I-Phosphorylation ivuselela i-master growth regulator DELLA kwi-Arabidopsis ngokukhuthaza ukudibana kwe-histone H2A ne-chromatin.

Iiproteni zeDella zilungile kakhuluabalawuli bokukhulaezidlala indima ephambili ekulawuleni uphuhliso lwezityalo ngokuphendula kwiimpawu zangaphakathi nezendalo. I-DELLA isebenza njengomlawuli we-transcriptional kwaye iqeshwa ukuba ijolise kubakhuthazi ngokubophelela kwizinto ze-transcription (TFs) kunye ne-histone H2A ngokusebenzisa indawo yayo ye-GRAS. Izifundo zakutshanje zibonise ukuba uzinzo lwe-DELLA lulawulwa emva koguqulelo ngeendlela ezimbini: i-polyubiquitination ebangelwa yi-phytohormone gibberellin, ekhokelela ekuwohlokeni kwayo ngokukhawuleza, kunye nokudibanisa ii-small ubiquitin-like modifiers (SUMO) ukwandisa ukuqokelelana kwayo. Ukongeza, umsebenzi we-DELLA ulawulwa ngokuguquguqukayo zii-glycosylations ezimbini ezahlukeneyo: ukusebenzisana kwe-DELLA-TF kuphuculwe yi-O-fucosylation kodwa kuthintelwe kukuguqulwa kwe-O-linked N-acetylglucosamine (O-GlcNAc). Nangona kunjalo, indima ye-DELLA phosphorylation ayikacaci, njengoko izifundo zangaphambili zibonise iziphumo ezingqubanayo, ukusuka kwezo zibonisa ukuba i-phosphorylation ikhuthaza okanye inciphisa ukonakala kwe-DELLA ukuya kwezinye ezibonisa ukuba i-phosphorylation ayichaphazeli uzinzo lwayo. Apha, sichonga iindawo ze-phosphorylation kwi-REPRESSORiga1-3(RGA, AtDELLA) ihlanjululwe kwi-Arabidopsis thaliana ngohlalutyo lwe-mass spectrometry kwaye ibonisa ukuba i-phosphorylation yee-peptides ezimbini ze-RGA kwiindawo ze-PolyS kunye ne-PolyS/T ikhuthaza umsebenzi we-H2A wokubopha nokuphucula i-RGA. Ukudibana kwe-RGA kunye nabakhuthazi abajoliswe kuyo. Okuphawulekayo kukuba, i-phosphorylation ayichaphazeli ukusebenzisana kwe-RGA-TF okanye ukuzinza kwe-RGA. Uphononongo lwethu lubonisa indlela i-molecule esebenza ngayo i-phosphorylation ebangela umsebenzi we-DELLA.
Ukuze kucaciswe indima ye-phosphorylation ekulawuleni umsebenzi we-DELLA, kubalulekile ukuchonga iindawo ze-DELLA phosphorylation in vivo kunye nokwenza uhlalutyo olusebenzayo kwizityalo. Ngokucocwa kwe-affinity kwezicatshulwa zezityalo kulandelwa luhlalutyo lwe-MS/MS, sichonge ii-phosphosite ezininzi kwi-RGA. Phantsi kweemeko zokungabikho kwe-GA, i-RHA phosphorylation iyanda, kodwa i-phosphorylation ayichaphazeli uzinzo lwayo. Okubalulekileyo, uvavanyo lwe-co-IP kunye ne-ChIP-qPCR lutyhile ukuba i-phosphorylation kummandla we-PolyS/T we-RGA ikhuthaza ukusebenzisana kwayo ne-H2A kunye nokudibana kwayo nabakhuthazi abajoliswe kuyo, nto leyo ebonisa indlela i-phosphorylation ebangela ngayo umsebenzi we-RGA.
I-RGA iqeshwa ukuze ijolise kwi-chromatin ngokusebenzisana kwe-subdomain ye-LHR1 kunye ne-TF ize ibophelele kwi-H2A ngokusebenzisa ummandla wayo we-PolyS/T kunye ne-subdomain ye-PFYRE, yenze i-H2A-RGA-TF complex ukuze kuzinziswe i-RGA. I-Phosphorylation ye-Pep 2 kummandla we-PolyS/T phakathi kwe-domain ye-DELLA kunye ne-GRAS domain yi-kinase engaziwayo iphucula ukubopha kwe-RGA-H2A. Iprotheyini ye-rgam2A mutant isusa i-RGA phosphorylation kwaye yamkela ukwakheka kweprotheyini eyahlukileyo ukuphazamisana nokubopha kwe-H2A. Oku kuphumela ekungazinziseni kokunxibelelana kwe-TF-rgam2A okwethutyana kunye nokwahlukana kwe-rgam2A kwi-chromatin ekujoliswe kuyo. Lo mfanekiso ubonisa kuphela ucinezelo lwe-transcriptional oluxhaswa yi-RGA. Ipatheni efanayo ingachazwa ekusebenzeni kwe-transcriptional okuxhaswe yi-RGA, ngaphandle kokuba i-complex ye-H2A-RGA-TF iya kukhuthaza ukubhalwa kwe-gene ekujoliswe kuyo kunye nokususwa kwe-phosphorylation ye-rgam2A kuya kunciphisa ukubhalwa. Umfanekiso uguqulwe kuHuang et al.21.
Zonke iinkcukacha zobungakanani zihlalutywe ngokwezibalo kusetyenziswa i-Excel, kwaye umahluko obalulekileyo uchongiwe kusetyenziswa uvavanyo lwe-t lwabafundi. Akukho ndlela zezibalo ezisetyenzisiweyo ukumisela ubungakanani besampulu kwangaphambili. Akukho datha ikhutshiweyo kuhlalutyo; uvavanyo aluzange lube nocwangco; abaphandi babengaboni kakuhle ukusasazwa kwedatha ngexesha lovavanyo kunye novavanyo lweziphumo. Ubungakanani besampulu buboniswe kwintsomi yomfanekiso kunye nefayile yedatha yomthombo.
Ukuze ufumane ulwazi oluthe kratya malunga noyilo lophando, jonga i-Natural Portfolio Report Abstract enxulumene neli nqaku.
Idatha ye-Mass spectrometry proteomics inikelwe kwi-ProteomeXchange consortium ngokusebenzisa indawo yokugcina amaqabane ye-PRIDE66 ene-dataset identifier PXD046004. Zonke ezinye iinkcukacha ezifunyenweyo ngexesha lolu phononongo ziboniswe kwi-Supplementary Information, ii-Supplementary Data Files, kunye nee-Raw Data Files. Idatha yomthombo inikwe kweli nqaku.