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- EMDB-11789: RUVBL1-RUVBL2 heterohexameric ring after binding of RNA helicase DHX34 -
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基本情報
登録情報 | データベース: EMDB / ID: EMD-11789 | ||||||||||||
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タイトル | RUVBL1-RUVBL2 heterohexameric ring after binding of RNA helicase DHX34 | ||||||||||||
![]() | Cryo-EM structure of RUVBL1-RUVBL2 ATPase core | ||||||||||||
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![]() | RUVBL1-RUVBL2 / DHX34 / Nonsense-Mediated mRNA Decay / RNA degradation / Cryo-EM / TRANSLATION | ||||||||||||
機能・相同性 | ![]() promoter-enhancer loop anchoring activity / telomerase RNA localization to Cajal body / regulation of DNA strand elongation / positive regulation of telomere maintenance in response to DNA damage / establishment of protein localization to chromatin / R2TP complex / dynein axonemal particle / Swr1 complex / RPAP3/R2TP/prefoldin-like complex / Ino80 complex ...promoter-enhancer loop anchoring activity / telomerase RNA localization to Cajal body / regulation of DNA strand elongation / positive regulation of telomere maintenance in response to DNA damage / establishment of protein localization to chromatin / R2TP complex / dynein axonemal particle / Swr1 complex / RPAP3/R2TP/prefoldin-like complex / Ino80 complex / regulation of double-strand break repair / box C/D snoRNP assembly / protein folding chaperone complex / NuA4 histone acetyltransferase complex / regulation of chromosome organization / TFIID-class transcription factor complex binding / regulation of DNA replication / MLL1 complex / regulation of embryonic development / Telomere Extension By Telomerase / positive regulation of double-strand break repair via homologous recombination / RNA polymerase II core promoter sequence-specific DNA binding / regulation of DNA repair / Deposition of new CENPA-containing nucleosomes at the centromere / telomere maintenance / DNA helicase activity / positive regulation of DNA repair / TBP-class protein binding / DNA Damage Recognition in GG-NER / cellular response to estradiol stimulus / chromatin DNA binding / Formation of the beta-catenin:TCF transactivating complex / euchromatin / negative regulation of canonical Wnt signaling pathway / ADP binding / beta-catenin binding / nuclear matrix / UCH proteinases / cellular response to UV / transcription corepressor activity / unfolded protein binding / positive regulation of canonical Wnt signaling pathway / nucleosome / protein folding / HATs acetylate histones / ATPase binding / spermatogenesis / regulation of apoptotic process / DNA recombination / DNA helicase / transcription coactivator activity / Ub-specific processing proteases / regulation of cell cycle / protein stabilization / nuclear speck / ciliary basal body / chromatin remodeling / cadherin binding / RNA polymerase II cis-regulatory region sequence-specific DNA binding / ribonucleoprotein complex / cell division / DNA repair / centrosome / regulation of transcription by RNA polymerase II / regulation of DNA-templated transcription / positive regulation of DNA-templated transcription / protein homodimerization activity / positive regulation of transcription by RNA polymerase II / ATP hydrolysis activity / extracellular exosome / nucleoplasm / ATP binding / identical protein binding / nucleus / membrane / cytosol / cytoplasm 類似検索 - 分子機能 | ||||||||||||
