Distinct Roles of Non-Overlapping Surface Regions of the Coiled-Coil Domain in the Potato Immune Receptor Rx1

Erik J. Slootweg, Laurentiu N. Spiridon, Eliza C. Martin, Wladimir I.L. Tameling, Philip D. Townsend, Rikus Pomp, Jan Roosien, Olga Drawska, Octavina C.A. Sukarta, Arjen Schots, Jan Willem Borst, Matthieu H.A.J. Joosten, Jaap Bakker, Geert Smant, Martin J. Cann, Andrei-Jose Petrescu, Aska Goverse

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2 Citations (Scopus)

Abstract

The intracellular immune receptor Rx1 of potato (Solanum tuberosum), which confers effector-triggered immunity to Potato virus X, consists of a central nucleotide-binding domain (NB-ARC) flanked by a carboxyl-terminal leucine-rich repeat (LRR) domain and an amino-terminal coiled-coil (CC) domain. Rx1 activity is strictly regulated by interdomain interactions between the NB-ARC and LRR, but the contribution of the CC domain in regulating Rx1 activity or immune signaling is not fully understood. Therefore, we used a structure-informed approach to investigate the role of the CC domain in Rx1 functionality.
Targeted mutagenesis of CC surface residues revealed separate regions required for the intramolecular and intermolecular interaction of the CC with the NB-ARC-LRR and the cofactor Ran GTPase-activating protein2 (RanGAP2), respectively. None of the mutant Rx1 proteins was constitutively active, indicating that the CC does not contribute to the autoinhibition of Rx1 activity. Instead, the CC domain acted as a modulator of downstream responses involved in effector-triggered immunity. Systematic disruption of the hydrophobic interface between the four helices of the CC enabled the uncoupling of cell death and disease resistance responses. Moreover, a strong dominant negative effect on Rx1-mediated resistance and cell death was observed upon coexpression of the CC alone with full-length Rx1 protein, which depended on the RanGAP2-binding surface of the CC. Surprisingly, coexpression of the N-terminal half of the CC enhanced Rx1-mediated resistance, which further indicated that the CC functions as a scaffold for downstream components involved in the modulation of disease resistance or cell death signaling.
Original languageEnglish
Pages (from-to)1310-1331
JournalPlant Physiology
Volume178
Issue number3
DOIs
Publication statusPublished - Nov 2018

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AIDS-Related Complex
Solanum tuberosum
Leucine
leucine
cell death
Disease Resistance
Cell Death
GTP Phosphohydrolases
guanosinetriphosphatase
potatoes
disease resistance
Immunity
immunity
Potexvirus
Potato virus X
site-directed mutagenesis
Mutant Proteins
Mutagenesis
Nucleotides
proteins

