Phosphorylation inomutsa tenzi kukura regulator DELLA, inosimudzira histone H2A inosunga kune chromatin muArabidopsis.

DELLA mapuroteni anochengetedzwakukura regulatorsiyo inoita basa guru mukukura kwezvirimwa mukupindura zviratidzo zvemukati nekunze. Sevamiriri vezvinyorwa, DELLAs inosunga kune zvinyorwa zvinyorwa (TFs) uye histone H2A kuburikidza nematunhu avo eGRAS uye vanopinzwa basa kune vanosimudzira. Zvidzidzo zvenguva pfupi yapfuura zvakaratidza kuti kugadzikana kweDELLA kunodzorwa mushure mekushandura nenzira mbiri: polyubiquitination inokonzerwa nehomoni yechirimwa.gibberellin, iyo inotungamira mukuparara kwavo nekukurumidza, uye kusanganisa nediki ubiquitin-senge modifier (SUMO), iyo inowedzera kuunganidza kwavo. Uyezve, chiitiko cheDELLA chinodzorwa zvine simba nemaitiro maviri akasiyana eglycosylation: O-fucosylation inosimudzira DELLA-TF kudyidzana, nepo O-yakabatana N-acetylglucosamine (O-GlcNAc) shanduko inotadzisa DELLA-TF kudyidzana. Nekudaro, basa reDELLA phosphorylation harina kujeka sezvo zvidzidzo zvekare zvakaratidza mhedzisiro inopokana, vamwe vachiratidza kuti phosphorylation inosimudzira kana kudzvinyirira DELLA kuderedzwa uye vamwe vachiratidza kuti phosphorylation haikanganisi kugadzikana kwavo. Pano, isu tinocherekedza phosphorylation nzvimbo muGA1-3 repressor (RGA), AtDELLA, yakacheneswa kubva kuArabidopsis thaliana nehukuru spectrometry uye inoratidza kuti phosphorylation yemaviri eRGA peptides munzvimbo dzePolyS nePolyS/T inosimudzira RGA chiitiko nekusimudzira H2A kusunga uye RGA kushamwaridzana nevanosimudzira vanosimudzira. Zvikuru, phosphorylation haina kukanganisa RGA-TF kuwirirana kana RGA kugadzikana. Chidzidzo chedu chinoburitsa ma molecular maitiro ayo phosphorylation induces DELLA chiitiko.
Yedu yakawanda spectrometric ongororo yakaratidza kuti ese Pep1 nePep2 aive akanyanya phosphorylated muRGA muGA-inoshomeka Ga1 kumashure. Pamusoro pechidzidzo ichi, zvidzidzo zvephosphoproteomic zvakaratidzawo Pep1 phosphorylation muRGA, kunyange zvazvo basa rayo risati radzidzwa53,54,55. Kusiyana neizvi, Pep2 phosphorylation haisati yambotsanangurwa sezvo peptide iyi yaigona kungoonekwa uchishandisa RGAGKG transgene. Kunyangwe iyo m1A mutation, iyo yakabvisa Pep1 phosphorylation, yakangodzikisa zvishoma RGA chiitiko muplanta, yaive nekuwedzera mhedzisiro kana yasanganiswa nem2A mukudzikisa RGA chiitiko (Supplementary Figure 6). Zvakakosha, Pep1 phosphorylation yakaderedzwa zvakanyanya muGA-enhanced sly1 mutant ichienzaniswa nega1, zvichiratidza kuti GA inokurudzira RGA dephosphorylation, kuderedza basa rayo. Iyo nzira iyo GA inodzvanya RGA phosphorylation inoda kumwe kuferefetwa. Imwe mukana ndeyokuti izvi zvinowanikwa kuburikidza nekugadzirisa kweprotein kinase isingazivikanwi. Kunyange zvazvo zvidzidzo zvakaratidza kuti kutaura kweCK1 protein kinase EL1 yakaderedzwa neGA mumupunga41, zvigumisiro zvedu zvinoratidza kuti kuchinja kwepamusoro-soro kweArabidopsis EL1 homologue (AEL1-4) hakuderedza RGA phosphorylation. Mukubvumirana nemigumisiro yedu, ichangoburwa phosphoproteomic kudzidza kushandisa Arabidopsis AEL overexpressing mitsara uye ael katatu mutant haina kuratidza chero mapuroteni eDELLA se substrates yeaya kinases56. Patakagadzirira chinyorwa, zvakashumwa kuti GSK3, gene encoding GSK3 / SHAGGY-like kinase mugorosi (Triticum aestivum), inogona phosphorylate DELLA (Rht-B1b)57, kunyange zvazvo phosphorylation yeRht-B1b neGSK3 haina kusimbiswa muplanta. In vitro enzymatic reactions muhupo hweGSK3 inoteverwa nehukuru spectrometry kuongorora yakaratidza nzvimbo nhatu dzephosphorylation dziri pakati peDELLA neGRAS domains yeRht-B1b (Supplementary Fig. 3). Serine kune alanine inotsiva pane ese matatu phosphorylation saiti yakakonzera kuderera kweRht-B1b chiitiko mune transgenic gorosi, zvinoenderana nezvatakawana kuti alanine inotsiva muPep2 RGA yakaderedza RGA basa. Nekudaro, in vitro protein degradation assays yakawedzera kuratidza kuti phosphorylation inogona zvakare kudzikamisa Rht-B1b57. Izvi zvinopesana nemhedzisiro yedu inoratidza kuti alanine inotsiva muPep2 RGA haishandure kugadzikana kwayo muplanta. GSK3 mugorosi inonzi ortholog ye brassinosteroid-insensitive protein 2 (BIN2) muArabidopsis 57. BIN2 inodzora chakaipa chechiratidzo cheBR, uye BR inoshandisa nzira yayo yekuratidzira kuburikidza nekukonzera BIN2 kuparara 58. Takaratidza kuti BR kurapa haideredzi RGA kugadzikana mazinga 59 kana phosphory 2 phosphory (phosphory). zvichiratidza kuti RGA haifanire kuve phosphorylated neBIN2.
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