Structural Basis for the Interaction between the Potato Virus X Resistance Protein (Rx) and Its Cofactor Ran GTPase-activating Protein 2 (RanGAP2)

Hao, Wei; Collier, Sarah M.; Moffett, Peter; Chai, Jijie

doi:10.1074/jbc.m113.517417

Cited by 105 publications

(101 citation statements)

References 37 publications

Supporting

Mentioning

Contrasting

Order By: Relevance

“…The EDVID motif is conserved within this class of CC domains and is important in mediating intramolecular domain interactions in full-length CC-NLR proteins (26,28). In Sr33 6-120 , the equivalent motif (residues 77-81) encodes residues EDAVD, which reside in the α3 helix, with residues E77, D78, V80, and D81 all surface-exposed (Fig.…”

Section: Resultsmentioning

confidence: 99%

“…This result differed from our expectations based on the crystal structure of MLA10 . We investigated this finding further using in vitro biophysical techniques and also included the MLA10 5-120 and Rx 1-122 constructs used in previous structure determination studies (14,26), as well as Sr50 5-120 , a rye ortholog of Sr33 (4), in our analysis. Size-exclusion chromatography (SEC)-coupled multiangle light scattering (MALS) experiments on these four proteins found an average molecular mass in solution very close to the predicted monomer sizes, indicating that they were all predominantly monomeric in solution, even at loading concentrations of 30 mg/mL (Fig.…”

Section: E)mentioning

confidence: 99%

“…The N-terminal amino acids 5-120 of MLA10 (designated MLA10 ) crystallized as an antiparallel homodimer adopting a helix-loop-helix fold. However, the equivalent region from Rx (Rx 1-122 ) adopted a compact, fourhelical bundle when crystallized in a 1:1 heteroassociation with the Trp-Pro-Pro (WPP) domain from its cofactor protein RanGAP2 (26). MLA10 was also reported to dimerize in vitro, whereas Rx 1-122 was monomeric in vitro in the absence of cofactors.…”

mentioning

confidence: 99%

“…To date, structure-guided studies of plant NLRs have been restricted to the N-terminal TIR or CC domain (6,14,17,26), with the exception of the noncanonical integrated-sensor heavy metalassociated domain from the rice NLR Pik (27). Although the plant NLR TIR domains have a conserved fold (6,17), the structures of the two known CC domain fragments from barley MLA10 and potato Rx are strikingly different.…”

mentioning

confidence: 99%

See 3 more Smart Citations

The CC domain structure from the wheat stem rust resistance protein Sr33 challenges paradigms for dimerization in plant NLR proteins

Casey

Lavrencic

Bentham

et al. 2016

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

118

160

View full text Add to dashboard Cite

Plants use intracellular immunity receptors, known as nucleotidebinding oligomerization domain-like receptors (NLRs), to recognize specific pathogen effector proteins and induce immune responses. These proteins provide resistance to many of the world's most destructive plant pathogens, yet we have a limited understanding of the molecular mechanisms that lead to defense signaling. We examined the wheat NLR protein, Sr33, which is responsible for strainspecific resistance to the wheat stem rust pathogen, Puccinia graminis f. sp. tritici. We present the solution structure of a coiled-coil (CC) fragment from Sr33, which adopts a four-helix bundle conformation. Unexpectedly, this structure differs from the published dimeric crystal structure of the equivalent region from the orthologous barley powdery mildew resistance protein, MLA10, but is similar to the structure of the distantly related potato NLR protein, Rx. We demonstrate that these regions are, in fact, largely monomeric and adopt similar folds in solution in all three proteins, suggesting that the CC domains from plant NLRs adopt a conserved fold. However, larger C-terminal fragments of Sr33 and MLA10 can self-associate both in vitro and in planta, and this self-association correlates with their cell death signaling activity. The minimal region of the CC domain required for both cell death signaling and self-association extends to amino acid 142, thus including 22 residues absent from previous biochemical and structural protein studies. These data suggest that self-association of the minimal CC domain is necessary for signaling but is likely to involve a different structural basis than previously suggested by the MLA10 crystallographic dimer.plant innate immunity | resistance protein | NLR proteins | effectortriggered immunity | nuclear magnetic resonance spectroscopy

show abstract

Section: Resultsmentioning

confidence: 99%

Section: E)mentioning

confidence: 99%

mentioning

confidence: 99%

mentioning

confidence: 99%

See 2 more Smart Citations

The CC domain structure from the wheat stem rust resistance protein Sr33 challenges paradigms for dimerization in plant NLR proteins

