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基本情報
登録情報 | データベース: EMDB / ID: EMD-2664 | |||||||||
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タイトル | Structure of SMG1C-UPF1 complex, comprising SMG1 kinase, SMG8, SMG9 and UPF1 | |||||||||
![]() | Reconstruction of the SMG1C-UPF1 complex, comprising SMG1, SMG8, SMG9 and UPF1 | |||||||||
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![]() | NMD / SMG1 / SMG8 / SMG9 / UPF1 / PIKK / RNA degradation | |||||||||
機能・相同性 | ![]() positive regulation of mRNA cis splicing, via spliceosome / supraspliceosomal complex / RNA metabolic process / exon-exon junction complex / regulation of protein kinase activity / telomere maintenance via semi-conservative replication / positive regulation of mRNA catabolic process / cell cycle phase transition / diacylglycerol-dependent serine/threonine kinase activity / regulation of translational termination ...positive regulation of mRNA cis splicing, via spliceosome / supraspliceosomal complex / RNA metabolic process / exon-exon junction complex / regulation of protein kinase activity / telomere maintenance via semi-conservative replication / positive regulation of mRNA catabolic process / cell cycle phase transition / diacylglycerol-dependent serine/threonine kinase activity / regulation of translational termination / histone mRNA catabolic process / eye development / chromatoid body / 3'-UTR-mediated mRNA destabilization / regulation of telomere maintenance / nuclear-transcribed mRNA catabolic process, nonsense-mediated decay / telomeric DNA binding / phosphatidylinositol phosphate biosynthetic process / nuclear-transcribed mRNA catabolic process / cellular response to interleukin-1 / Nonsense Mediated Decay (NMD) independent of the Exon Junction Complex (EJC) / mRNA export from nucleus / Nonsense Mediated Decay (NMD) enhanced by the Exon Junction Complex (EJC) / helicase activity / P-body / brain development / peptidyl-serine phosphorylation / heart development / cellular response to lipopolysaccharide / protein autophosphorylation / DNA helicase / in utero embryonic development / double-stranded DNA helicase activity / eukaryotic translation initiation factor 2alpha kinase activity / chromosome, telomeric region / 3-phosphoinositide-dependent protein kinase activity / DNA-dependent protein kinase activity / ribosomal protein S6 kinase activity / histone H3S10 kinase activity / histone H2AXS139 kinase activity / histone H3S28 kinase activity / histone H4S1 kinase activity / histone H2BS14 kinase activity / histone H3T3 kinase activity / histone H2AS121 kinase activity / Rho-dependent protein serine/threonine kinase activity / histone H2BS36 kinase activity / histone H3S57 kinase activity / histone H2AT120 kinase activity / AMP-activated protein kinase activity / histone H2AS1 kinase activity / histone H3T6 kinase activity / histone H3T11 kinase activity / histone H3T45 kinase activity / DNA replication / non-specific serine/threonine protein kinase / protein kinase activity / RNA helicase activity / RNA helicase / protein serine kinase activity / DNA repair / protein serine/threonine kinase activity / DNA damage response / chromatin binding / protein-containing complex binding / negative regulation of apoptotic process / chromatin / perinuclear region of cytoplasm / ATP hydrolysis activity / RNA binding / zinc ion binding / nucleoplasm / ATP binding / metal ion binding / identical protein binding / nucleus / cytosol / cytoplasm 類似検索 - 分子機能 | |||||||||
