I-Phosphorylation ivuselela umlawuli wokukhula oyintloko u-DELLA kwi-Arabidopsis ngokukhuthaza umbutho we-histone H2A kunye ne-chromatin.

Iiproteni ze-DELLA ziyinkosi egciniweyoizilawuli zokukhulaezidlala indima ephambili ekulawuleni uphuhliso lwezityalo ekuphenduleni kwiimpawu zangaphakathi kunye nokusingqongileyo. I-DELLA isebenza njengomlawuli wokubhaliweyo kwaye igaywe ukuba ijolise kubakhuthazi ngokuzibophelela kwizinto ezibhaliweyo (TFs) kunye ne-histone H2A ngokusebenzisa isizinda sayo se-GRAS. Uphononongo olutshanje lubonise ukuba ukuzinza kwe-DELLA kulawulwa emva kokuguqulelwa ngeendlela ezimbini: i-polyubiquitination eyenziwa yi-phytohormone gibberellin, ekhokelela ekuthotyweni kwayo ngokukhawuleza, kunye nokudibanisa kwezinto ezincinci ezifana ne-ubiquitin-like modifiers (SUMO) ukwandisa ukuqokelela kwayo. Ukongezelela, umsebenzi we-DELLA ulawulwa ngamandla ngama-glycosylations amabini ahlukeneyo: ukusebenzisana kwe-DELLA-TF kuphuculwe yi-O-fucosylation kodwa kuvinjelwe yi-O-linked N-acetylglucosamine (O-GlcNAc) ukuguqulwa. Nangona kunjalo, indima ye-DELLA phosphorylation ihlala ingacacanga, njengoko izifundo zangaphambili zibonise iziphumo eziphikisanayo, ukusuka kwezo zibonisa ukuba i-phosphorylation ikhuthaza okanye iyanciphisa ukuthotywa kwe-DELLA kwabanye ebonisa ukuba i-phosphorylation ayichaphazeli ukuzinza kwayo. Apha, sichonga iindawo ze-phosphorylation kwi-REPRESSORiGa1-3(i-RGA, i-AtDELLA) ihlanjululwe kwi-Arabidopsis thaliana ngokuhlalutya kwe-mass spectrometry kwaye ibonisa ukuba i-phosphorylation yeepeptide ezimbini ze-RGA kwimimandla ye-PolyS kunye ne-PolyS / T ikhuthaza ukubopha kwe-H2A kunye nokuphucula umsebenzi we-RGA. Umanyano lwe-RGA nabakhuthazi ekujoliswe kubo. Ngokucacileyo, i-phosphorylation ayichaphazeli ukusebenzisana kwe-RGA-TF okanye ukuzinza kwe-RGA. Uphononongo lwethu lubonisa indlela yeemolekyuli apho i-phosphorylation ibangela umsebenzi we-DELLA.
Ukucacisa indima ye-phosphorylation ekulawuleni umsebenzi we-DELLA, kubalulekile ukuchonga iindawo ze-DELLA phosphorylation kwi-vivo kwaye wenze uhlalutyo olusebenzayo kwizityalo. Ngokucocwa kobudlelwane bezityalo ezilandelwa luhlalutyo lwe-MS/MS, sichonge iiphosphosites ezininzi kwi-RGA. Ngaphantsi kweemeko zokusilela kwe-GA, i-RHA phosphorylation iyanda, kodwa i-phosphorylation ayichaphazeli ukuzinza kwayo. Okubalulekileyo, i-co-IP kunye ne-ChIP-qPCR i-assays ibonise ukuba i-phosphorylation kwindawo ye-PolyS / T ye-RGA ikhuthaza ukusebenzisana kwayo kunye ne-H2A kunye nokudibanisa nabakhuthazi abajoliswe kuyo, ityhila indlela apho i-phosphorylation ibangela umsebenzi we-RGA.
I-RGA igaywe ukuba ijolise kwi-chromatin ngokusebenzisa i-LHR1 subdomain kunye ne-TF kwaye ibophe kwi-H2A ngengingqi yayo ye-PolyS / T kunye ne-PFYRE subdomain, eyenza i-H2A-RGA-TF eyinkimbinkimbi yokuzinzisa i-RGA. I-Phosphorylation ye-Pep 2 kummandla we-PolyS/T phakathi kwesizinda se-DELLA kunye nesizinda se-GRAS nge-kinase engaziwayo iphucula i-RGA-H2A yokubopha. Iprotheni eguquguqukayo ye-rgam2A ichitha i-phosphorylation ye-RGA kwaye ithatha i-protein ye-protein ehlukeneyo ukuphazamisa ukubopha kwe-H2A. Oku kubangela ukungazinzi kwentsebenziswano ye-TF-rgam2A yexesha elifutshane kunye nokuchithwa kwe-rgam2A kwi-chromatin ekujoliswe kuyo. Eli nani libonisa kuphela i-RGA-mediated transcriptional repression. Ipateni efanayo inokuchazwa kwi-RGA-mediated transcriptional activation, ngaphandle kokuba i-H2A-RGA-TF complex ingakhuthaza ushicilelo lwejene ekujoliswe kulo kunye ne-dephosphorylation ye-rgam2A iya kunciphisa ushicilelo. Umzobo ulungiswe ukusuka ku-Huang et al.21.
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Ngolwazi oluthe vetshe malunga noyilo lophononongo, bona i-Natural Portfolio Report Abstract ehambelana neli nqaku.
Idatha ye-mass spectrometry proteomics ifakwe kwi-ProteomeXchange consortium ngendawo yokugcina iqabane le-PRIDE66 kunye nesazisi sedatha ye-PXD046004. Zonke ezinye iinkcukacha ezifunyenwe ngexesha lolu phononongo zinikezelwe kwiiNkcukacha ezoNgezelelweyo, iiFayile zeDatha ezoNgezelelweyo, kunye neeFayile zeDatha eRaw. Idatha yomthombo inikwe eli nqaku.