Amaprotheni e-DELLA agcinwa kahleabalawuli bokukhulaezidlala indima ebalulekile ekulawuleni intuthuko yezitshalo ekuphenduleni izinkomba zangaphakathi nezendawo ezungezile. I-DELLA isebenza njengomlawuli wokubhala futhi iqashwa ukuba iqondise abagqugquzeli ngokuzibopha ku-transcription factors (TFs) kanye ne-histone H2A ngokusebenzisa isizinda sayo se-GRAS. Izifundo zakamuva zibonise ukuthi ukuzinza kwe-DELLA kulawulwa ngemuva kokuhumusha ngezindlela ezimbili: i-polyubiquitination ebangelwa yi-phytohormone gibberellin, okuholela ekuwohlokeni kwayo okusheshayo, kanye nokuhlanganiswa kwama-modifiers amancane afana ne-ubiquitin (SUMO) ukwandisa ukuqongelela kwayo. Ngaphezu kwalokho, umsebenzi we-DELLA ulawulwa ngokuguquguqukayo yi-glycosylations ezimbili ezahlukene: ukusebenzisana kwe-DELLA-TF kukhuliswa yi-O-fucosylation kodwa kuvinjelwe ukuguqulwa kwe-O-linked N-acetylglucosamine (O-GlcNAc). Kodwa-ke, indima ye-DELLA phosphorylation ayikacaci, njengoba izifundo zangaphambilini zibonise imiphumela ephikisanayo, kusukela kulezo ezibonisa ukuthi i-phosphorylation ikhuthaza noma inciphisa ukonakala kwe-DELLA kuya kwezinye ezibonisa ukuthi i-phosphorylation ayithinti ukuzinza kwayo. Lapha, sithola izindawo ze-phosphorylation ku-REPRESSORiga1-3(RGA, AtDELLA) ihlanzwe ku-Arabidopsis thaliana ngokuhlaziywa kwe-mass spectrometry futhi ikhombisa ukuthi i-phosphorylation yama-peptide amabili e-RGA ezifundeni ze-PolyS kanye ne-PolyS/T ikhuthaza umsebenzi we-H2A wokubopha nokuthuthukisa i-RGA. Ukuhlanganiswa kwe-RGA nabagqugquzeli abaqondiwe. Okuphawulekayo ukuthi i-phosphorylation ayithinti ukusebenzisana kwe-RGA-TF noma ukuzinza kwe-RGA. Ucwaningo lwethu lwembula indlela ama-molecule asebenzisa ngayo i-phosphorylation ekubangela umsebenzi we-DELLA.
Ukuze kucaciswe indima ye-phosphorylation ekulawuleni umsebenzi we-DELLA, kubalulekile ukuhlonza izindawo ze-phosphorylation ze-DELLA in vivo nokwenza ukuhlaziywa kokusebenza ezitshalweni. Ngokuhlanzwa kwe-affinity kwezingcezu zezitshalo kulandelwe ukuhlaziywa kwe-MS/MS, sithole ama-phosphosite amaningana ku-RGA. Ngaphansi kwezimo zokuntuleka kwe-GA, i-phosphorylation ye-RHA iyanda, kodwa i-phosphorylation ayithinti ukuzinza kwayo. Okubalulekile, ukuhlolwa kwe-co-IP kanye ne-ChIP-qPCR kwembule ukuthi i-phosphorylation esifundeni se-PolyS/T se-RGA ikhuthaza ukusebenzisana kwayo ne-H2A kanye nokuhlangana kwayo nabagqugquzeli abaqondiwe, okwembula indlela i-phosphorylation ebangela ngayo umsebenzi we-RGA.
I-RGA iqashwa ukuze iqondise i-chromatin ngokusebenzisana kwesizinda esincane se-LHR1 ne-TF bese ibopha ku-H2A ngokusebenzisa isifunda sayo se-PolyS/T kanye nesizinda esincane se-PFYRE, yakha i-H2A-RGA-TF complex ukuze kuzinziswe i-RGA. I-Phosphorylation ye-Pep 2 esifundeni se-PolyS/T phakathi kwesizinda se-DELLA kanye nesizinda se-GRAS yi-kinase engaziwa ithuthukisa ukubopha kwe-RGA-H2A. Iphrotheni ye-rgam2A mutant iqeda i-RGA phosphorylation futhi yamukela ukwakheka kwephrotheni okuhlukile ukuze iphazamise ukubopha kwe-H2A. Lokhu kuphumela ekungazinzileni kokuxhumana kwe-TF-rgam2A okwesikhashana kanye nokuhlukaniswa kwe-rgam2A kusuka ku-chromatin eqondiwe. Lesi sibalo sibonisa kuphela ukucindezelwa kokubhala okuqondiswe yi-RGA. Iphethini efanayo ingachazwa ekusebenzeni kokubhala okuqondiswe yi-RGA, ngaphandle kokuthi i-H2A-RGA-TF complex izokhuthaza ukubhalwa kwezakhi zofuzo okuqondiwe kanye nokususwa kwe-rgam2A okuzonciphisa ukubhalwa. Isithombe sishintshiwe kusuka ku-Huang et al.21.
Yonke idatha yokulinganisa ihlaziywe ngokwezibalo kusetshenziswa i-Excel, futhi umehluko omkhulu watholakala kusetshenziswa isivivinyo sika-t somfundi. Azikho izindlela zezibalo ezisetshenziswe ukunquma usayizi wesampula ekuqaleni. Akukho datha ekhishwe ekuhlaziyweni; isivivinyo asizange sihlelwe ngokungahleliwe; abacwaningi babengaboni ukusatshalaliswa kwedatha ngesikhathi sokuhlolwa kanye nokuhlolwa kwemiphumela. Usayizi wesampula ukhonjiswe ku-figure legend kanye nefayela ledatha yomthombo.
Ukuze uthole ulwazi olwengeziwe mayelana nomklamo wocwaningo, bheka i-Natural Portfolio Report Abstract ehlobene nalesi sihloko.
Idatha ye-proteomics ye-mass spectrometry inikelwe ku-ProteomeXchange consortium ngokusebenzisa indawo yokugcina umlingani we-PRIDE66 ene-dataset identifier PXD046004. Yonke eminye imininingwane etholwe phakathi nalolu cwaningo inikezwe ku-Supplementary Information, Supplementary Data Files, kanye ne-Raw Data Files. Idatha yomthombo inikezwe kulesi sihloko.
Isikhathi sokuthunyelwe: Novemba-08-2024