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Slootweg, E. J., Spiridon, L. N., Martin, E. C., Tameling, W. I. L., Townsend, P. D., Pomp, R., ... Goverse, A. (2018). Distinct Roles of Non-Overlapping Surface Regions of the Coiled-Coil Domain in the Potato Immune Receptor Rx1. Plant Physiology, 178(3), 1310-1331. https://doi.org/10.1104/pp.18.00603
Slootweg, Erik J. ; Spiridon, Laurentiu N. ; Martin, Eliza C. ; Tameling, Wladimir I.L. ; Townsend, Philip D. ; Pomp, Rikus ; Roosien, Jan ; Drawska, Olga ; Sukarta, Octavina C.A. ; Schots, Arjen ; Borst, Jan Willem ; Joosten, Matthieu H.A.J. ; Bakker, Jaap ; Smant, Geert ; Cann, Martin J. ; Petrescu, Andrei-Jose ; Goverse, Aska. / Distinct Roles of Non-Overlapping Surface Regions of the Coiled-Coil Domain in the Potato Immune Receptor Rx1. In: Plant Physiology. 2018 ; Vol. 178, No. 3. pp. 1310-1331.
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title = "Distinct Roles of Non-Overlapping Surface Regions of the Coiled-Coil Domain in the Potato Immune Receptor Rx1",
abstract = "The intracellular immune receptor Rx1 of potato (Solanum tuberosum), which confers effector-triggered immunity to Potato virus X, consists of a central nucleotide-binding domain (NB-ARC) flanked by a carboxyl-terminal leucine-rich repeat (LRR) domain and an amino-terminal coiled-coil (CC) domain. Rx1 activity is strictly regulated by interdomain interactions between the NB-ARC and LRR, but the contribution of the CC domain in regulating Rx1 activity or immune signaling is not fully understood. Therefore, we used a structure-informed approach to investigate the role of the CC domain in Rx1 functionality.Targeted mutagenesis of CC surface residues revealed separate regions required for the intramolecular and intermolecular interaction of the CC with the NB-ARC-LRR and the cofactor Ran GTPase-activating protein2 (RanGAP2), respectively. None of the mutant Rx1 proteins was constitutively active, indicating that the CC does not contribute to the autoinhibition of Rx1 activity. Instead, the CC domain acted as a modulator of downstream responses involved in effector-triggered immunity. Systematic disruption of the hydrophobic interface between the four helices of the CC enabled the uncoupling of cell death and disease resistance responses. Moreover, a strong dominant negative effect on Rx1-mediated resistance and cell death was observed upon coexpression of the CC alone with full-length Rx1 protein, which depended on the RanGAP2-binding surface of the CC. Surprisingly, coexpression of the N-terminal half of the CC enhanced Rx1-mediated resistance, which further indicated that the CC functions as a scaffold for downstream components involved in the modulation of disease resistance or cell death signaling.",
author = "Slootweg, {Erik J.} and Spiridon, {Laurentiu N.} and Martin, {Eliza C.} and Tameling, {Wladimir I.L.} and Townsend, {Philip D.} and Rikus Pomp and Jan Roosien and Olga Drawska and Sukarta, {Octavina C.A.} and Arjen Schots and Borst, {Jan Willem} and Joosten, {Matthieu H.A.J.} and Jaap Bakker and Geert Smant and Cann, {Martin J.} and Andrei-Jose Petrescu and Aska Goverse",
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language = "English",
volume = "178",
pages = "1310--1331",
journal = "Plant Physiology",
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Slootweg, EJ, Spiridon, LN, Martin, EC, Tameling, WIL, Townsend, PD, Pomp, R, Roosien, J, Drawska, O, Sukarta, OCA, Schots, A, Borst, JW, Joosten, MHAJ, Bakker, J, Smant, G, Cann, MJ, Petrescu, A-J & Goverse, A 2018, 'Distinct Roles of Non-Overlapping Surface Regions of the Coiled-Coil Domain in the Potato Immune Receptor Rx1', Plant Physiology, vol. 178, no. 3, pp. 1310-1331. https://doi.org/10.1104/pp.18.00603

Distinct Roles of Non-Overlapping Surface Regions of the Coiled-Coil Domain in the Potato Immune Receptor Rx1. / Slootweg, Erik J.; Spiridon, Laurentiu N.; Martin, Eliza C.; Tameling, Wladimir I.L.; Townsend, Philip D.; Pomp, Rikus; Roosien, Jan; Drawska, Olga; Sukarta, Octavina C.A.; Schots, Arjen; Borst, Jan Willem; Joosten, Matthieu H.A.J.; Bakker, Jaap; Smant, Geert; Cann, Martin J.; Petrescu, Andrei-Jose; Goverse, Aska.

In: Plant Physiology, Vol. 178, No. 3, 11.2018, p. 1310-1331.

Research output: Contribution to journalArticleAcademicpeer-review

TY - JOUR

T1 - Distinct Roles of Non-Overlapping Surface Regions of the Coiled-Coil Domain in the Potato Immune Receptor Rx1

AU - Slootweg, Erik J.

AU - Spiridon, Laurentiu N.

AU - Martin, Eliza C.

AU - Tameling, Wladimir I.L.

AU - Townsend, Philip D.

