucleocytoplasmic distribution of Rx1.

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RanGAP2 Mediates Nucleocytoplasmic Partitioning of the NB-LRR Immune Receptor Rx in the Solanaceae, Thereby Dictating Rx Function

Cited by 141 publications

References 101 publications

Cytosolic activation of cell death and stem rust resistance by cereal MLA-family CC–NLR proteins

Cytosolic activation of cell death and stem rust resistance by cereal MLA-family CC–NLR proteins

Maize Homologs of CCoAOMT and HCT, Two Key Enzymes in Lignin Biosynthesis, Form Complexes with the NLR Rp1 Protein to Modulate the Defense Response

Nucleocytoplasmic Distribution Is Required for Activation of Resistance by the Potato NB-LRR Receptor Rx1 and Is Balanced by Its Functional Domains

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