生物種 | ![]() | ||||||||||||
手法 | 単粒子再構成法 / クライオ電子顕微鏡法 / 解像度: 4.18 Å | ||||||||||||
![]() | Lopez-Perrote A / Rodriguez CF / Llorca O | ||||||||||||
資金援助 | ![]()
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![]() | ![]() タイトル: Regulation of RUVBL1-RUVBL2 AAA-ATPases by the nonsense-mediated mRNA decay factor DHX34, as evidenced by Cryo-EM. 著者: Andres López-Perrote / Nele Hug / Ana González-Corpas / Carlos F Rodríguez / Marina Serna / Carmen García-Martín / Jasminka Boskovic / Rafael Fernandez-Leiro / Javier F Caceres / Oscar Llorca / ![]() ![]() 要旨: Nonsense-mediated mRNA decay (NMD) is a surveillance pathway that degrades aberrant mRNAs and also regulates the expression of a wide range of physiological transcripts. RUVBL1 and RUVBL2 AAA-ATPases ...Nonsense-mediated mRNA decay (NMD) is a surveillance pathway that degrades aberrant mRNAs and also regulates the expression of a wide range of physiological transcripts. RUVBL1 and RUVBL2 AAA-ATPases form an hetero-hexameric ring that is part of several macromolecular complexes such as INO80, SWR1, and R2TP. Interestingly, RUVBL1-RUVBL2 ATPase activity is required for NMD activation by an unknown mechanism. Here, we show that DHX34, an RNA helicase regulating NMD initiation, directly interacts with RUVBL1-RUVBL2 in vitro and in cells. Cryo-EM reveals that DHX34 induces extensive changes in the N-termini of every RUVBL2 subunit in the complex, stabilizing a conformation that does not bind nucleotide and thereby down-regulates ATP hydrolysis of the complex. Using ATPase-deficient mutants, we find that DHX34 acts exclusively on the RUVBL2 subunits. We propose a model, where DHX34 acts to couple RUVBL1-RUVBL2 ATPase activity to the assembly of factors required to initiate the NMD response. | ||||||||||||
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注釈 | Cryo-EM structure of RUVBL1-RUVBL2 ATPase core | ||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
投影像・断面図 | 画像のコントロール
画像は Spider により作成 | ||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
ボクセルのサイズ | X=Y=Z: 1.047 Å | ||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
密度 |
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対称性 | 空間群: 1 | ||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
詳細 | EMDB XML:
CCP4マップ ヘッダ情報:
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試料の構成要素
-全体 : RUVBL1-RUVBL2 heterohexameric ring after binding of RNA helicase DHX34
全体 | 名称: RUVBL1-RUVBL2 heterohexameric ring after binding of RNA helicase DHX34 |
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要素 |
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-超分子 #1: RUVBL1-RUVBL2 heterohexameric ring after binding of RNA helicase DHX34
超分子 | 名称: RUVBL1-RUVBL2 heterohexameric ring after binding of RNA helicase DHX34 タイプ: complex / ID: 1 / 親要素: 0 / 含まれる分子: #1-#2 詳細: Structure of the RUVBL1-RUVBL2 hetero-hexameric ring region after the interaction of RNA helicase DHX34 |
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由来(天然) | 生物種: ![]() |
分子量 | 理論値: 0.44928 kDa/nm |