AU - Pomp, Rikus

AU - Roosien, Jan

AU - Drawska, Olga

AU - Sukarta, Octavina C.A.

AU - Schots, Arjen

AU - Borst, Jan Willem

AU - Joosten, Matthieu H.A.J.

AU - Bakker, Jaap

AU - Smant, Geert

AU - Cann, Martin J.

AU - Petrescu, Andrei-Jose

AU - Goverse, Aska

PY - 2018/11

Y1 - 2018/11

N2 - The intracellular immune receptor Rx1 of potato (Solanum tuberosum), which confers effector-triggered immunity to Potato virus X, consists of a central nucleotide-binding domain (NB-ARC) flanked by a carboxyl-terminal leucine-rich repeat (LRR) domain and an amino-terminal coiled-coil (CC) domain. Rx1 activity is strictly regulated by interdomain interactions between the NB-ARC and LRR, but the contribution of the CC domain in regulating Rx1 activity or immune signaling is not fully understood. Therefore, we used a structure-informed approach to investigate the role of the CC domain in Rx1 functionality.Targeted mutagenesis of CC surface residues revealed separate regions required for the intramolecular and intermolecular interaction of the CC with the NB-ARC-LRR and the cofactor Ran GTPase-activating protein2 (RanGAP2), respectively. None of the mutant Rx1 proteins was constitutively active, indicating that the CC does not contribute to the autoinhibition of Rx1 activity. Instead, the CC domain acted as a modulator of downstream responses involved in effector-triggered immunity. Systematic disruption of the hydrophobic interface between the four helices of the CC enabled the uncoupling of cell death and disease resistance responses. Moreover, a strong dominant negative effect on Rx1-mediated resistance and cell death was observed upon coexpression of the CC alone with full-length Rx1 protein, which depended on the RanGAP2-binding surface of the CC. Surprisingly, coexpression of the N-terminal half of the CC enhanced Rx1-mediated resistance, which further indicated that the CC functions as a scaffold for downstream components involved in the modulation of disease resistance or cell death signaling.

AB - The intracellular immune receptor Rx1 of potato (Solanum tuberosum), which confers effector-triggered immunity to Potato virus X, consists of a central nucleotide-binding domain (NB-ARC) flanked by a carboxyl-terminal leucine-rich repeat (LRR) domain and an amino-terminal coiled-coil (CC) domain. Rx1 activity is strictly regulated by interdomain interactions between the NB-ARC and LRR, but the contribution of the CC domain in regulating Rx1 activity or immune signaling is not fully understood. Therefore, we used a structure-informed approach to investigate the role of the CC domain in Rx1 functionality.Targeted mutagenesis of CC surface residues revealed separate regions required for the intramolecular and intermolecular interaction of the CC with the NB-ARC-LRR and the cofactor Ran GTPase-activating protein2 (RanGAP2), respectively. None of the mutant Rx1 proteins was constitutively active, indicating that the CC does not contribute to the autoinhibition of Rx1 activity. Instead, the CC domain acted as a modulator of downstream responses involved in effector-triggered immunity. Systematic disruption of the hydrophobic interface between the four helices of the CC enabled the uncoupling of cell death and disease resistance responses. Moreover, a strong dominant negative effect on Rx1-mediated resistance and cell death was observed upon coexpression of the CC alone with full-length Rx1 protein, which depended on the RanGAP2-binding surface of the CC. Surprisingly, coexpression of the N-terminal half of the CC enhanced Rx1-mediated resistance, which further indicated that the CC functions as a scaffold for downstream components involved in the modulation of disease resistance or cell death signaling.

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DO - 10.1104/pp.18.00603

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Slootweg EJ, Spiridon LN, Martin EC, Tameling WIL, Townsend PD, Pomp R et al. Distinct Roles of Non-Overlapping Surface Regions of the Coiled-Coil Domain in the Potato Immune Receptor Rx1. Plant Physiology. 2018 Nov;178(3):1310-1331. https://doi.org/10.1104/pp.18.00603