Large-scale structure-function analysis of the Arabidopsis RPM1 disease resistance protein.
The Arabidopsis RPM1 gene confers resistance against Pseudomonas syringae expressing either the AvrRpm1 or the AvrB type III effector protein. We present an exhaustive genetic screen for mutants that no longer recognize avrRpm1. Using an inducible avrRpm1 expression system, we identified 110 independent mutations. These mutations represent six complementation groups. None discriminates between avrRpm1 and avrB recognition. We identified 95 rpm1 alleles and present a detailed structure--function analysis of the RPM1 protein. Several rpm1 mutants retain partial function, and we deduce that their residual activity is dependent on the level of avrRpm1 signal. In these mutants, the hypersensitive response remains activated if the signal goes above a certain threshold. Missense mutations in rpm1 are highly enriched in the nucleotide binding domain, suggesting that this region plays a key role either in the hypersensitive response associated with RPM1 activation or in RPM1 stability. Cluster analysis of rpm1 alleles defines functionally important residues that are highly conserved between nucleotide binding site leucine-rich repeat R proteins and those that are unique to RPM1. Regions of RPM1 to which no loss-of-function alleles map may represent domains in which variation is tolerated and may contribute to the evolution of new R gene specificities.
Tornero
Pablo
P
Department of Biology, University of North Carolina, Chapel Hill, North Carolina 27599-3280, USA.
Chao
Ryon A
RA
Luthin
William N
WN
Goff
Stephen A
SA
Dangl
Jeffery L
JL
D000483:Alleles; D017360:Arabidopsis / Q000235:genetics* / Q000382:microbiology; D029681:Arabidopsis Proteins; D016000:Cluster Analysis; D004958:Estradiol / Q000494:pharmacology; D005020:Ethyl Methanesulfonate / Q000494:pharmacology; D015870:Gene Expression / Q000276:immunology; D005786:Gene Expression Regulation / Q000187:drug effects; D005816:Genetic Complementation Test; D007113:Immunity, Innate; D016296:Mutagenesis; D020125:Mutation, Missense; D010641:Phenotype; D010935:Plant Diseases / Q000382:microbiology*; D010940:Plant Proteins / Q000235:genetics* / Q000378:metabolism; D011549:Pseudomonas / Q000472:pathogenicity; D014774:Virulence
D029681:Arabidopsis Proteins; D010940:Plant Proteins; C095014:RPM1 protein, Arabidopsis; D004958:Estradiol; D005020:Ethyl Methanesulfonate