-分子 #1: RuvB-like 1
分子 | 名称: RuvB-like 1 / タイプ: protein_or_peptide / ID: 1 / コピー数: 3 / 光学異性体: LEVO / EC番号: DNA helicase |
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由来(天然) | 生物種: ![]() |
分子量 | 理論値: 52.710406 KDa |
組換発現 | 生物種: ![]() ![]() |
配列 | 文字列: HHHHHHHHSS GENLYFQGSH MKIEEVKSTT KTQRIASHSH VKGLGLDESG LAKQAASGLV GQENAREACG VIVELIKSKK MAGRAVLLA GPPGTGKTAL ALAIAQELGS KVPFCPMVGS EVYSTEIKKT EVLMENFRRA IGLRIKETKE VYEGEVTELT P CETENPMG ...文字列: HHHHHHHHSS GENLYFQGSH MKIEEVKSTT KTQRIASHSH VKGLGLDESG LAKQAASGLV GQENAREACG VIVELIKSKK MAGRAVLLA GPPGTGKTAL ALAIAQELGS KVPFCPMVGS EVYSTEIKKT EVLMENFRRA IGLRIKETKE VYEGEVTELT P CETENPMG GYGKTISHVI IGLKTAKGTK QLKLDPSIFE SLQKERVEAG DVIYIEANSG AVKRQGRCDT YATEFDLEAE EY VPLPKGD VHKKKEIIQD VTLHDLDVAN ARPQGGQDIL SMMGQLMKPK KTEITDKLRG EINKVVNKYI DQGIAELVPG VLF VDEVHM LDIECFTYLH RALESSIAPI VIFASNRGNC VIRGTEDITS PHGIPLDLLD RVMIIRTMLY TPQEMKQIIK IRAQ TEGIN ISEEALNHLG EIGTKTTLRY SVQLLTPANL LAKINGKDSI EKEHVEEISE LFYDAKSSAK ILADQQDKYM K UniProtKB: RuvB-like 1 |
-分子 #2: RuvB-like 2
分子 | 名称: RuvB-like 2 / タイプ: protein_or_peptide / ID: 2 / コピー数: 3 / 光学異性体: LEVO / EC番号: DNA helicase |
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由来(天然) | 生物種: ![]() |
分子量 | 理論値: 53.047488 KDa |
組換発現 | 生物種: ![]() ![]() |
配列 | 文字列: MADLNWISAG HAIADVGTMA TVTATTKVPE IRDVTRIERI GAHSHIRGLG LDDALEPRQA SQGMVGQLAA RRAAGVVLEM IREGKIAGR AVLIAGQPGT GKTAIAMGMA QALGPDTPFT AIAGSEIFSL EMSKTEALTQ AFRRSIGVRI KEETEIIEGE V VEIQIDRP ...文字列: MADLNWISAG HAIADVGTMA TVTATTKVPE IRDVTRIERI GAHSHIRGLG LDDALEPRQA SQGMVGQLAA RRAAGVVLEM IREGKIAGR AVLIAGQPGT GKTAIAMGMA QALGPDTPFT AIAGSEIFSL EMSKTEALTQ AFRRSIGVRI KEETEIIEGE V VEIQIDRP ATGTGSKVGK LTLKTTEMET IYDLGTKMIE SLTKDKVQAG DVITIDKATG KISKLGRSFT RARDYDAMGS QT KFVQCPD GELQKRKEVV HTVSLHEIDV INSRTQGFLA LFSGDTGEIK SEVREQINAK VAEWREEGKA EIIPGVLFID EVH MLDIES FSFLNRALES DMAPVLIMAT NRGITRIRGT SYQSPHGIPI DLLDRLLIVS TTPYSEKDTK QILRIRCEEE DVEM SEDAY TVLTRIGLET SLRYAIQLIT AASLVCRKRK GTEVQVDDIK RVYSLFLDES RSTQYMKEYQ DAFLFNELKG ETMDT S UniProtKB: RuvB-like 2 |
-分子 #3: ADENOSINE-5'-DIPHOSPHATE
分子 | 名称: ADENOSINE-5'-DIPHOSPHATE / タイプ: ligand / ID: 3 / コピー数: 3 / 式: ADP |
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分子量 | 理論値: 427.201 Da |
Chemical component information | ![]() ChemComp-ADP: |
-実験情報
-構造解析
手法 | クライオ電子顕微鏡法 |
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![]() | 単粒子再構成法 |
試料の集合状態 | particle |
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試料調製
濃度 | 0.35 mg/mL |
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緩衝液 | pH: 7.4 / 構成要素 - 式: Tris-NaCl / 構成要素 - 名称: TBS / 詳細: 50 mM Tris-HCl pH 7.4, 150 mM NaCl |
凍結 | 凍結剤: ETHANE / チャンバー内湿度: 95 % / チャンバー内温度: 277.15 K / 装置: FEI VITROBOT MARK IV |
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電子顕微鏡法
顕微鏡 | FEI TITAN KRIOS |
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撮影 | フィルム・検出器のモデル: GATAN K2 SUMMIT (4k x 4k) 検出モード: COUNTING / 撮影したグリッド数: 1 / 実像数: 3047 / 平均電子線量: 48.1 e/Å2 |
電子線 | 加速電圧: 300 kV / 電子線源: ![]() |
電子光学系 | 倍率(補正後): 47756 / 照射モード: FLOOD BEAM / 撮影モード: BRIGHT FIELD / Cs: 2.7 mm / 最大 デフォーカス(公称値): 0.003 µm / 最小 デフォーカス(公称値): 0.0015 µm |
試料ステージ | 試料ホルダーモデル: FEI TITAN KRIOS AUTOGRID HOLDER ホルダー冷却材: NITROGEN |
実験機器 | ![]() モデル: Titan Krios / 画像提供: FEI Company |