生物種 | ![]() | |||||||||
手法 | 単粒子再構成法 / ネガティブ染色法 / 解像度: 20.9 Å | |||||||||
![]() | Melero R / Uchiyama A / Castano R / Kataoka N / Kurosawa H / Ohno S / Yamashita A / Llorca O | |||||||||
![]() | ![]() タイトル: Structures of SMG1-UPFs complexes: SMG1 contributes to regulate UPF2-dependent activation of UPF1 in NMD. 著者: Roberto Melero / Akiko Uchiyama / Raquel Castaño / Naoyuki Kataoka / Hitomi Kurosawa / Shigeo Ohno / Akio Yamashita / Oscar Llorca / ![]() ![]() 要旨: SMG1, a PI3K-related kinase, plays a critical role in nonsense-mediated mRNA decay (NMD) in mammals. SMG1-mediated phosphorylation of the UPF1 helicase is an essential step during NMD initiation. ...SMG1, a PI3K-related kinase, plays a critical role in nonsense-mediated mRNA decay (NMD) in mammals. SMG1-mediated phosphorylation of the UPF1 helicase is an essential step during NMD initiation. Both SMG1 and UPF1 are presumably activated by UPF2, but this regulation is incompletely understood. Here we reveal that SMG1C (a complex containing SMG1, SMG8, and SMG9) contributes to regulate NMD by recruiting UPF1 and UPF2 to distinct sites in the vicinity of the kinase domain. UPF2 binds SMG1 in an UPF1-independent manner in vivo, and the SMG1C-UPF2 structure shows UPF2 recognizes the FRB domain, a region that regulates the related mTOR kinase. The molecular architectures of several SMG1C-UPFs complexes, obtained by combining electron microscopy with in vivo and in vitro interaction analyses, competition experiments, and mutations, suggest that UPF2 can be transferred to UPF1 within SMG1C, inducing UPF2-dependent conformational changes required to activate UPF1 within an SMG1C-UPF1-UPF2 complex. | |||||||||
履歴 |
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構造の表示
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構造ビューア | EMマップ: ![]() ![]() ![]() |
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ヘッダ (付随情報) | ![]() ![]() | 14.1 KB 14.1 KB | 表示 表示 | ![]() |
画像 | ![]() | 53.4 KB | ||
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注釈 | Reconstruction of the SMG1C-UPF1 complex, comprising SMG1, SMG8, SMG9 and UPF1 | ||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
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: 2.84 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 : SMG1C-UPF1 complex, comprising SMG1 kinase, SMG8, SMG9 and UPF1
全体 | 名称: SMG1C-UPF1 complex, comprising SMG1 kinase, SMG8, SMG9 and UPF1 |
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要素 |
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-超分子 #1000: SMG1C-UPF1 complex, comprising SMG1 kinase, SMG8, SMG9 and UPF1
超分子 | 名称: SMG1C-UPF1 complex, comprising SMG1 kinase, SMG8, SMG9 and UPF1 タイプ: sample / ID: 1000 / Number unique components: 4 |
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分子量 | 理論値: 700 KDa |
-分子 #1: Serine/threonine-protein kinase SMG1
分子 | 名称: Serine/threonine-protein kinase SMG1 / タイプ: protein_or_peptide / ID: 1 / Name.synonym: SMG-1 / 組換発現: Yes |
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由来(天然) | 生物種: ![]() |
分子量 | 理論値: 410 KDa |
組換発現 | 生物種: ![]() |
