Fosifora itera imbaraga zo gukura gukura DELLA, guteza imbere histone H2A ihuza chromatine muri Arabidopsis.

Intungamubiri za DELLA zabitsweabashinzwe iteramberebigira uruhare runini mugutezimbere ibimera hasubizwa ibimenyetso byimbere ninyuma. Nkabashinzwe kugenzura inyandiko-mvugo, DELLAs ihuza ibintu byandikirwa (TFs) na histone H2A ikoresheje domaine zabo za GRAS kandi bashakirwa gukora kubamamaza. Ubushakashatsi buherutse gukorwa bwerekanye ko umutekano wa DELLA ugengwa nyuma yubusobanuro nuburyo bubiri: polyubiquitination iterwa na hormone yibihingwagibberellin, biganisha ku kwangirika kwabo vuba, no guhuza hamwe na ubiquitin ntoya isa na modifier (SUMO), byongera kwirundanya kwabo. Byongeye kandi, ibikorwa bya DELLA bigengwa nuburyo bubiri butandukanye bwa glycosylation: O-fucosylation yongerera imikoranire ya DELLA-TF, mugihe O-ihuza N-acetylglucosamine (O-GlcNAc) ihindura imikoranire ya DELLA-TF. Icyakora, uruhare rwa fosifora ya DELLA ntirusobanutse neza kuko ubushakashatsi bwibanze bwerekanye ibisubizo bivuguruzanya, aho bamwe bavuga ko fosifora itera cyangwa igahagarika iyangirika rya DELLA abandi bakavuga ko fosifora itagira ingaruka ku gutuza kwabo. Hano, tumenye ibibanza bya fosifora muri repressor ya GA1-3 (RGA), AtDELLA, byahanaguwe na Arabidopsis thaliana na mass spectrometrie kandi twerekana ko fosifora ya peptide ebyiri za RGA mukarere ka PolyS na PolyS / T byongera ibikorwa bya RGA mugutezimbere H2A guhuza hamwe na RGA hamwe nabateza imbere intego. Ikigaragara ni uko fosifora itagize ingaruka ku mikoranire ya RGA-TF cyangwa ituze rya RGA. Ubushakashatsi bwacu bugaragaza uburyo bwa molekuline ikoreshwa na fosifora itera ibikorwa bya DELLA.
Isesengura rusange ryakozwe ryerekanye ko Pep1 na Pep2 byombi byari fosifori cyane muri RGA inyuma ya GA yabuze GA1. Usibye ubu bushakashatsi, ubushakashatsi bwa fosifore bwerekanye kandi Pep1 fosifora muri RGA, nubwo uruhare rwayo rutarakorwa 53,54,55. Ibinyuranye, Pep2 ya fosifora ntabwo yigeze isobanurwa kuva iyi peptide yashoboraga kumenyekana hakoreshejwe transgene ya RGAGKG. Nubwo ihinduka rya m1A, ryakuyeho fosifora ya Pep1, ryagabanije gato ibikorwa bya RGA muri planta, ryagize ingaruka zinyongera iyo rihujwe na m2A mukugabanya ibikorwa bya RGA (Ishusho yinyongera 6). Icy'ingenzi, Pep1 fosifora yagabanutse cyane muri GA yongerewe imbaraga ya sly1 ugereranije na ga1, byerekana ko GA iteza imbere RGA dephosifora, igabanya ibikorwa byayo. Uburyo GA ihagarika fosifora ya RGA bisaba ko hakorwa iperereza rindi. Ikintu kimwe gishoboka nuko ibyo bigerwaho hifashishijwe amabwiriza ya protein kinase itazwi. Nubwo ubushakashatsi bwerekanye ko imvugo ya CK1 protein kinase EL1 itagengwa na GA mu muceri41, ibisubizo byacu byerekana ko ihinduka ry’imiterere ihanitse ya Arabidopsis EL1 homologue (AEL1-4) itagabanya fosifori ya RGA. Mu buryo buhuye n'ibisubizo byacu, ubushakashatsi bwa fosifore-protomique iherutse gukoresha imirongo ya Arabidopsis AEL ikabije kandi na ael triple mutant ntiyigeze igaragaza poroteyine zose za DELLA nk'intungamubiri za kinase56. Igihe twateguraga ibyandikishijwe intoki, byavuzwe ko GSK3, gene irimo GSK3 / SHAGGY isa na kinase mu ngano (Triticum aestivum), ishobora gutera fosifori DELLA (Rht-B1b) 57, nubwo fosifora ya Rht-B1b na GSK3 itaremezwa muri planta. Muri vitro enzymatique ya reaction imbere ya GSK3 ikurikirwa nisesengura rusange rya sprometrike yerekanaga ibibanza bitatu bya fosifora biri hagati ya DELLA na GRAS ya Rht-B1b (Ishusho yinyongera). Serine kuri alanine isimburwa kuri fosifora zose uko ari eshatu zatumye ibikorwa bya Rht-B1b bigabanuka mu ngano ya transgenji, bihuye nibyo twabonye ko insimburangingo ya alanine muri Pep2 RGA yagabanije ibikorwa bya RGA. Nyamara, muri vitro protein yangirika yerekana ko fosifora nayo ishobora guhagarika Rht-B1b57. Ibi bitandukanye nibisubizo byacu byerekana ko insimburangingo ya alanine muri Pep2 RGA idahindura ituze muri planta. GSK3 mu ngano ni orthologique ya brassinosteroid-idafite insimburangingo ya poroteyine 2 (BIN2) muri Arabidopsis 57. BIN2 ni igenzura ribi ryerekana ibimenyetso bya BR, kandi BR itangiza inzira yayo yerekana ibimenyetso itera kwangirika kwa BIN2 58. Twerekanye ko ubuvuzi bwa BR butagabanya umutekano wa RGA 59 cyangwa ngo fosifora ikorwe na Arabidopsis (byerekana ko B2).
Umubare wimibare yose wasesenguwe hifashishijwe Excel, kandi itandukaniro rikomeye ryagaragaye ukoresheje t-test yabanyeshuri. Nta buryo bwibarurishamibare bwakoreshejwe mbere yo kumenya ingano yicyitegererezo. Nta makuru yakuwe mu isesengura; ubushakashatsi ntabwo bwatoranijwe; n'abashakashatsi bari bazi kugenwa mugihe cyo kugerageza no gusuzuma ibisubizo. Ingano ntangarugero itangwa mumigani yimibare no mumadosiye yibanze.