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Title: Structural and biochemical studies of an NB-ARC domain from a plant NLR immune receptor

Abstract

Plant NLRs are modular immune receptors that trigger rapid cell death in response to attempted infection by pathogens. A highly conserved nucleotide-binding domain shared with APAF-1, various R-proteins and CED-4 (NB-ARC domain) is proposed to act as a molecular switch, cycling between ADP (repressed) and ATP (active) bound forms. Studies of plant NLR NB-ARC domains have revealed functional similarities to mammalian homologues, and provided insight into potential mechanisms of regulation. However, further advances have been limited by difficulties in obtaining sufficient yields of protein suitable for structural and biochemical techniques. From protein expression screens in Escherichia coli and Sf9 insect cells, we defined suitable conditions to produce the NB-ARC domain from the tomato NLR NRC1. Biophysical analyses of this domain showed it is a folded, soluble protein. Structural studies revealed the NRC1 NB-ARC domain had co-purified with ADP, and confirmed predicted structural similarities between plant NLR NB-ARC domains and their mammalian homologues.

Authors:
ORCiD logo [1]; ORCiD logo [2]; ORCiD logo [2];  [2]
  1. John Innes Centre, Norwich Research Park, Norwich, England (United Kingdom). Dept. of Biological Chemistry; Lawrence Berkeley National Lab. (LBNL), Berkeley, CA (United States)
  2. John Innes Centre, Norwich Research Park, Norwich, England (United Kingdom). Dept. of Biological Chemistry
Publication Date:
Research Org.:
Lawrence Berkeley National Lab. (LBNL), Berkeley, CA (United States)
Sponsoring Org.:
USDOE Office of Science (SC); Biotechnology and Biological Sciences Research Council (BBSRC) United Kingdom
OSTI Identifier:
1571954
Grant/Contract Number:  
AC02-05CH11231; BB/P012574; BB/J004553
Resource Type:
Accepted Manuscript
Journal Name:
PLoS ONE
Additional Journal Information:
Journal Volume: 14; Journal Issue: 8; Journal ID: ISSN 1932-6203
Publisher:
Public Library of Science
Country of Publication:
United States
Language:
English
Subject:
59 BASIC BIOLOGICAL SCIENCES

Citation Formats

Steele, John F. C., Hughes, Richard K., Banfield, Mark J., and Zeth, Kornelius. Structural and biochemical studies of an NB-ARC domain from a plant NLR immune receptor. United States: N. p., 2019. Web. doi:10.1371/journal.pone.0221226.
Steele, John F. C., Hughes, Richard K., Banfield, Mark J., & Zeth, Kornelius. Structural and biochemical studies of an NB-ARC domain from a plant NLR immune receptor. United States. doi:10.1371/journal.pone.0221226.
Steele, John F. C., Hughes, Richard K., Banfield, Mark J., and Zeth, Kornelius. Wed . "Structural and biochemical studies of an NB-ARC domain from a plant NLR immune receptor". United States. doi:10.1371/journal.pone.0221226. https://www.osti.gov/servlets/purl/1571954.
@article{osti_1571954,
title = {Structural and biochemical studies of an NB-ARC domain from a plant NLR immune receptor},
author = {Steele, John F. C. and Hughes, Richard K. and Banfield, Mark J. and Zeth, Kornelius},
abstractNote = {Plant NLRs are modular immune receptors that trigger rapid cell death in response to attempted infection by pathogens. A highly conserved nucleotide-binding domain shared with APAF-1, various R-proteins and CED-4 (NB-ARC domain) is proposed to act as a molecular switch, cycling between ADP (repressed) and ATP (active) bound forms. Studies of plant NLR NB-ARC domains have revealed functional similarities to mammalian homologues, and provided insight into potential mechanisms of regulation. However, further advances have been limited by difficulties in obtaining sufficient yields of protein suitable for structural and biochemical techniques. From protein expression screens in Escherichia coli and Sf9 insect cells, we defined suitable conditions to produce the NB-ARC domain from the tomato NLR NRC1. Biophysical analyses of this domain showed it is a folded, soluble protein. Structural studies revealed the NRC1 NB-ARC domain had co-purified with ADP, and confirmed predicted structural similarities between plant NLR NB-ARC domains and their mammalian homologues.},
doi = {10.1371/journal.pone.0221226},
journal = {PLoS ONE},
number = 8,
volume = 14,
place = {United States},
year = {2019},
month = {8}
}

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