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GIGANTEA mediated salt stress GIGANTEA-mediated salt stress signaling in plant g g p
(식물에서 자이겐티아에 의한 내염성 기작 조절)
Woe Yeon Kim
World Class University Program and Department of Applied life chemistry Department of Applied life chemistry,
Gyeongsang National University
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GIGANTEA (GI)GIInput
- photomorphogenesis
GIGANTEA (GI)
OscillatorLHY TOC1
photomorphogenesis, - photoperiodic flowering regulation- circadian clock regulation
OscillatorLHYCCA1
TOC1g
- excess starch accumulation- alterations to sucrose metabolism
lt d iti it t GI
- altered sensitivity to Light, Cold, and Oxidative stress
CO
Output(photoperiodicfl in C d f 1173
FTflowering pathway)
~ Composed of 1173 a.a.
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GIGANTEA controls the circadian clock and time of flowering through protein stabilizationtime of flowering through protein stabilization
Modified from Deng (2012) Trends in Plant Science
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Salt stress inhibits plant growth and delays p g yflowering time in Arabidopsis plants.
Xua Li et al. (2007) Plant Growth Regul.
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Floral transition in Arabidopsis
GI
TOC1
ZTL GI FKF1
CDF1
CO
LFY
Modified from Blázquez et al. (2001) EMBO reports
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CO plays a key role in salt-induced late floweringp y y f g
Xua Li et al. (2007) Plant Growth Regul.
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Does GI play a role in salt induced late flowering ?
300 mM25 mM
20
g tim
em
ber)
25 mM
10
Flow
erin
g(le
af n
um
WT GI-OXgi-1 WT GI-OXgi-10 WT GI-OXgi-1
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GI is degraded upon salt stress
Transcript 0 12 24 (hr)
Protein 0 12 24 (hr)
3 ControlNaClMG132NaCl+MG132n
(fold
)
( )GITUBGI-HA
( )Control
NaCl2
NaCl+MG132
GI p
rote
in
MG132NaCl+MG132
0
1
Rel
ativ
e G
00 12 24
Time (hr)
R
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GI mutant exhibits increased salt tolerance
WT GI-OX gi-1
0 mM
A
ed
B
4
5150mM NaCl
ht (g
)
0 mM
plan
ts T
reat
ew
o w
eeks
2
3
resh
wei
g150 mM
250 Mw
eeks
old
pith
sal
t for
tw0
1Fr
WT GI-OX gi-1
250 mMTh
ree w
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Salt stress tolerance in Plants
[Ca 2+]ext
High [Na +]
SOS1
H +Sodium efflux
[Ca 2+]int
PM
Ion SOS3 SOS2
SOS1
Na +
Na +
[ ]int
homeostasisSalt stress signaling
H + H +
Ca 2+
V-ATPasePPase
MYC/MYB/AREB/DREB2A
NU
Compartmentation
NHX
VacuoleH +
N + 2
RD22, ADH, RD29a/bP5CS,BADH
Na +
Na +
ABI2
Ca 2+
SOS3SOS2
Osmolytes(Proline, betaine)?
K +
HKT1Na
Receptor
Modified from Cheong and Yun (2007) J. Plant Biol
betaine)
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Does GI regulate general stress pathway ?
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Salt stress tolerance in Plants
[Ca 2+]ext
High [Na +]
SOS1
H +Sodium efflux
[Ca 2+]int
PM
Ion SOS3 SOS2
SOS1
Na +
Na +
[ ]int
homeostasisSalt stress signaling
H + H +
Ca 2+
V-ATPasePPase
MYC/MYB/AREB/DREB2A
NU
Compartmentation
NHX
VacuoleH +
N + 2
RD22, ADH, RD29a/bP5CS,BADH
Na +
Na +
ABI2
Ca 2+
SOS3SOS2
Osmolytes(Proline, betaine)?
K +
HKT1Na
Receptor
Modified from Cheong and Yun (2007) J. Plant Biol
betaine)
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SOS1 i l b N /H hSOS1 is a plasma membrane Na+/H+ exchanger
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GI mutant plasma membrane vesicles exhibits high Na+/H+ antiporter activityNa /H antiporter activity
20gi-1
15
n-1
mg-
1 gi-1Col-0sos1-1
+NaCl
5
10
/Fm
axm
i
0
5∆F/
0 50 100 150 200 250NaCl (mM)
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GI influences SOS1 stability
SOS1WB:
SOS1 Anti-SOS1
CBBCBB
SOS1Transcript
TUB
p
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GI mutants exhibits increased salt tolerance
ore
ter
Bef
o
mM
Aft
150
mM
0
1
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GIGANTEA controls the circadian clock and time of flowering through protein stabilizationtime of flowering through protein stabilization
Modified from Deng (2012) Trends in Plant Science
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Sites for GI to regulate salt stress ?
Hi h [N +]High [Na +]
SOS1
Na +
H +
GI?Ca 2+ SOS3
SOS2GI?GI?
GI?
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GI interaction with SOSs proteins
DIC YFP MERGE
GI-VN +VC-SOS2
GI-VN +VC-SOS2
GI-VN +VC
VN +VC SOS2
C I i it ti
VC-SOS2
Bimolecular FluorescenceCo-Immunoprecipitation assay Bimolecular Fluorescence Complementation (BiFC)
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GI interacts SOS2 directly in vitro
Input S2 S3
35S- GISOS1
H +
GST
35S-
GI
Input
GST
-SO
S
GST
-SO
S
SOS3SOS2
GI?GI?Na +
35S- GI High [Na +]
Ca 2+
CBB
GST- SOS2
GST- SOS3H +
GSTSOS3
SOS1
Na +
High [N +]
Ca 2+SOS2
GI[Na +]
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SOS1 is phosphorylated by SOS2 upon salt stress
SOS1:His
Quintero et al. (2002) PNAS 99: 9061-9066
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GI inhibits SOS2 kinase activity
12)
68
1012
sity
(SO
S1)
246
ativ
e in
tens
0Rel
a
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gi-1 salt tolerance and SOS1 stability is SOS2 dependentdependent
ore
Bef
oM
Afte
r m
M
0 m
M15
0 m
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Sites for GI to regulate salt stress ?
Hi h [N +]High [Na +]
SOS1
Na +
H +
GI?Ca 2+ SOS3
SOS2
GI?
SOS2
GI
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GI/SOS2 interaction is abolished by SOS3 and Salt
Hi h [N +]High [Na +]SOS1
SOS3
GISOS2SOS2
GI SOS3
100 mM NaCl / 24 h-NaCl
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Working Model
High [Na +]
- SALT +SALTSalt tolerance g [ ]
SOS1
Na +
H +
SOS1 P
Ca 2+
SOS3 SOS2SOS3
GISOS3
SOS2
Inhibition
SOS2SOS3
GI GI