Phosphorylation yana kunna mai kula da girma na DELLA a cikin Arabidopsis ta hanyar haɓaka alaƙar histone H2A da chromatin.

Sunadaran DELLA suna da matuƙar amfanimasu kula da ci gabanwaɗanda ke taka muhimmiyar rawa wajen sarrafa ci gaban tsirrai bisa ga alamun ciki da muhalli. DELLA tana aiki a matsayin mai kula da rubutu kuma ana ɗaukarta don kai hari ga masu haɓaka ta hanyar ɗaurewa ga abubuwan da ke haifar da rubutu (TFs) da histone H2A ta hanyar yankin GRAS ɗinta. Binciken da aka yi kwanan nan ya nuna cewa kwanciyar hankali na DELLA ana sarrafa shi ta hanyar fassara ta hanyoyi biyu: polyubiquitination da phytohormone gibberellin ke haifarwa, wanda ke haifar da raguwar sa cikin sauri, da haɗuwa da ƙananan masu gyara kamar ubiquitin (SUMO) don ƙara yawan tarinsa. Bugu da ƙari, ayyukan DELLA ana sarrafa su ta hanyar glycosylations guda biyu daban-daban: hulɗar DELLA-TF tana ƙaruwa ta hanyar O-fucosylation amma an hana ta ta hanyar gyaran O-linked N-acetylglucosamine (O-GlcNAc). Duk da haka, rawar da DELLA phosphorylation ta taka har yanzu ba a fayyace ba, kamar yadda binciken da ya gabata ya nuna sakamako masu karo da juna, tun daga waɗanda ke nuna cewa phosphorylation yana haɓaka ko rage lalacewar DELLA zuwa wasu waɗanda ke nuna cewa phosphorylation ba ya shafar kwanciyar hankalinsa. A nan, mun gano wuraren phosphorylation a cikin REPRESSORgaba 1-3(RGA, AtDELLA) an tsarkake shi daga Arabidopsis thaliana ta hanyar nazarin mass spectrometry kuma ya nuna cewa phosphorylation na peptides RGA guda biyu a yankunan PolyS da PolyS/T yana haɓaka haɗin H2A da haɓaka aikin RGA. Haɗin RGA da masu haɓaka manufa. Abin lura, phosphorylation ba ya shafar hulɗar RGA-TF ko kwanciyar hankali na RGA. Bincikenmu ya bayyana tsarin kwayoyin halitta wanda phosphorylation ke haifar da aikin DELLA.
Domin fayyace rawar da phosphorylation ke takawa wajen daidaita aikin DELLA, yana da matuƙar muhimmanci a gano wuraren phosphorylation na DELLA a cikin jiki da kuma yin nazarin aiki a cikin tsire-tsire. Ta hanyar tsarkakewar tsirrai da aka cire daga ciki, sannan aka binciko MS/MS, mun gano phosphosites da dama a cikin RGA. A ƙarƙashin yanayin ƙarancin GA, phosphorylation na RHA yana ƙaruwa, amma phosphorylation ba ya shafar kwanciyar hankalinsa. Abu mafi mahimmanci, gwaje-gwajen co-IP da ChIP-qPCR sun nuna cewa phosphorylation a yankin PolyS/T na RGA yana haɓaka hulɗarsa da H2A da alaƙarsa da masu haɓaka manufa, yana bayyana hanyar da phosphorylation ke haifar da aikin RGA.
Ana ɗaukar RGA don kai hari ga chromatin ta hanyar hulɗar subdomain na LHR1 tare da TF sannan ya haɗu da H2A ta yankin PolyS/T da subdomain na PFYRE, yana samar da hadaddun H2A-RGA-TF don daidaita RGA. Phosphorylation na Pep 2 a yankin PolyS/T tsakanin yankin DELLA da yankin GRAS ta hanyar kinase da ba a san ko waye ba yana haɓaka haɗin RGA-H2A. Protein ɗin rgam2A yana kawar da phosphorylation na RGA kuma yana ɗaukar wani tsari daban na furotin don tsoma baki ga haɗin H2A. Wannan yana haifar da rashin daidaituwar hulɗar TF-rgam2A na ɗan lokaci da rabuwar rgam2A daga chromatin da aka yi niyya. Wannan adadi yana nuna kawai matsin lamba na rubutu wanda RGA ke jagoranta. Ana iya bayyana irin wannan tsari don kunna rubutun da RGA ke jagoranta, sai dai cewa hadaddun H2A-RGA-TF zai haɓaka rubutun kwayoyin halitta da kuma rage phosphorylation na rgam2A zai rage rubutun. An gyara hoto daga Huang et al.21.
An yi nazarin dukkan bayanai na adadi ta amfani da Excel, kuma an tantance manyan bambance-bambance ta amfani da gwajin t na ɗalibi. Ba a yi amfani da hanyoyin ƙididdiga don tantance girman samfurin ba tukuna. Ba a cire bayanai daga binciken ba; gwajin ba a yi shi ba bisa ka'ida ba; masu binciken ba su makance ga rarraba bayanai yayin gwajin da kuma kimanta sakamakon ba. An nuna girman samfurin a cikin tatsuniyar adadi da fayil ɗin bayanan tushe.
Don ƙarin bayani game da tsarin binciken, duba Takaitaccen Rahoton Fayil na Halitta wanda ke da alaƙa da wannan labarin.
An ba da gudummawar bayanai na proteomics na Mass spectrometry ga ƙungiyar ProteomeXchange ta hanyar ma'ajiyar abokin hulɗar PRIDE66 tare da mai gano bayanai na PXD046004. Duk sauran bayanan da aka samu a lokacin wannan binciken an gabatar da su a cikin Ƙarin Bayani, Fayilolin Bayanai na Ƙarin Bayani, da Fayilolin Bayanai na Raw. An bayar da bayanai na tushe don wannan labarin.