Mālama ʻia nā protein DELLAnā mea hoʻoponopono uluka mea nui i ka ulu ʻana o nā mea kanu i ka pane ʻana i nā hōʻailona o loko a me waho. Ma ke ʻano he mea hoʻoponopono transcriptional, hoʻopaʻa nā DELLA i nā mea transcription (TFs) a me ka histone H2A ma o kā lākou mau kāʻei GRAS a ua kiʻi ʻia e hana i nā mea hoʻolaha. Ua hōʻike ʻia nā haʻawina hou e hoʻoponopono ʻia ka paʻa o DELLA ma hope o ka unuhi ʻana e nā ʻano hana ʻelua: polyubiquitination i hoʻokomo ʻia e ka hormone mea kanu.giberellin, ka mea e alakaʻi i ko lākou degradation wikiwiki, a conjugation me ka liʻiliʻi ubiquitin-like modifier (SUMO), e hoʻonui i kā lākou hōʻiliʻili. Eia kekahi, ua hoʻoponopono ikaika ʻia ka hana DELLA e ʻelua mau hana glycosylation kūʻokoʻa: Hoʻonui ka O-fucosylation i ka pilina DELLA-TF, ʻoiai ʻo O-linked N-acetylglucosamine (O-GlcNAc) hoʻololi e kāohi i ka pilina DELLA-TF. Eia naʻe, ʻaʻole maopopo ka hana o ka DELLA phosphorylation e like me nā haʻawina mua i hōʻike i nā hopena kū'ē, me ka manaʻo o kekahi e hāpai a hoʻopaʻa paha ka phosphorylation i ka degradation DELLA a me nā mea ʻē aʻe e ʻōlelo ana ʻaʻole pili ka phosphorylation i ko lākou kūpaʻa. Maʻaneʻi,ʻike mākou i nā pūnaewele phosphorylation i ka GA1-3 repressor (RGA), AtDELLA, i hoʻomaʻemaʻeʻia mai ka Arabidopsis thaliana e ka spectrometry nui a hōʻike i ka phosphorylation oʻelua RGA peptides i nā'āpana PolyS a me PolyS / T e hoʻonui i ka hana RGA ma ka hoʻolahaʻana i ka H2A paʻa a me ka hui RGA me nā mea hoʻolaha. ʻO ka mea nui, ʻaʻole i pili ka phosphorylation i ka pilina RGA-TF a i ʻole ka paʻa ʻana o RGA. Hōʻike kā mākou noiʻi ʻana i kahi ʻano molekala e hoʻoulu ai ka phosphorylation i ka hana DELLA.
Ua hōʻike ʻia kā mākou loiloi spectrometric mass ʻo Pep1 a me Pep2 i phosphorylated nui loa i RGA ma ke kāʻei ʻo GA-deficient Ga1. Ma waho aʻe o kēia haʻawina, ua hōʻike pū nā haʻawina phosphoproteomic i ka phosphorylation Pep1 ma RGA, ʻoiai ʻaʻole i aʻo ʻia kāna kuleana53,54,55. I ka hoʻohālikelike ʻana, ʻaʻole i wehewehe mua ʻia ka phosphorylation Pep2 no ka mea hiki ke ʻike ʻia kēia peptide me ka hoʻohana ʻana i ka transgene RGAGKG. ʻOiai ʻo ka mutation m1A, ka mea i hoʻopau i ka phosphorylation Pep1, hoʻemi iki wale i ka hana RGA i ka mea kanu, he hopena hoʻohui ia i ka wā i hui pū ʻia me ka m2A i ka hōʻemi ʻana i ka hana RGA (Supplementary Figure 6). ʻO ka mea nui, ua hoʻemi nui ʻia ka phosphorylation Pep1 i ka mutant sly1 i hoʻonui ʻia e GA i hoʻohālikelike ʻia me