配列 | UniProtKB: Serine/threonine-protein kinase SMG1 GO: DNA repair, RNA metabolic process, nuclear-transcribed mRNA catabolic process, nonsense-mediated decay, ATP binding |
-分子 #2: SMG8
分子 | 名称: SMG8 / タイプ: protein_or_peptide / ID: 2 / Name.synonym: SMG-8 / 組換発現: Yes |
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由来(天然) | 生物種: ![]() |
分子量 | 理論値: 109 KDa |
組換発現 | 生物種: ![]() |
配列 | UniProtKB: Nonsense-mediated mRNA decay factor SMG8 GO: nuclear-transcribed mRNA catabolic process, nonsense-mediated decay InterPro: Nonsense-mediated mRNA decay factor SMG8 |
-分子 #3: SMG9
分子 | 名称: SMG9 / タイプ: protein_or_peptide / ID: 3 / Name.synonym: SMG-9 / 組換発現: Yes |
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由来(天然) | 生物種: ![]() |
分子量 | 理論値: 60 KDa |
組換発現 | 生物種: ![]() |
配列 | UniProtKB: Nonsense-mediated mRNA decay factor SMG9 GO: nuclear-transcribed mRNA catabolic process, nonsense-mediated decay InterPro: P-loop containing nucleoside triphosphate hydrolase, Nonsense-mediated mRNA decay factor SMG8/SMG9 |
-分子 #4: UPF1
分子 | 名称: UPF1 / タイプ: protein_or_peptide / ID: 4 / Name.synonym: RENT1_HUMAN / 組換発現: Yes |
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由来(天然) | 生物種: ![]() |
分子量 | 理論値: 120 KDa |
組換発現 | 生物種: ![]() |
配列 | UniProtKB: Regulator of nonsense transcripts 1 GO: nuclear-transcribed mRNA catabolic process, nonsense-mediated decay InterPro: P-loop containing nucleoside triphosphate hydrolase, RNA helicase UPF1, Cys/His rich zinc-binding domain |
-実験情報
-構造解析
手法 | ネガティブ染色法 |
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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 | 0.01 mg/mL |
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緩衝液 | pH: 7.5 詳細: 10 mM HEPES-KOH, 150 mM NaCl, 20% glycerol, 10 mM MgCl2 |
染色 | タイプ: NEGATIVE / 詳細: 1% uranyl formate |
グリッド | 詳細: 400 mesh grid with thin carbon support, glow discharged |
凍結 | 凍結剤: NONE / 装置: OTHER |
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電子顕微鏡法
顕微鏡 | JEOL 1230 |
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アライメント法 | Legacy - 非点収差: Objective lens astigmatism was corrected using a TVIPS F416 CMOS and the EM-MENU software (TVIPS) |
日付 | 2012年9月27日 |
撮影 | カテゴリ: CCD フィルム・検出器のモデル: TVIPS TEMCAM-F416 (4k x 4k) デジタル化 - サンプリング間隔: 15.6 µm / 実像数: 490 / 平均電子線量: 15 e/Å2 詳細: Using a TVIPS F416 CMOS and the EM-TOOLS software (TVIPS) ビット/ピクセル: 16 |
電子線 | 加速電圧: 100 kV / 電子線源: TUNGSTEN HAIRPIN |
電子光学系 | 倍率(補正後): 54926 / 照射モード: FLOOD BEAM / 撮影モード: BRIGHT FIELD / Cs: 2.9 mm / 最大 デフォーカス(公称値): 2.5 µm / 最小 デフォーカス(公称値): 1.5 µm / 倍率(公称値): 40000 |
試料ステージ | 試料ホルダーモデル: JEOL |
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画像解析
CTF補正 | 詳細: Each micrograph using BSOFT |
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最終 再構成 | 想定した対称性 - 点群: C1 (非対称) / アルゴリズム: OTHER / 解像度のタイプ: BY AUTHOR / 解像度: 20.9 Å / 解像度の算出法: OTHER / ソフトウェア - 名称: EMAN, EMAN2, Xmipp / 使用した粒子像数: 16827 |
最終 2次元分類 | クラス数: 490 |
-原子モデル構築 1
初期モデル | PDB ID: |
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ソフトウェア | 名称: ![]() |
詳細 | The structure was separately fitted using Chimera |
精密化 | 空間: REAL / プロトコル: RIGID BODY FIT / 当てはまり具合の基準: Correlation coefficient |
-原子モデル構築 2
初期モデル | PDB ID: |
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ソフトウェア | 名称: ![]() |
詳細 | HEAT repeat regions from DNA-PKcs were separately fitted into SMG1 using Chimera |
精密化 | 空間: REAL / プロトコル: RIGID BODY FIT / 当てはまり具合の基準: Correlation coefficient |