植物生理与分子生物学：植物糖代谢及营养信号传导

植物糖代谢及营养信号传导FoodCleanairandwaterShelterEnergyLivestockfeedIndustryMedicinesTextilesPlantsaresoimportant!/EnergySignalinginPhotosynthesisPlantsSolarEnergyCO2

WaterO2Sugars,COrganicNUnbelievableasitseems,accordingtotheUSDA,theaverageAmericanconsumesthefollowingeachyear:134poundsofrefinedsugarexcludinghoney365servingsofsodapop(638cansperyearforpeopleaged12-29)200sticksofgum22poundsofcandy63dozendoughnuts60poundsofcakesandcookies23gallonsoficecreamPlantsneedtomodulateandcoordinatesugarsignalspreciselySugar

Metabolism

and

Signals

in

Source

CellsSugar

Metabolism

and

Signals

in

Sink

cellsRole

of

sugars

in

plant

growth

and

developmentGlucosePromotion&RepressionGlc00.40.8120/Hlight23456%ofPlantGrowthSugar

controls

juvenile

to

adult

phase

transitionYuetal.,2013ElifeSugarcontrols:SenescenceStressadaption…..MolecularmechanismsofsugarregulationTranscriptioncontrol,e.g.photosynthesisgenesTranslationTranscriptsstabilityandprocessingProteinstability(EIN3-P;Autophagy…..)HowSugarsperformthesefunctions?Cross-talkbetweensugarandhormonesGlucoseSensor(HXK1)FructoseSensor(FINS1)EnergySensor(KIN10/11)Centergrowthregulator(TOR)OR)

植物糖营养信号传导Cross-talkbetweensugarandhormonesGlucoseSensor(HXK1)FructoseSensor(FINS1)EnergySensor(KIN10/11)Centergrowthregulator(TOR)OR)

植物糖营养信号传导To

dissect

sugar

signaling

networkArabidopsisGlucoseSignalingMutantsgin:glucoseinsensitiveglo:glucoseoversensitive

CrosstalkbetweenSugar,ABAandETLeonetal.,2003Leonetal.,2003CrosstalkbetweenSugar,ABAandETAsstresshormones,ABAcanalsopromotegrowthCrosstalkbetweenSugar,AuxinandCytokininMooreetal.,2003ScienceGlucoseasCentralRegulatorySignalGlucose&HormoneConnectionsSugarsfunctionsasMetabolicpathwaysSignalingmoleculesSugarSensingSystemsRollandetal.,2006AnnualRevPlantBioFuetal.,2014CellF-6-PTORCross-talkbetweensugarandhormonesGlucoseSensor(HXK1)FructoseSensor(FINS1)EnergySensor(KIN10/11)Centergrowthregulator(TOR)OR)

植物糖营养信号传导

UncoupleSignalingfromMetabolism

gin2,GlucoseSensor(HXK1)

GlucoseRepression

gin2IsaHXK1MutantWTOVgin2HXK1/gin2WTgin2

GlucosePromotionHXK1modulatesplantgrowth&developmentinresponsetoenvironment

AtHXK1isaninternalfactor

forArabidopsisgrowthThefirststepinthereactionsofglycolysisGlucose-6-PhosphateGlucoseHexokinaseATPADPHowaboutHXK1catalyticallyinactivemutants?gin2IsaHXK1MutantAtHXK1isaninternalfactor

forArabidopsisgrowthThefirststepinthereactionsofglycolysisGlucose-6-PhosphateGlucoseHexokinaseATPADPCatalyticallyinactivemutantsG104D:ATPbindingS177A:phosphortransferMooreetal.,Science2003gin2IsaHXK1MutantGlucoseSignalinginLeafCellsMooreetal.,Science2003HXK1ActivityinTransgenicPlantsMooreetal.,Science2003UncoupleGlucoseSignalingfromMetabolism

GlucoseRepressionofGrowthWTgin2HXK1S177AG104DGlcManMooreetal.,Science2003

WT

-

-HXK1S177AG104D

gin2UncoupleGlucoseSignalingfromMetabolism

GlucosePromotionofGrowthMooreetal.,Science2003WTgin2HXK1S177AG104DIAACotHypRootUncoupleGlucoseSignalingfromMetabolism

CellExpansion&ProliferationMooreetal.,Science2003

HowHXK1functionsasaglucosesensor?NuclearGlucoseSensor

NuclearGlucoseSensing&SignalingHXK1p::HXK1-GUS/gin2HXK1p::HXK1-GFP/gin2NuclearHXK1IsolationofHXK1nuclearcomplexChoetal.,2006Cellanti-HXKanti-HUP1anti-VHA-Aanti-HUP2anti-RPT1HXK/gin2TransgenicIP(HXK)TNSIP(HUP1)NSWTWTIP(HUP1)IP(HUP2)SSHXK1,HUP1&HUP2NuclearProteinComplexHXK1HUP1HUP2TF1TF2HUP1/VHA-B1HUP2/RPT5BHUP:HXK1UnconventionalPartner

