The Potato Nucleotide-Binding Leucine-Rich Repeat (NLR) Immune Receptor Rx1 is a Pathogen Dependent DNA-Deforming Protein

Fenyk, Stepan; Townsend, Philip D.; Dixon, Christopher H.; Spies, Gerhard B.; de San Eustaquio Campillo, Alba; Slootweg, Erik J.; Westerhof, Lotte B.; Gawehns, Fleur K.K.; Knight, Marc R.; Sharples, Gary J.; Goverse, Aska; Pålsson, Lars-Olof; Takken, Frank L.W.; Cann, Martin J.

Abstract

Plant NLR proteins enable cells to respond to pathogen attack. Several NLRs act in the nucleus, however, conserved nuclear targets that support their role in immunity are unknown. Previously we noted a structural homology between the NB domain of NLRs and DNA replication origin-binding Cdc6/Orc1 proteins. Here we show that the NB-ARC domain of the Rx1 NLR of potato binds nucleic acids. Rx1 induces ATP-dependent bending and melting of DNA in vitro dependent upon a functional P-loop. In situ full-length Rx1 binds nuclear DNA following activation by its cognate pathogen-derived effector protein, the coat protein of potato virus X. In line with its obligatory nucleocytoplasmic distribution, DNA-binding was only observed when Rx1 was allowed to freely translocate between both compartments and was activated in the cytoplasm. Immune activation induced by an unrelated NLR-effector pair did not trigger a Rx1-DNA interaction. DNA-binding is therefore not merely a consequence of immune activation. These data establish a role for DNA distortion in Rx1 immune signalling and defines DNA as a molecular target of an activated NLR.

Citation

Fenyk, S., Townsend, P. D., Dixon, C. H., Spies, G. B., de San Eustaquio Campillo, A., Slootweg, E. J., …Cann, M. J. (2015). The Potato Nucleotide-Binding Leucine-Rich Repeat (NLR) Immune Receptor Rx1 is a Pathogen Dependent DNA-Deforming Protein. Journal of Biological Chemistry, 290(41), 24945-24960. https://doi.org/10.1074/jbc.m115.672121

• Journal Article Type: Article
• Acceptance Date: Aug 25, 2015
• Online Publication Date: Aug 25, 2015
• Publication Date: Oct 9, 2015
• Deposit Date: Aug 12, 2015
• Publicly Available Date: Oct 23, 2015
• Journal: Journal of Biological Chemistry
• Print ISSN: 0021-9258
• Electronic ISSN: 1083-351X
• Publisher: American Society for Biochemistry and Molecular Biology
• Peer Reviewed: Peer Reviewed
• Volume: 290
• Issue: 41
• Pages: 24945-24960
• DOI: https://doi.org/10.1074/jbc.m115.672121
• Keywords: Cellular immune response, DNA binding protein, Host-pathogen interaction, Nod-like receptor (NLR), Plant biochemistry, Plant defense, Plant virus.
• Public URL: https://durham-repository.worktribe.com/output/1401525