Casey

Lavrencic

Bentham

et al. 2016

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

118

160

View full text Add to dashboard Cite

show abstract

“…However, as recent structural analyses have shown, EDVID-type CC domains can have significantly different tertiary and quaternary structures. The MLA10 CC domain forms homodimers and induces cell death (Maekawa et al, 2011), while the CC domain of Rx shows a tertiary structure that is markedly different from that of MLA10, does not form dimers, and does not induce cell death (Hao et al, 2013). As a result, signaling and/or selfinteraction may represent the exception, or the rule, or may simply reflect the different propensities of CC domains to function on their own.…”

Section: Discussion Activation Of Cell Death By the CC Domains Of I2-mentioning

confidence: 99%

The Chloroplastic Protein THF1 Interacts with the Coiled-Coil Domain of the Disease Resistance Protein N′ and Regulates Light-Dependent Cell Death

et al. 2016

Self Cite

View full text Add to dashboard Cite

One branch of plant immunity is mediated through nucleotide-binding/Leu-rich repeat (NB-LRR) family proteins that recognize specific effectors encoded by pathogens. Members of the I2-like family constitute a well-conserved subgroup of NB-LRRs from Solanaceae possessing a coiled-coil (CC) domain at their N termini. We show here that the CC domains of several I2-like proteins are able to induce a hypersensitive response (HR), a form of programmed cell death associated with disease resistance. Using yeast two-hybrid screens, we identified the chloroplastic protein Thylakoid Formation1 (THF1) as an interacting partner for several I2-like CC domains. Co-immunoprecipitations and bimolecular fluorescence complementation assays confirmed that THF1 and I2-like CC domains interact in planta and that these interactions take place in the cytosol. Several HR-inducing I2-like CC domains have a negative effect on the accumulation of THF1, suggesting that the latter is destabilized by active CC domains. To confirm this model, we investigated N9, which recognizes the coat protein of most Tobamoviruses, as a prototypical member of the I2-like family. Transient expression and gene silencing data indicated that THF1 functions as a negative regulator of cell death and that activation of full-length N9 results in the destabilization of THF1. Consistent with the known function of THF1 in maintaining chloroplast homeostasis, we show that the HR induced by N9 is light-dependent. Together, our results define, to our knowledge, novel molecular mechanisms linking light and chloroplasts to the induction of cell death by a subgroup of NB-LRR proteins.

show abstract

Plant Resistance to Infection by Viruses

Revers¹,

Nicaise²

2014

Encyclopedia of Life Sciences

View full text Add to dashboard Cite

Among the most devastating pathogens, plant viruses are responsible for considerable losses of most agronomically important crops in the fields. The development and use of cultivars that are genetically resistant to viruses is an efficient strategy to tackle the problems associated with virus diseases. Recently, important advances have been made in our understanding of the molecular nature and mechanisms associated with natural recessive and dominant resistance genes, as well as in the antiviral defence system based on ribonucleic acids, referred to as RNA silencing. In this review, the authors summarise current knowledge about the defence mechanisms that prevent or limit viral infection in plants and discuss the likely collaboration of these different antiviral strategies insuring the resistance of the whole plant. Key Concepts: A vast number of natural recessive resistances to plant viruses encode translational factors that are fully required for infection success. Most of plant antiviral R genes encode NBS‐LRR proteins that mediate resistance through the specific (direct or indirect) recognition of a virus avirulence factor. Plant hormone balance participates actively to plant virus resistance and can be modulated by viruses. The mechanisms of RNA silencing (or RNA interference) target virus multiplication at the nucleic acid level. Plant immunity against viruses results from a highly efficient build‐up of all resistance mechanisms.

show abstract

Structural Basis for the Interaction between the Potato Virus X Resistance Protein (Rx) and Its Cofactor Ran GTPase-activating Protein 2 (RanGAP2)

Cited by 105 publications

References 37 publications

The CC domain structure from the wheat stem rust resistance protein Sr33 challenges paradigms for dimerization in plant NLR proteins

The CC domain structure from the wheat stem rust resistance protein Sr33 challenges paradigms for dimerization in plant NLR proteins

The Chloroplastic Protein THF1 Interacts with the Coiled-Coil Domain of the Disease Resistance Protein N′ and Regulates Light-Dependent Cell Death

Plant Resistance to Infection by Viruses

Contact Info

Product

Resources

About