plant intracellular innate receptor rpm1 is activated at, and function on, the plasma membrane
TRANSCRIPT
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Zhiyong Gao, Eui-Hwan Chung, Timothy K. Eitas and Jeffery L. Dangl.
PNAS April 13, 2011
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INTRODUCTION
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Introduction
Plant defense against pathogens
MAMPS receptor
Transmembrane pattern
recognition receptors
Plant express
MAMPs
receptors to
detect
microbe
associated
molecularpatterns
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Introduction
Plant defense against pathogens
MAMPS receptors
MAMPS receptor
Recognition
lead to
intracellular
signaling and
transcriptional
output
responses
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Introduction
Nucleotide binding leucine rich repeat (NB-LRR) receptors
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Introduction
Coiled coil nucleotide binding leucine rich repeat (CC NB-LRR)
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Introduction
Nucleotide binding leucine rich repeat (NB-LRR) receptorsHypersensitive-response (HR)
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Introduction
Subcellular localization
Preactivated
NB-LRR
proteins have
been localized
to multiple
subcellular
compartments.
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Introduction
Nucleotide binding leucine rich repeat (NB-LRR) receptors
RPM1 is a CC-NB-LRR protein
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Introduction
Subcellular localization
RPM1
recognize
effectormediated
modifications
of RIN4
AvrRpm1or
AvrB
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MATERIAL AND
METHODS
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Generation ofautoactive alleles
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MATERIAL AND METHODS
Generation of autoactive alleles
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MATERIAL AND METHODS
Generation of autoactive alleles
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MATERIAL AND METHODS
Generation of autoactive alleles
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Vector constructs
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MATERIAL AND METHODS
Vector constructs
pGPTV-HPT
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MATERIAL AND METHODS
Vector constructs
Myc
PCRMyc-tag
N-EQKLISEEDL-C
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MATERIAL AND METHODS
Vector constructs
Myc NES
PCR
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MATERIAL AND METHODS
Vector constructs
MYB nes
PCR
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MATERIAL AND METHODS
Vector constructs
CBL
PCR
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MATERIAL AND METHODS
Vector constructs
gRPM1-myc
T7-RIN4
RPM1-myc induced by estradiol
gRPM1-myc-NES
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Agroinfiltrationand generation oftransgenic plants
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MATERIAL AND METHODS
Agroinfiltration
Agrobacterium
were infiltrated
into leaves of 5 to6 weeks old
Nicotiana
benthamiana
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Conductivitymeasurements
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MATERIAL AND METHODS
Conductivity measurents
Conductivity
measurements
were performedto estimate the
state of the
plant
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RESULTS
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RESULTS
Autoactive alleles analysis
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RESULTS
Autoactive alleles analysis
Affected in the
coiled coil domain
Affected in spacer
region
Affected in the
P-loop domain
Lost of function mutants
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RESULTS
Autoactive alleles analysis
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RESULTS
Autoactive alleles analysis
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RESULTS
Autoactive alleles analysis
The disappearance of RPM1 is correlated with effector-
mediated activation
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RESULTS
RIN4 role as supressor
RPM1 wild type
is weakly
autoactive in a
rin4 null mutant
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RESULTS
RIN4 role as supressorRIN4 expression delayed and diminished cell death
RPM1(D505V) + rin4
5
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RESULTS
RIN4 role as supressor
The transgenic lines gRPM1(D505V) rmp1
rin4 were difficult to recover
g
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RESULTS
Localization of RPM1
RPM1 is associated to membranes
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MATERIAL AND METHODS
Cellular localization
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RESULTS
Localization of RPM1
RPM1 is predominantly localized on plasmatic membrane
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RESULTS
Localization of RPM1
It is highly unlike that activated RPM1
relocalizes to the nucleus
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RESULTS
Localization of RPM1
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MATERIAL AND METHODS
Generation of autoactive alleles RESULTS
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Localization of RPM1
RPM1 neither relocalizes to, nor
function in the nucleus
MATERIAL AND METHODS
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Generation of autoactive alleles RESULTS
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Localization of RPM1Plasma membrane-tethered RPM1 retains HR function
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CONCLUSIONS
CONCLUSIONS
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4) The results indicate that activated RPM1 resides
solely on the plasma membrane.
3) Preactivated RPM1 resides on the plasma
membrane.
1) The P-loop is important for RPM1 function and
localization.
2) The CC domain is critical for function.