Choetal.,2006CellHUP1,VHA-B1:BsubunitofV1ComplexinV-ATPase

vacuoles,ER,GolgiandplasmamembraneHUP2,RPT5B:onecomponentofproteasomeCytosolHUP:HXK1UnconventionalPartner

Choetal.,2006CellColhup1hub2gin2Colhup1hup2

HUP1&HUP2MediateBothGlucoseRepression&Promotion

GRGPMCColhup1hup2Lergin22%GlcChoetal.,2006Cell

NuclearHXK1-HUPComplexonSpecificTargetGenePromoters

2%GlcChIP2%GlcChoetal.,2006CellTheHXK1NuclearProteinComplexesHXK1HUP1HUP2TFsTFsGlucoseSensorsArabidopsisYeastHumansDiabetesObesityCancerLongevityCross-talkbetweensugarandhormonesGlucoseSensor(HXK1)FructoseSensor(FINS1)EnergySensor(KIN10/11)Centergrowthregulator(TOR)OR)

植物糖营养信号传导FructosesignalinginplantGlucose-likeinhibitioneffect,butHXK1independentChoetal.,2011CrosstalkbetweenfructosesignalingandABA,ETChoetal.,2011ScreeningofputativefructosesensorChoetal.,2011FINS1/FBPinfructosesignalingChoetal.,2011FINS1/FBPlocalizesbothincytoplasmandnucleusSimilarregulatoryroleongeneexpressionasHXK1?Choetal.,2011FructosesensingQTLLietal.,2011PNASFSQ6isspecificallysensitivetofructoseLietal.,2011PNASFSQ6/ANAC089isanucleartranscriptionfactorLietal.,2011PNASCross-talkbetweensugarandhormonesGlucoseSensor(HXK1)FructoseSensor(FINS1)EnergySensor(KIN10/11)Centergrowthregulator(TOR)OR)

植物糖营养信号传导STRESS>50%REDUCTIONINCROPYIELDshareddownstreamresponsesSIMILARDOWNSTREAMRESPONSES*RELEVANCEOFTHISCOMPONENT(S)TOOTHERASPECTSOFPLANTLIFE?*COMMONSIGNAL?*SENSOR(S)–TRANSDUCER(S)OFSIGNAL?DIFFERENTSTRESSES

CO2H20RESPIRATIONPHOTOSYNTHESISGLUCOSEO2

SUNLIGHTATPATPFLOODINGDARKDROUGHTPOLLUTANTSINDUCEDBYDARKNESSDINDINDIN

GENES+SUGARDINREPRESSEDBYSUGARMETABOLICVS.LIGHTSIGNAL?

LinkingDarkness,Herbicide,Hypoxia,Stress…

NutrientDeficiency

DIN1,DIN6GlucosecanrepresstheDIN1,DIN6inductionHXK1independentBaena-Gonzálezetal.,Nature,2008SIMILARDOWNSTREAMRESPONSES*RELEVANCEOFTHISCOMPONENT(S)TOOTHERASPECTSOFPLANTLIFE?*COMMONSIGNAL?*SENSOR(S)–TRANSDUCER(S)OFSIGNAL?DIFFERENTSTRESSES

AMPKPLANTHOMOLOGUEOFSNF1(YEAST)ANDAMPK(MAMMALS)ISAGOODCANDIDATESTARVATIONAMPAMPKGlucosetransportCatabolicpathwaysCellsurvivalAMPK,AMP-ACTIVATEDKINASE,MAMMALSBiosyntheticpathwaysTranslationREGULATIONOFENZYMEACTIVITYCANCERDIABETESOBESITYFunctionalScreenofSnRKs

EnergySensors/CIPK/38PKsGenomiccDNA(RNA)cDNA(RNA)DNA seedlingsprotoplastsAKIN10AKIN11AKIN12AKIN10AKIN11AKIN12AKIN10AKIN11AKIN12EXPRESSIONANALYSESOFSNRK1GENESDOSnRKSREGULATEJUSTASETOFBIOSYNTHETICENZYMES?DOTHEYHAVEABROADERROLEEXERTEDTHROUGHGENEEXPRESSION?ANewRoleofKIN10/KIN11inTranscriptionalControl

Baena-Gonzálezetal.,Nature,2008INDUCEDGENESREPRESSEDGENESKIN10KIN10-INDUCEDCHANGESSTRESS,DARKNESS,STARVATION“LOWSUGAR”SIMILAR“HIGHSUGAR”MAXIMALPHOTOSYNTHESIS,SUGARFEEDINGOPPOSITEKIN10IsaMasterRegulatorEnergyhomeostasis,Growth,Stressresponse,SurvivalDevelopment,Reproduction,Senescence

