First published online October 12, 2007; 10.1105/tpc.106.048801
The Plant Cell 19:3297-3313 (2007)
© 2007 American Society of Plant Biologists
Identification and Mutational Analysis of Arabidopsis FLS2 Leucine-Rich Repeat Domain Residues That Contribute to Flagellin Perception[W]
F. Mark Dunninga,b,
Wenxian Suna,
Kristin L. Jansena,
Laura Helfta,b and
Andrew F. Benta,b,1
a Department of Plant Pathology, University of Wisconsin, Madison, Wisconsin 53706
b Cell and Molecular Biology Program, University of Wisconsin, Madison, Wisconsin 53706
1 Address correspondence to afb{at}plantpath.wisc.edu.
Mutational, phylogenetic, and structural modeling approaches were combined to develop a general method to study leucine-rich repeat (LRR) domains and were used to identify residues within the Arabidopsis thaliana FLAGELLIN-SENSING2 (FLS2) LRR that contribute to flagellin perception. FLS2 is a transmembrane receptor kinase that binds bacterial flagellin or a flagellin-based flg22 peptide through a presumed physical interaction within the FLS2 extracellular domain. Double-Ala scanning mutagenesis of solvent-exposed ß-strand/ß-turn residues across the FLS2 LRR domain identified LRRs 9 to 15 as contributors to flagellin responsiveness. FLS2 LRR-encoding domains from 15 Arabidopsis ecotypes and 20 diverse Brassicaceae accessions were isolated and sequenced. FLS2 is highly conserved across most Arabidopsis ecotypes, whereas more diversified functional FLS2 homologs were found in many but not all Brassicaceae accessions. flg22 responsiveness was correlated with conserved LRR regions using Conserved Functional Group software to analyze structural models of the LRR for diverse FLS2 proteins. This identified conserved spatial clusters of residues across the ß-strand/ß-turn residues of LRRs 12 to 14, the same area identified by the Ala scan, as well as other conserved sites. Site-directed randomizing mutagenesis of solvent-exposed ß-strand/ß-turn residues across LRRs 9 to 15 identified mutations that disrupt flg22 binding and showed that flagellin perception is dependent on a limited number of tightly constrained residues of LRRs 9 to 15 that make quantitative contributions to the overall phenotypic response.
This article has been cited by other articles:
|
|
|
|
 
H. Haweker, S. Rips, H. Koiwa, S. Salomon, Y. Saijo, D. Chinchilla, S. Robatzek, and A. von Schaewen
Pattern Recognition Receptors Require N-Glycosylation to Mediate Plant Immunity
J. Biol. Chem.,
February 12, 2010;
285(7):
4629 - 4636.
[Abstract]
[Full Text]
[PDF]
|
|
|
|
|
|
|
G. Wang, Y. Long, B. P.H.J. Thomma, P. J.G.M. de Wit, G. C. Angenent, and M. Fiers
Functional Analyses of the CLAVATA2-Like Proteins and Their Domains That Contribute to CLAVATA2 Specificity
Plant Physiology,
January 1, 2010;
152(1):
320 - 331.
[Abstract]
[Full Text]
[PDF]
|
|
|
|
|
|
|
L. Feiz, B. S. Beecher, J. M. Martin, and M. J. Giroux
In Planta Mutagenesis Determines the Functional Regions of the Wheat Puroindoline Proteins
Genetics,
November 1, 2009;
183(3):
853 - 860.
[Abstract]
[Full Text]
[PDF]
|
|
|
|
|
|
|
V. Nicaise, M. Roux, and C. Zipfel
Recent Advances in PAMP-Triggered Immunity against Bacteria: Pattern Recognition Receptors Watch over and Raise the Alarm
Plant Physiology,
August 1, 2009;
150(4):
1638 - 1647.
[Full Text]
[PDF]
|
|
|
|
|
|
|
M. Casasoli, L. Federici, F. Spinelli, A. Di Matteo, N. Vella, F. Scaloni, J. Fernandez-Recio, F. Cervone, and G. De Lorenzo
Integration of evolutionary and desolvation energy analysis identifies functional sites in a plant immunity protein
PNAS,
May 5, 2009;
106(18):
7666 - 7671.
[Abstract]
[Full Text]
[PDF]
|
|
|
|
|
