fudan sugar signaling 8 15 14 final植物糖代谢及营养信号传导

植物糖代谢及营养信号传导

TOR激酶作为分子开关调控植物生长和抗逆的新机制

熊

延

植物逆境生物学

ntsaresoimportant!

Food

Industry Energy

Cleanairandwater

Livestockfeed

Medicines

Shelter

Textiles

2

EnergySignalinginPhotosynthesisnts

SolarEnergyCO2

Water

O2

Sugars,COrganicN

h

3

biibl

Unbelievableasitseems,accordingtotheUSDA,theaverageAmericanconsumesthefollowingeachyear:

134poundsofrefinedsugarexcludinghoney

365servingsofsodapop(638cansperyearforpeopleaged12-29)

200sticksofgum

22poundsofcandy

63dozendoughnuts

60poundsofcakesand s

23gallonsoficecream

ntsneedtomodulateandcoordinate

sugarsignalsprecisely

5

Roleofsugarsin ntgrowthanddevelopment

GlucosePromotion&Repression

ofntGrowth

Glc00.40.8120/Hlight23456%

Sugarcontrols:

Juveniletoadultphasetransition

Senescence

Stressadaption..

8

Molecularmechanismsofsugarregulation

Transcriptioncontrol,egphotosynthesisgenes

Transcriptsstabilityandprocessing

Translation

Proteinstability(EIN3-P;Autophagy…..)

HowSugarsperformthesefunctions?

植物糖营养信号传导

Cross-talkbetweensugarandhormones

GlucoseSensor(HXK1)

Centergrowthregulator(TOR)

植物糖营养信号传导

Cross-talkbetweensugarandhormones

GlucoseSensor(HXK1)

Centergrowthregulator(TOR)

2

Todissectsugarsignalingnetwork

3

ArabidopsisGlucoseSignalingMutants

gin:glucoseinsensitiveglo:glucoseoversensitive

5

CrosstalkbetweenSugar,ABAandET

Leonetal.,2003

CrosstalkbetweenSugar,ABAandET

Leonetal.,2003

Asastresshormone,ABAcanalsopromotegrowth

CrosstalkbetweenSugar,AuxinandCytokinin

Mooreetal.,2003Science

GlucoseasCentralRegulatorySignal

Glucose&HormoneConnections

Sugarsfunctionsas

Signalingmolecules

Metabolcpathways

2

SugarSensingSystems

F-6-P

TOR

Rollandetal.,2006AnnualRevntBio

Fuetal.2014Cell

22

植物糖营养信号传导

Cross-talkbetweensugarandhormones

GlucoseSensor(HXK1)

Centergrowthregulator(TOR)

UncoupleSignalingfromMetabolism

gin2,GlucoseSensor(HXK1)

24

GlucoseRepression

gin2IsaHXK1Mutant

WTgin2

WTOVgin2HXK1/gin2

25

GlucosePromotion

HXK1modulates

ntgrowth&developmentinresponsetoenvironment

26

AtHXK1isaninternalfactorforArabidopsisgrowth

gin2IsaHXK1Mutant

Thefirststepinthereactionsofglycolysis

Hexokinase

Glucose Glucose-6-Phosphate

ATPADP

HowaboutHXK1catalyticallyinactivemutants?

27

AtHXK1isaninternalfactorforArabidopsisgrowth

gin2IsaHXK1Mutant

Thefirststepinthereactionsofglycolysis

Hexokinase

Glucose

Glucose-6-Phosphate

ATPADP

CatalyticallyinactivemutantsG104D:ATPbinding

S177A:phosphortransfer

Mooreetal.,Science20023

GlucoseSignalinginLeafCells

Mooreetal.,Science20023

HXK1ActivityinTransgenicnts

Mooreetal.,Science20033

UncoupleGlucoseSignalingfromMetabolism

GlucoseRepressionofGrowth

Glc

Man

WTgin2HXK1S177AG104D

Mooreetal.,Science20033

UncoupleGlucoseSignalingfromMetabolism

GlucosePromotionofGrowth

WT

-

gin2

- HXK1S177AG104D

Mooreetal.,Science20033

HowHXK1functionsasaglucosesensor?

