Phosphorylation inomutsa chinodzora kukura kwemasero cheDELLA, zvichiita kuti histone H2A ibatane nechromatin muArabidopsis.

Mapuroteni eDella anochengetwavanodzora kukuraizvo zvinoita basa guru mukukura kwezvirimwa zvichienderana nezviratidzo zvemukati nekunze. Sevanodzora transcriptional, DELLAs vanosunga kune transcription factors (TFs) uye histone H2A kuburikidza neGRAS domains dzavo uye vanotorwa kuti vashande pane vanokurudzira. Ongororo dzichangoburwa dzakaratidza kuti DELLA stability inodzorwa mushure mekushandurwa nenzira mbiri: polyubiquitination inokonzerwa ne plant hormone.gibberellin, izvo zvinotungamira mukuora kwavo nekukurumidza, uye kubatana ne small ubiquitin-like modifier (SUMO), iyo inowedzera kuunganidzwa kwavo. Uyezve, basa reDELLA rinodzorwa nenzira mbiri dzakasiyana dzeglycosylation: O-fucosylation inowedzera kudyidzana kweDELLA-TF, nepo O-linked N-acetylglucosamine (O-GlcNAc) kuchidzivisa kudyidzana kweDELLA-TF. Zvisinei, basa reDELLA phosphorylation harina kujeka sezvo zvidzidzo zvekare zvakaratidza mhedzisiro inopikisana, zvimwe zvichiratidza kuti phosphorylation inokurudzira kana kudzvinyirira kuparadzwa kweDELLA uye zvimwe zvichiratidza kuti phosphorylation haikanganisi kugadzikana kwavo. Pano, tinoona nzvimbo dzephosphorylation muGA1-3 repressor (RGA), AtDELLA, yakacheneswa kubva kuArabidopsis thaliana ne mass spectrometry uye inoratidza kuti phosphorylation yemapeptide maviri eRGA munzvimbo dzePolyS nePolyS/T inowedzera kushandira kweRGA nekusimudzira kubatana kweH2A uye kubatana kweRGA nevanokurudzira chinangwa. Zvinoshamisa kuti, phosphorylation haina kukanganisa kudyidzana kweRGA-TF kana kugadzikana kweRGA. Chidzidzo chedu chinoratidza nzira yemamorekuru iyo phosphorylation inokonzera nayo basa reDELLA.
Ongororo yedu yekuongorora mass spectrometric yakaratidza kuti Pep1 nePep2 zvaive nephosphorylation yakawanda muRGA muGa1 isina GA. Pamusoro pechidzidzo ichi, zvidzidzo zvephosphoproteomic zvakaratidzawo phosphorylation yePep1 muRGA, kunyangwe basa rayo risati radzidzwa53,54,55. Kusiyana neizvi, phosphorylation yePep2 haisati yatsanangurwa kare sezvo iyi peptide yaigona kuonekwa chete uchishandisa RGAGKG transgene. Kunyangwe m1A mutation, iyo yakabvisa phosphorylation yePep1, yakangoderedza zvishoma basa reRGA muplanta, yaive nemhedzisiro yekuwedzera kana yakabatanidzwa nem2A mukuderedza basa reRGA (Mufananidzo Wekuwedzera 6). Zvakakosha, phosphorylation yePep1 yakaderedzwa zvakanyanya muGA-enhanced sly1 mutant zvichienzaniswa nega1, zvichiratidza kuti GA inokurudzira RGA dephosphorylation, ichideredza basa rayo. Nzira iyo GA inodzvinyirira RGA phosphorylation inoda kuferefetwa kwakawanda. Imwe mukana ndeyekuti izvi zvinoitwa kuburikidza nekudzora protein kinase isingazivikanwe. Kunyangwe zvidzidzo zvakaratidza kuti kuburitswa kweCK1 protein kinase EL1 kunoderedzwa neGA mu rice41, zvatakawana zvinoratidza kuti kuchinja kweArabidopsis EL1 homologue (AEL1-4) kwakanyanya hakuderedzi RGA phosphorylation. Zvichienderana nezvatakawana, chidzidzo chechangobva kuitwa chephosphoproteomic chichishandisa Arabidopsis AEL overexpressing lines uye ael triple mutant hachina kuratidza chero mapuroteni eDELLA sezvikamu zvema kinases aya56. Patakagadzirira chinyorwa, zvakanzi GSK3, gene rinonyora GSK3/SHAGGY-like kinase mugorosi (Triticum aestivum), rinogona phosphorylate DELLA (Rht-B1b)57, kunyangwe phosphorylation yeRht-B1b neGSK3 isina kusimbiswa muplanta. Maitiro e enzyme mu vitro pamberi peGSK3 zvichiteverwa nekuongorora kwe mass spectrometry kwakaratidza nzvimbo nhatu dze phosphorylation dziri pakati peDELLA neGRAS domains dzeRht-B1b (Supplementary Fig. 3). Kutsiva kwe serine kuenda ku alanine panzvimbo dzese nhatu dze phosphorylation kwakaita kuti basa reRht-B1b riderere mu transgenic wheat, zvichienderana nezvatakawana zvekuti kutsiva alanine muPep2 RGA kwakaderedza basa reRGA. Zvisinei, kuongororwa kwekudzikira kwemapuroteni mu vitro kwakaratidzawo kuti phosphorylation inogonawo kudzikamisa Rht-B1b57. Izvi zvakasiyana nezvatakawana zvinoratidza kuti kutsiva alanine muPep2 RGA hakuchinji kugadzikana kwayo mu planta. GSK3 mugorosi ipuroteni isinganzwe brassinosteroid 2 (BIN2) muArabidopsis 57. BIN2 inodzora zviratidzo zveBR, uye BR inomutsa nzira yayo yezviratidzo nekukonzera kuora kweBIN2 58. Takaratidza kuti kurapwa kweBR hakuderedzi kugadzikana kweRGA 59 kana mazinga ephosphorylation muArabidopsis (Mufananidzo Wekuwedzera 2), zvichiratidza kuti RGA haigone kufosforasiwa neBIN2.
Data rese rehuwandu rakaongororwa neExcel, uye mutsauko wakakosha wakaongororwa uchishandisa bvunzo ye t yemudzidzi. Hapana nzira dzehuwandu dzakashandiswa kuona saizi yesampuro pachine nguva. Hapana data rakabviswa muongororo; kuyedza kwacho hakuna kurongeka; uye vaongorori vaiziva nezvekugoverwa kwesampuro panguva yekuyedza uye kuongororwa kwemhedzisiro. Sampuro dzesampuro dzinopihwa mungano dzemifananidzo uye mumafaira edata asina kurongeka.