Iiproteni ze-DELLA ziyalondolozwaizilawuli zokukhulaezidlala indima ephambili ekuphuhliseni izityalo ekuphenduleni imiqondiso yangaphakathi nangaphandle. Njengabalawuli abashicilelweyo, ii-DELLA zibophelela kwizinto ezibhaliweyo (TFs) kunye ne-histone H2A ngokusebenzisa imimandla yazo ye-GRAS kwaye zigaywe ukuba zisebenze kubakhuthazi. Uphononongo lwakutsha nje lubonise ukuba ukuzinza kwe-DELLA kulawulwa emva kokuguqulelwa ngeendlela ezimbini: i-polyubiquitination eyenziwa yihomoni yesityalo.gibberellin, okukhokelela ekuthotyweni kwabo ngokukhawuleza, kunye nokudibanisa kunye ne-ubiquitin-like modifier (SUMO), eyandisa ukuqokelela kwabo. Ngaphezu koko, umsebenzi we-DELLA ulawulwa ngamandla ngeendlela ezimbini ezihlukeneyo ze-glycosylation: i-O-fucosylation iphucula ukusebenzisana kwe-DELLA-TF, kanti i-O-linked N-acetylglucosamine (O-GlcNAc) ukuguqulwa inqanda ukusebenzisana kwe-DELLA-TF. Nangona kunjalo, indima ye-DELLA phosphorylation ayicacanga njengoko izifundo zangaphambili zibonise iziphumo eziphikisanayo, abanye bacebisa ukuba i-phosphorylation ikhuthaza okanye icinezela ukuthotywa kwe-DELLA kunye nabanye bacebisa ukuba i-phosphorylation ayichaphazeli ukuzinza kwabo. Apha, sichonga iindawo ze-phosphorylation kwi-GA1-3 repressor (RGA), i-AtDELLA, ihlanjululwe kwi-Arabidopsis thaliana nge-mass spectrometry kwaye ibonisa ukuba i-phosphorylation yeepeptide ezimbini ze-RGA kwimimandla ye-PolyS kunye ne-PolyS / T iphucula umsebenzi we-RGA ngokukhuthaza ukubopha kwe-H2A kunye ne-RGA ukudibanisa nabakhuthazi abajoliswe kuyo. Ngokucacileyo, i-phosphorylation ayizange ichaphazele ukusebenzisana kwe-RGA-TF okanye ukuzinza kwe-RGA. Uphononongo lwethu lubonisa indlela yeemolekyuli apho i-phosphorylation ibangela umsebenzi we-DELLA.
Uhlalutyo lwethu lobunzima be-spectrometric lubonise ukuba zombini i-Pep1 kunye ne-Pep2 zine-phosphorylated kakhulu kwi-RGA kwi-GA-intsilelo ye-Ga1 yangasemva. Ukongeza kolu cwaningo, izifundo ze-phosphoproteomic ziye zatyhila i-Pep1 phosphorylation kwi-RGA, nangona indima yayo ingakafundwa53,54,55. Ngokwahlukileyo, i-Pep2 phosphorylation ayizange ichazwe ngaphambili kuba le peptide inokufunyanwa kuphela usebenzisa i-RGAGKG transgene. Nangona ukuguqulwa kwe-m1A, okuphelise i-phosphorylation ye-Pep1, kuphela ukunciphisa kancinci umsebenzi we-RGA kwi-planta, ibe nefuthe elongezelelweyo xa idibaniswa ne-m2A ekunciphiseni umsebenzi we-RGA (Umfanekiso owongezelelweyo 6). Okubalulekileyo, i-Pep1 phosphorylation yancitshiswa kakhulu kwi-GA-enhanced sly1 mutant xa kuthelekiswa ne-ga1, ebonisa ukuba i-GA ikhuthaza i-RGA dephosphorylation, ukunciphisa umsebenzi wayo. Indlela apho i-GA icinezela i-phosphorylation ye-RGA ifuna uphando olongezelelweyo. Enye into enokwenzeka kukuba oku kufezekiswa ngokulawulwa kweprotein kinase engaziwayo. Nangona uphando lubonise ukuba ukubonakaliswa kwe-CK1 protein kinase EL1 ilawulwa yi-GA kwi-rice41, iziphumo zethu zibonisa ukuba ukuguqulwa okuphezulu kwe-Arabidopsis EL1 homologue (AEL1-4) ayinciphisi i-RGA phosphorylation. Ngokuvumelana neziphumo zethu, uphando olutshanje lwe-phosphoproteomic olusebenzisa i-Arabidopsis AEL imigca yokugqithisa ngokugqithiseleyo kunye ne-ael triple mutant ayizange ibone nayiphi na iprotheni ye-DELLA njenge-substrates yale kinases56. Xa silungiselela umbhalo wesandla, kwaxelwa ukuba i-GSK3, i-gene encoding i-GSK3 / SHAGGY-like kinase kwingqolowa (Triticum aestivum), i-phosphorylate DELLA (Rht-B1b) 57, nangona i-phosphorylation ye-Rht-B1b nge-GSK3 ayizange iqinisekiswe kwi-planta. I-in vitro reactions enzymatic phambi kwe-GSK3 elandelwa luhlalutyo lwe-mass spectrometry lubonise iindawo ezintathu ze-phosphorylation ezibekwe phakathi kwe-DELLA kunye ne-GRAS domains ye-Rht-B1b (i-Fig. 3 eyoNgezelelweyo). I-Serine ukuya endaweni ye-alanine kuzo zonke iindawo ezintathu ze-phosphorylation zibangele ukuncipha komsebenzi we-Rht-B1b kwingqolowa ye-transgenic, ngokuhambelana neziphumo zethu zokuba ukutshintshwa kwe-alanine kwi-Pep2 RGA yehlise umsebenzi we-RGA. Nangona kunjalo, iimvavanyo zokuwohloka kweprotheyini ye-in vitro zibonise ngakumbi ukuba i-phosphorylation inokuzinzisa i-Rht-B1b57. Oku kuchasene neziphumo zethu ezibonisa ukuba ukutshintshwa kwe-alanine kwi-Pep2 RGA ayitshintshi ukuzinza kwayo kwi-planta. I-GSK3 kwingqolowa yi-ortholog ye-brassinosteroid-insensitive protein 2 (BIN2) kwi-Arabidopsis 57. I-BIN2 ngumlawuli ongalunganga wokubonakaliswa kwe-BR, kwaye i-BR isebenze indlela yayo yokubonisa ngokubangela i-BIN2 ukuthotywa kwe-58. Sibonise ukuba unyango lwe-BR alunciphisi i-RGA ukuzinza kwe-59 okanye i-phosphory2 ye-phosphory (i-ArabiSupplement) ecebisa ukuba i-RGA ayinakwenzeka ukuba ibe nephosphorylated yi-BIN2.
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Ixesha lokuposa: Apr-15-2025