NuclearGlucoseSensor

3

NuclearGlucoseSensing&Signaling

34

NuclearHXK1

HXK1p::HXK1-GUS/gin2 HXK1p::HXK1-GFP/gin2

35

IsolationofHXK1nuclearcomplex

Choetal.,2006Cell3

HXK1,HUP1&HUP2

NuclearProteinComplex

HUP2

THUP

T

HXK1

IP(HXK)IP(HUP1)

TNSNSSS

anti-HXK

anti-HUP1

HUP1/VHA-B1

anti-VHA-A

anti-HUP2

HUP2/RPT5B

anti-RPT1

HXK/gin2WT WT

Transgenic

HUP:HXK1UnconventionalPartner

Choetal.,2006Cell

3

7

P(HUP1)

IP(HUP2)

HUP:HXK1UnconventionalPartner

HUP1,VHA-B1:BsubunitofV1ComplexinV-ATPasevacuoles,ER,Golgiandsmamembrane

HUP2,RPT5B:onecomponentofproteasomeCytosol

Choetal.,2006Cell

38

HUP1&HUP2MediateBothGlucoseRepression&Promotion

GR

MC

GP

gin2Colhup1hup2

Colhup1hup2Lergin2

2%Glc

Choetal.,2006Cell39

NuclearHXK1-HUPComplexonSpecificTargetGenePromoters

2%Glc

2%Glc

ChIP

Choetal.,2006Cel4l0

TheHXK1NuclearProteinComplexes

TFs

HUP1

HXK1

HUP2

TFs

4

GlucoseSensors

Arabidopsis

Yeast

Humans

DiabetesObesityCancerLongevity

42

植物糖营养信号传导

Cross-talkbetweensugarandhormones

GlucoseSensor(HXK1)

Centergrowthregulator(TOR)

43

44

Autotrophictransitionaftergermination

Photosynthesis

heterotrophic

autotrophic

Glucosecontrolsrootmeristemactivation

Photosthesis

LeafSource

Zhangetal.2010

RootSink

-Glc

-Glc

-Glc

47

50

3-day-germinationundernoglucoselowlight1dayglucose(15mM)treatment

WThxk1

Glc- -

WThxk1

-

Glc-

HXK1dependent?

RootmeristemactivationisnotcontrolledbyHXK1

Rootmeristemsize

EdU

EdU

Rootmeristemduringautotrophictransition

LeafSource

DZ

EZ

Seed

StemcellnicheQuiescentcenter

RootSink

MZ

MZ:meristemzoneEZ:elongationzone

DZ:differentiationzone

46

Howdoesglucoseregulaterootmeristemactivation?

48

Glucosehastwoprimaryrolesingrowthanddevelopment

MetabolicSupply

Glucose

SignalMolecule

Glose

Glycolysis

Hexikinase1(HXK1)

Pyruate

TCAcycleNADHFADH2

ElectronTransportChain

OxidatiePhsphorylation

NucleusGeneexpression

ATP

Mitochondria

MooeetalScience2003Choetal.Cel2006

49

Glucose-glycolysis-mitochondriaenergyrelay

controlsrootmeristemactivation

Metabolicsupply?

Glc

--2-DGAADNP

2-DG

Glc

--2-DGAADNP

AA

DNP

2-DG2-deoxyglucoseglycolysisblocker

AAantimycnAmitochondrialelectrontransportinhibitor

DNP24-dnitrophenolmitochondrialuncoupler

5

Howisthismetabolicsignaltransduced?

?

52

Howisthismetabolicsignaltransduced?

M ls

GrowthfactorsStressLowEnergyGlucose

AA

AKTMAPK

AMPK

TSC1/2

RAGs

Rapt

mTOR

LST8

Rapamycin

FKBP12

4E-BPS6K

Ribosomebiogeness

TranslationAutophagy

TOR

TOR:TargetofRapamycin

53

Howisthismetabolicsignaltransduced?