ga1, e hōʻike ana e hoʻolaha ana ʻo GA i ka dephosphorylation RGA, e hōʻemi ana i kāna hana. ʻO ka mīkini e hoʻopau ai ʻo GA i ka phosphorylation RGA e pono ai ka hoʻokolokolo hou. ʻO kekahi mea hiki ke loaʻa kēia ma o ka hoʻoponopono ʻana i kahi kinase protein i ʻike ʻole ʻia. ʻOiai ua hōʻike ʻia nā haʻawina e hoʻohaʻahaʻa ʻia ka ʻōlelo o ka CK1 protein kinase EL1 e GA i ka laiki41, hōʻike kā mākou mau hopena i ka hoʻololi ʻana o ka homologue Arabidopsis EL1 (AEL1-4) ʻaʻole e hōʻemi i ka phosphorylation RGA. I ka ʻae ʻana me kā mākou hopena, kahi noiʻi phosphoproteomic hou e hoʻohana ana i nā laina overexpressing Arabidopsis AEL a me kahi ael triple mutant ʻaʻole i ʻike i nā protein DELLA ma ke ʻano he substrate o kēia mau kinases56. I ko mākou hoʻomākaukau ʻana i ka manuscript, ua hōʻike ʻia ʻo GSK3, ka gene e hoʻopili ana i kahi kinase GSK3/SHAGGY-like i ka palaoa (Triticum aestivum), hiki ke phosphorylate DELLA (Rht-B1b) 57, ʻoiai ʻaʻole i hōʻoia ʻia ka phosphorylation o Rht-B1b e GSK3 i loko o ka mea kanu. In vitro enzymatic reactions i ke alo o GSK3 ukali e ka nuipa spectrometry kālailai hōʻike 'ekolu phosphorylation paena i loaʻa ma waena o ka DELLA a me GRAS domain o Rht-B1b (Supplementary Fig. 3). ʻO Serine i ka alanine substitutions ma nā wahi phosphorylation ʻekolu i hoʻemi ʻia ka hana Rht-B1b i ka palaoa transgenic, e like me kā mākou ʻike ʻana i ka hoʻololi ʻana o ka alanine ma Pep2 RGA i ka hana RGA. Eia nō naʻe, ua hōʻike hou ʻia nā hōʻike hoʻohaʻahaʻa in vitro protein hiki i ka phosphorylation ke hoʻopaʻa i ka Rht-B1b57. He ʻokoʻa kēia i kā mākou hopena e hōʻike ana ʻaʻole i hoʻololi ke pani alanine i ka Pep2 RGA i kona kūpaʻa i ka mea kanu. ʻO GSK3 i ka palaoa he ortholog o ka brassinosteroid-insensitive protein 2 (BIN2) ma Arabidopsis 57. ʻO BIN2 kahi mea hoʻoponopono maikaʻi ʻole o ka hōʻailona BR, a hoʻāla ʻo BR i kona ala hōʻailona ma o ka hoʻohaʻahaʻa ʻana i ka BIN2 58. phosphorylated e BIN2.
Ua kālailai ʻia nā ʻikepili quantitative a pau me ka Excel, a ua hoʻoholo ʻia nā ʻokoʻa koʻikoʻi me ka hoʻohana ʻana i ka t-test a Student. ʻAʻole i hoʻohana ʻia nā ʻano helu helu no ka hoʻoholo mua ʻana i ka nui o ka laʻana. ʻAʻohe ʻikepili i hoʻokaʻawale ʻia mai ka nānā ʻana; ʻaʻole i hoʻohālikelike ʻia ka hoʻokolohua; a ua ʻike ka poʻe noiʻi i ka hoʻokaʻawale ʻana i ka wā o ka hoʻokolohua a me ka loiloi hopena. Hāʻawi ʻia nā laʻana nui ma nā kaʻao kiʻi a me nā faila ʻike maka.
Ka manawa hoʻouna: Apr-15-2025