NUTRIENTSTARVATIONUpstreamPKsS-groupbZIPsTFsChromosome/histonemodificationSignalling(hormones,PKs,PPs,Ca2+,photoreceptors)Protein,aminoacid&lipiddegradationCellwall,starch&sucrosedegradationAutophagyPhotosynthesisGluconeogenesisTrehalosemetabolismTransportersProteinsynthesisTFsChromosome/histonemodificationSignalling(hormones,PKs,PPs,Ca2+)AminoacidbiosynthesisCellwallsynthesisNucleotidemetabolismTCAcycleGlycolysisHMG-CoAR,SPS,NR,TPS5,AGPasePost-translationalControlGlucoseSucrosePPsAMPSTRESSEnergydeficitSnRK1Stress-specificresponsesPTFsTFsTRANSCRIPTIONALREPROGRAMMINGBiosynthesisUPDOWNDARKNESSConvergentStress&EnergySignalingBaena-Gonzálezetal.,Nature,2008KIN10PROMOTESSTRESSTOLERANCEO1O2CINCREASEDTOLERANCETOSTARVATIONSUGARDEPLETIONFROMMEDIAO1O2CPIGMENT0123LESSSTRESS-RELATEDPIGMENTATIONBaena-Gonzálezetal.,Nature,2008O1O2CDELAYEDGROWTHANDAGEINGKIN10DELAYSGROWTHANDAGEINGO1CPROLONGUEDJUVENILEPHASEBaena-Gonzálezetal.,Nature,2008d-1d-2C10-110-211O1O2d-1C10-111O1d-2O2KIN10-KIN11SILENCINGCAUSESGROWTHARRESTANDPREMATUREAGEING4WEEKS7WEEKSBaena-Gonzálezetal.,Nature,2008

TargetGenesofGlucose&EnergySensorsHXK1HXK1KIN10HXK1KIN10HowKIN10performitsenergysensorfunction?TheKIN10NuclearProteinComplexesACTFsTFsKIN10

a

gbRC?NCross-talkbetweensugarandhormonesGlucoseSensor(HXK1)FructoseSensor(FINS1)EnergySensor(KIN10/11)Centergrowthregulator(TOR)OR)

