Phosphorylation imayendetsa master kukula kwa DELLA mu Arabidopsis polimbikitsa kuyanjana kwa histone H2A ndi chromatin.

Mapuloteni a DELLA amasungidwa bwinozowongolera kukulazomwe zimagwira ntchito yayikulu pakuwongolera kukula kwa mbewu potengera zomwe zikuchitika mkati ndi chilengedwe. DELLA imagwira ntchito ngati chowongolera cholembera ndipo amalembedwa kuti azitsatira otsatsa pomanga zinthu zolembera (TFs) ndi histone H2A kudzera mu domain yake ya GRAS. Kafukufuku waposachedwapa wasonyeza kuti kukhazikika kwa DELLA kumayendetsedwa pambuyo pomasulira pogwiritsa ntchito njira ziwiri: polyubiquitination yoyambitsidwa ndi phytohormone gibberellin, yomwe imayambitsa kuwonongeka kwake mofulumira, ndi kugwirizanitsa kwa osintha ang'onoang'ono a ubiquitin-like modifiers (SUMO) kuti awonjezere kudzikundikira kwake. Kuphatikiza apo, ntchito ya DELLA imayendetsedwa mwamphamvu ndi ma glycosylations awiri osiyana: kuyanjana kwa DELLA-TF kumakulitsidwa ndi O-fucosylation koma kuletsedwa ndi kusinthidwa kwa O-linked N-acetylglucosamine (O-GlcNAc). Komabe, ntchito ya DELLA phosphorylation imakhalabe yosadziwika bwino, monga momwe kafukufuku wam'mbuyomu adawonetsa zotsatira zotsutsana, kuyambira zomwe zikuwonetsa kuti phosphorylation imalimbikitsa kapena kuchepetsa kuwonongeka kwa DELLA kwa ena kusonyeza kuti phosphorylation sichikhudza kukhazikika kwake. Apa, timazindikira malo a phosphorylation mu REPRESSORgawo 1-3(RGA, AtDELLA) yoyeretsedwa kuchokera ku Arabidopsis thaliana ndi kusanthula kwakukulu kwa ma spectrometry ndikuwonetsa kuti phosphorylation ya ma peptides awiri a RGA kumadera a PolyS ndi PolyS/T amalimbikitsa H2A kumanga ndi kupititsa patsogolo ntchito ya RGA. Mgwirizano wa RGA ndi omwe amalimbikitsa. Makamaka, phosphorylation sichikhudza kuyanjana kwa RGA-TF kapena kukhazikika kwa RGA. Kafukufuku wathu akuwonetsa momwe ma cell a phosphorylation amathandizira ntchito ya DELLA.
Kuti timvetsetse ntchito ya phosphorylation pakuwongolera ntchito ya DELLA, ndikofunikira kuzindikira malo a DELLA phosphorylation mu vivo ndikuwunika magwiridwe antchito muzomera. Mwa kuyeretsedwa kogwirizana kwa zotsalira za mbewu zotsatiridwa ndi kusanthula kwa MS/MS, tidazindikira ma phosphosite angapo mu RGA. Pansi pa kuchepa kwa GA, RHA phosphorylation imawonjezeka, koma phosphorylation sichimakhudza kukhazikika kwake. Chofunika kwambiri, kuyesa kwa Co-IP ndi ChIP-qPCR kunawonetsa kuti phosphorylation ku PolyS/T dera la RGA imalimbikitsa kuyanjana kwake ndi H2A komanso kuyanjana ndi olimbikitsa omwe akufuna, kuwulula njira yomwe phosphorylation imapangitsa RGA kugwira ntchito.
RGA imalembedwanso kuti igwirizane ndi chromatin kudzera mumgwirizano wa subdomain ya LHR1 ndi TF kenako imamanga ku H2A kudzera m'chigawo chake cha PolyS/T ndi PFYRE subdomain, kupanga H2A-RGA-TF complex kuti akhazikitse RGA. Phosphorylation ya Pep 2 m'chigawo cha PolyS/T pakati pa dera la DELLA ndi dera la GRAS ndi kinase yosadziwika imapangitsa RGA-H2A kumanga. Puloteni yosinthika ya rgam2A imathetsa phosphorylation ya RGA ndipo imatenga mapuloteni osiyanasiyana kuti asokoneze kumanga kwa H2A. Izi zimabweretsa kusokonezeka kwa kuyanjana kwakanthawi kwa TF-rgam2A ndikusiyanitsidwa kwa rgam2A kuchokera ku chandamale cha chromatin. Chiwerengerochi chikungowonetsa kuponderezedwa ndi RGA-mediated transcriptal. Chitsanzo chofananacho chikhoza kufotokozedwa pa RGA-mediated transcriptional activation, kupatula kuti zovuta za H2A-RGA-TF zingalimbikitse kulembedwa kwa jini ndi dephosphorylation ya rgam2A ingachepetse kulembedwa. Chithunzi chosinthidwa kuchokera ku Huang et al.21.
Deta yonse yochulukira idawunikidwa pogwiritsa ntchito Excel, ndipo kusiyana kwakukulu kunatsimikiziridwa pogwiritsa ntchito mayeso a Student t. Palibe njira zowerengera zomwe zidagwiritsidwa ntchito kuti zitsimikizire kukula kwachitsanzo. Palibe deta yomwe idachotsedwa pakuwunika; kuyesera sikunali kwachisawawa; ofufuzawo sanali akhungu pakugawa deta panthawi yoyesera ndikuwunika zotsatira. Kukula kwachitsanzo kukuwonetsedwa mu nthano yachifanizo ndi fayilo yochokera.
Kuti mumve zambiri za kapangidwe ka kafukufukuyu, onani Natural Portfolio Report Abstract yokhudzana ndi nkhaniyi.
Deta ya Mass spectrometry proteomics yathandizidwa ku ProteomeXchange consortium kudzera munkhokwe ya anzawo a PRIDE66 yokhala ndi chizindikiritso cha dataset PXD046004. Zina zonse zomwe zapezedwa panthawi ya phunziroli zimaperekedwa mu Supplementary Information, Supplementary Data Files, ndi Raw Data Files. Zomwe zachokera m'nkhaniyi zaperekedwa.