Phosphorylation yana kunna mai sarrafa girma mai sarrafa DELLA a cikin Arabidopsis ta haɓaka haɗin gwiwar histone H2A tare da chromatin.

Sunadaran DELLA an kiyaye sumasu kula da girmawanda ke taka muhimmiyar rawa wajen sarrafa ci gaban shuka don amsa alamun ciki da muhalli. DELLA tana aiki a matsayin mai tsara kwafi kuma ana ɗaukarta don niyya ga masu tallata ta hanyar ɗaure abubuwan rubutu (TFs) da histone H2A ta hanyar yankin GRAS. Binciken na baya-bayan nan ya nuna cewa an daidaita daidaiton DELLA bayan fassarar ta hanyoyi guda biyu: polyubiquitination wanda phytohormone gibberellin ke haifar da shi, wanda ke haifar da raguwa da sauri, da haɗuwa da ƙananan gyare-gyare masu kama da ubiquitin (SUMO) don ƙara yawan tarawa. Bugu da ƙari, ayyukan DELLA ana daidaita su ta hanyar glycosylations daban-daban guda biyu: hulɗar DELLA-TF yana haɓaka ta O-fucosylation amma an hana shi ta hanyar O-linked N-acetylglucosamine (O-GlcNAc). Duk da haka, aikin DELLA phosphorylation ya kasance ba a sani ba, kamar yadda binciken da aka yi a baya ya nuna sakamako masu cin karo da juna, daga wadanda ke nuna cewa phosphorylation yana inganta ko rage lalata DELLA ga wasu yana nuna cewa phosphorylation ba ya shafar kwanciyar hankali. Anan, mun gano rukunin phosphorylation a cikin REPRESSORgaba 1-3(RGA, AtDELLA) wanda aka tsarkake daga Arabidopsis thaliana ta hanyar bincike mai yawa kuma ya nuna cewa phosphorylation na peptides RGA guda biyu a yankunan PolyS da PolyS/T yana haɓaka haɗin H2A da haɓaka ayyukan RGA. Ƙungiyar RGA tare da masu tallata manufa. Musamman, phosphorylation baya shafar hulɗar RGA-TF ko kwanciyar hankali na RGA. Bincikenmu yana bayyana tsarin kwayoyin halitta wanda phosphorylation ke haifar da ayyukan DELLA.
Don bayyana rawar phosphorylation a cikin daidaita aikin DELLA, yana da mahimmanci a gano wuraren DELLA phosphorylation a cikin vivo da yin nazarin aiki a cikin tsire-tsire. Ta hanyar tsarkakewa na tsantsar tsire-tsire da MS/MS bincike ya biyo baya, mun gano abubuwan phosphosites da yawa a cikin RGA. A ƙarƙashin yanayin ƙarancin GA, RHA phosphorylation yana ƙaruwa, amma phosphorylation ba ya shafar kwanciyar hankali. Mahimmanci, ƙididdigar co-IP da ChIP-qPCR sun bayyana cewa phosphorylation a yankin PolyS / T na RGA yana inganta hulɗar ta tare da H2A da haɗin gwiwa tare da masu tallata manufa, yana nuna hanyar da phosphorylation ke haifar da aikin RGA.
Ana ɗaukar RGA don ƙaddamar da chromatin ta hanyar hulɗar yanki na LHR1 tare da TF sannan ya ɗaure zuwa H2A ta yankin PolyS/T da PFYRE reshen yanki, yana samar da hadaddun H2A-RGA-TF don daidaita RGA. Phosphorylation na Pep 2 a cikin yankin PolyS/T tsakanin yankin DELLA da yankin GRAS ta wani kinase da ba a bayyana ba yana haɓaka ɗaurin RGA-H2A. Mutant protein na rgam2A yana soke RGA phosphorylation kuma yana ɗaukar nau'in nau'in furotin na daban don tsoma baki tare da ɗaurin H2A. Wannan yana haifar da rashin kwanciyar hankali na hulɗar TF-rgam2A na wucin gadi da rarrabuwa na rgam2A daga chromatin da aka yi niyya. Wannan adadi yana kwatanta danniya mai tsaka-tsaki na RGA kawai. Ana iya siffanta irin wannan tsari don kunna rubutu na tsaka-tsaki na RGA, sai dai cewa hadaddun H2A-RGA-TF zai inganta fassarar kwayar halittar manufa da dephosphorylation na rgam2A zai rage rubutun. Hoton da aka gyara daga Huang et al.21.
Dukkanin bayanan ƙididdiga an yi nazarin su ta hanyar amfani da Excel, kuma an ƙaddara bambance-bambance masu mahimmanci ta amfani da gwajin Student's t. Ba a yi amfani da hanyoyin ƙididdiga don tantance girman samfurin da farko ba. Babu bayanai da aka cire daga bincike; gwajin ba a kayyade ba; masu binciken ba su makantar da rarraba bayanai yayin gwajin da kuma kimanta sakamakon. Ana nuna girman samfurin a cikin almara na adadi da fayil ɗin bayanan tushe.
Don ƙarin bayani game da ƙirar binciken, duba Abstract Rahoton Fayil na Halitta mai alaƙa da wannan labarin.
An ba da gudummawar bayanan ƙididdiga masu ƙima ga ƙungiyar ProteomeXchange ta wurin ma'ajin abokin tarayya na PRIDE66 tare da mai gano saitin bayanai PXD046004. Duk sauran bayanan da aka samu yayin wannan binciken ana gabatar da su a cikin Ƙarin Bayani, Ƙarin Fayilolin Bayanai, da Fayilolin Bayanan Raw. An bayar da bayanan tushen don wannan labarin.