植物糖营养信号传导AutotrophictransitionaftergerminationheterotrophicautotrophicPhotosynthesisRootmeristemduringautotrophictransitionLeafSourceRootSinkSeedMZEZDZQuiescentcenterStemcellnicheMZ:meristemzoneEZ:elongationzoneDZ:differentiationzoneGlucosecontrolsrootmeristemactivationZhangetal.,2010-Glc+GlcLeafSourceRootSinkPhotosynthesis-Glc+GlcRootmeristemsizeEdU-Glc+GlcHowdoesglucoseregulaterootmeristemactivation?GlucosehastwoprimaryrolesingrowthanddevelopmentSignalMoleculeGlucoseHexikinase1(HXK1)NucleusGeneexpressionMetabolicSupplyGlucoseGlycolysisPyruvateTCAcycleNADH,FADH2ElectronTransportChainOxidativePhosphorylationATPMitochondriaMooreetal,Science2003Choetal.,Cell2006RootmeristemactivationisnotcontrolledbyHXK1HXK1dependent?-+-+GlcWThxk13-day-germinationundernoglucose,lowlight1dayglucose(15mM)treatment-+-+GlcWThxk1EdUGlucose-glycolysis-mitochondriaenergyrelaycontrolsrootmeristemactivationMetabolicsupply?2-DGAA2-DG:2-deoxyglucose,glycolysisblockerAA:antimycinA,mitochondrialelectrontransportinhibitorDNP:2,4-dinitrophenol,mitochondrialuncouplerDNP--2-DGAADNP+Glc--2-DGAADNP+GlcHowisthismetabolicsignaltransduced??Howisthismetabolicsignaltransduced?TORRaptorLST8mTORFKBP12RapamycinMAPKAMPKGrowthfactorsStressLowEnergyGlucoseAKTS6KTranslationAutophagyRibosomebiogenesis4E-BPTSC1/2AARAGsTOR:TargetofRapamycinMammalsHowisthismetabolicsignaltransduced?TORGlc-+++Rap+-WTtorGlucose-TORsignalingcontrolscellproliferationGlcGlc+RapGlc+AAGlcColtorEdUAfter24hoursglucosetreatmentRap:rapamycinAA:antimycinASpecificornonspecific?RapamycinGlycolysisinhibitorsMitochondriainhibitorsGeneralsignaling,transcriptionandtranslationSpecificsignalingeventmeristemactivation??Glucose-TORdoesnotaffectauxinandcytokininsignalingDR5::GFP-+-+-+-+IAAGlcGlc+RapGlc+AAtor-+-+-+-+ZtGlcGlc+RapGlc+AAtorTCS::GFPDR5::GFP,AuxinreporterTCS::GFP,CytokininreporterGlucose-TORsignalingspecificallycontrolsrootmeristemactivationRegulationofrootmeristemdoesnotrelyongeneraltranslationcontrolTranslation?Howdoesglucose-TORsignalingcontrolrootmeristemestablishment?NewFunctions?Transcription?mTORC1S6K4E-BPGrowthProliferation1050genes1318genesWTtorGlc-TORdirectsspecifictranscriptionalnetworks3daysundernoglucose,lowlight2hoursglucose(15mM)treatmentGlucoseGlycolysisTCAcycleETCATPCellcycle&DNAsynthesisGlycolysisTCAcycle&ETCRibosomebiogenesisProteinsynthesisAminoacidsynthesisNucleotidesynthesisRNAsynthesis&processingTFs&ChromatinmodulatorsSignalingTransportersCellwallsynthesisLigninsynthesisGlucosinolatesynthesisS-assimilationProteindegradationLipiddegradationAminoaciddegradationAutophagyCHOmetabolismBiodegradationofxenobioticsGluconeogeneseDevelopmentTFs&ChromatinmodulatorsStressTransportersMetabolictranscriptionnetworkTORkinaseP<0.01;Log2≥1GlucosetriggerscellcycleS-phasegeneexpressionchangeslog2ratio-3.003.0G1SG2MGlcG1GlcSGlcG2GlcM1082genes(Mengesetal.,2003)GlucosecontrolsG1toStransitionthroughE2Fa?RBRE2FsGeneexpressionCDC6,MCM3,MCM5ORC2,ORC6,PCNA1CDKACYCD3G1toStransition2695804Glc-TORup-regulatedE2FatargetgenesE2Fa_co-expressionE2Fa_up_regulatedgenes84E2Fa_binding_motifgenesE2FatargetedgenesGlucoseTORE2FaG1toStransition?FoldchangesE2Fa-HA-RapAMAtorWTE2FaP-T449S6K1TORS-phasegeneactivationbyE2FarequiresTORGlucoseTORE2FaG1toStransition32P-ATPE2FaTOR-++++Torin1--101001,000(nM)S-phasegeneactivationrequiresTOR-directlyphosphorylationofE2Fa14851624851624201420E2FAΔ1Δ2Δ3Δ48042032P-ATPE2Fa-6HISWTΔ1Δ2Δ3Δ4α-Flag+-+E2Fa-Flag-++IP:α-FlagW:α-TORW:α-FlagW:α-TORFoldchanges************************WTΔ1Δ2Δ3Δ4Glucose-TORsignalingdependsE2FaFoldchanges********+Glc/-GlcEdUWTe2faGlc-+-+WTe2fa3DAG+1DGlcGlucose-TORsignalinginrootmeristemactivationLeafSourcephotosynthesissucrose/glucoseRootSinkMeristemglycolysismitochondriaETC>OXP>ATPglucoseTORsignalingE2FaSphaseXiongandSheen,JBC,2012Xiongetal.,Nature,2013XiongandSheen,CellCycle,2013XiongandSheen,PlantPhysiology,2014Glucose-1-PUDP-GlucoseUGP1Glucose-6-PPGM3Fructose-6-PFructose-1,6-bis-PPFKs(At1g20950,At1g76550,At1g12000,At5g56630)Glycerinaldehyde-3-PAldolase

gene(At3g52930)Glycerate-1,3-bis-PGlycerate-3-PGlycerate-2-PPEPPyruvateGAPC1PGKIPGAM1,IPGAM2,ENO1,ENO2,ENOCPyruvatekinasegenes(At5g08570,At5g56350,At1g32440)Acetyl-CoACitrateIsocitrate2-oxoglutarateSuccinyl-CoASuccinateFumarateMalateOxaloacetateACO2,ACO3IDH1,At5g14590Succinyltransferase(At5g55070,At3g55410)Succinly-CoAligases(At5g23250,At5g08300,At2g20420)SDH1-1,SDH4FUM1mMDH1mMDH2SucroseGlucoseStarchGlycolysisTCAcycleTakehomemessagesSugarsignalingisverycomplexCrosstalkedwithhormonesSensedbydifferentmechanismsNewsignalingfunctionsofmetabolicenzymesSugarsignalnetwork

Itisjustthebeginning!!Baena-González,E.,Rolland,F.,Thevelein,J.M.

andSheen,J.2007.Acentralintegratoroftranscriptionnetworksinplantstressandenergysig