WT

- Raptor

Glc-

TOR

54

Glucose-TORsignalingcontrolscellproliferation

Col tor

GlcGlcRapGlcAAGlc

After24hoursglucosetreatment

Rap:rapamycinAA:antimycinA

55

Glc-TORdirectsspecifictranscriptionalnetworks

3daysundernoglucoselowlight

2hoursglucose(15mM)treatment

WTtor

Glucose

GycoyssTCAcyceETC

ATP

TORkinase

Metabolictranscriptionnetwork

Cellcycle&DNAsynthesis

Gycoyss Proendegradaton

TCAcyce&ETC Lpddegradaton

Rbosomebogeness Amnoacddegradaton

Protensynthess Autophagy

Amnoacdynthess CHOmetabosm

Nuceotdesynthess Bodegradatonofxenobotcs

RNAsynthess&processng GuconeogeneseTFs&Chromatnmoduators Deveopment

Sgnang TFs&Chromatnmoduators

Transporters Stress

Cewasynthess Transporters

Lgnnynthess

GucosnoatesynthessS-assmaton

P<0.01;Log2≥1

6

S-phasegeneactivationrequiresTOR-directlyphosphorylationofE2Fa

E2Fa-Flag-

α-Flag-

IPα-FlagWα-TOR

Wα-TOR

Wα-Flag

Torin1- -101001000(nM)TOR-

32P-ATP

E2Fa

E2FA1

Δ11Δ2

Δ3

Δ480

485

162

162

420

420

485

420

WT123

3

**** ***

2P-ATP

**************

***

E2Fa-6HIS

7

WTΔ1Δ2Δ3Δ

Glucose-TORsignalingdependsE2Fa

WTe2fa

WT2f

Glc- -

3DAG1DGlc

Glc/-Glc

* *

*

**

**

*

58

E2Fa

PstDC3000

1318genes 1050genes

Foldchanges

EdU

EdU

Foldchanges

Glucose-TORsignalinginrootmeristemactivation

LeafSourcephotosynthesissucrose/glucose

glucose

glycolysismitochondriaETC>OXP>ATP

TORsignaling

Meristem Sphase

RootSink

XongandSheen,JBC,2012Xongetal.,Nature,2013

XongandSheen,CellCycle,2013

XongandSheen,ntPhysology,2014

59

AtTORdown-regulatesdefense

22:aconserved

ofbacterialflagellin

MAMPs:Microbe-AssociatedMolecularPatterns

60

6

Summary

TOR

MKKs(MKK4,5)

Innateimmunity

Cellproliferation

MPK3/6

E2Fa

MTKs(ANP1/2)

Favorablegrowthconditions

62

Summary

TOR

MKKs(MKK4,5,9)MPK3/6

Innateimmunity

Cellproliferation

E2Fa

MTKs(ANP1/2)

Unfavorablegrowthconditions

Growthanddefensearetightlyconnected

Favorableconditions

Lessresistant

abioticstresses

Moreresistant

bioticstresses

UncouplegrowthanddefensebymodifyingTOR

downstreameffectors?e.g.E2Fs,ANP1/2?

63

ntsSupplyCarbon,NitrogenandOxygen

drought

flood

fungi

pollution

bacteria

oomycetes

Growth

development

temperature

Bioticstresses

Abioticstresses

TOR

kinase

PhotosynthesisNitrateassimilation

Adoorhasjustbeenopened…

Takehomemessages

Sugarsignalingisverycomplex

Crosstalkedwithhormones

Sensedbydifferentmechanisms

Newsignalingfunctionsofmetabolicenzymes

TORasacentralregulatorforgrowthanddefense

参考文献

FuY.,LimS.,UranoD.,Tunc-OzdemirM.,PhanN.,ElstonT.,Jones,AM.2014Reciprocalencodingofsignalintensityanddurationinaglucose-sensingcircuit.Cell156:1084-1095

LastdragerJ.,HansonJ.,SmeekensS.2014Sugarsignalsandthecontrolofntgrowthanddevelopment.J.Exp.Bot10.1093/kxb/ert474

Xiong,Y.,McCormack,M.,Li,L.,Hall,Q.,Xiang,C.andSheen,J.2013Glucose-TOR

signalingreprogramsthetrascriptomeandactivatesmeristems:10.1038/nature12030

YuS.,CaoL.,ZhouCM.,ZhangT.,LianH.,SunY.,WuJ.,HuangJ.,WangG.,WangJW.

2013Sugarisanendogenouscueforjuvenile-to-adultphasetransitioninntsElfe

LiP.,WindJ.,ShiX.,ZhangH.,HansonJ.,SmeekensSC.,TengS.2011.FructosesensitivityissuppressedinArabidopsi