+
データを開く
-
基本情報
登録情報 | データベース: EMDB / ID: EMD-3278 | |||||||||
---|---|---|---|---|---|---|---|---|---|---|
タイトル | Electron microscopy of the SMG1C-DHX34 complex | |||||||||
![]() | Reconstruction of SMG1C-DHX34 | |||||||||
![]() |
| |||||||||
![]() | NMD / SMG1 / SMG8 / SMG9 / PIKK / DHX34 / RNA degradation / RNA helicase | |||||||||
機能・相同性 | ![]() RNA helicase activity => GO:0003724 / RNA binding => GO:0003723 / negative regulation of nuclear-transcribed mRNA catabolic process, nonsense-mediated decay / RNA metabolic process / diacylglycerol-dependent serine/threonine kinase activity / chromatoid body / eye development / regulation of protein kinase activity / nuclear-transcribed mRNA catabolic process, nonsense-mediated decay / regulation of telomere maintenance ...RNA helicase activity => GO:0003724 / RNA binding => GO:0003723 / negative regulation of nuclear-transcribed mRNA catabolic process, nonsense-mediated decay / RNA metabolic process / diacylglycerol-dependent serine/threonine kinase activity / chromatoid body / eye development / regulation of protein kinase activity / nuclear-transcribed mRNA catabolic process, nonsense-mediated decay / regulation of telomere maintenance / nuclear-transcribed mRNA catabolic process / telomeric DNA binding / phosphatidylinositol phosphate biosynthetic process / Nonsense Mediated Decay (NMD) enhanced by the Exon Junction Complex (EJC) / RNA processing / mRNA export from nucleus / positive regulation of phosphorylation / brain development / heart development / peptidyl-serine phosphorylation / in utero embryonic development / protein autophosphorylation / RNA helicase activity / non-specific serine/threonine protein kinase / protein kinase activity / RNA helicase / DNA repair / protein serine kinase activity / protein serine/threonine kinase activity / DNA damage response / protein-containing complex binding / negative regulation of apoptotic process / ATP hydrolysis activity / RNA binding / nucleoplasm / ATP binding / identical protein binding / membrane / nucleus / metal ion binding / cytoplasm / cytosol 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: 20.89 Å | |||||||||
![]() | Melero R / Hug N / Lopez-Perrote A / Yamashita A / Caceres J / Llorca O | |||||||||
![]() | ![]() タイトル: The RNA helicase DHX34 functions as a scaffold for SMG1-mediated UPF1 phosphorylation. 著者: Roberto Melero / Nele Hug / Andrés López-Perrote / Akio Yamashita / Javier F Cáceres / Oscar Llorca / ![]() ![]() ![]() 要旨: Nonsense-mediated decay (NMD) is a messenger RNA quality-control pathway triggered by SMG1-mediated phosphorylation of the NMD factor UPF1. In recent times, the RNA helicase DHX34 was found to ...Nonsense-mediated decay (NMD) is a messenger RNA quality-control pathway triggered by SMG1-mediated phosphorylation of the NMD factor UPF1. In recent times, the RNA helicase DHX34 was found to promote mRNP remodelling, leading to activation of NMD. Here we demonstrate the mechanism by which DHX34 functions in concert with SMG1. DHX34 comprises two distinct structural units, a core that binds UPF1 and a protruding carboxy-terminal domain (CTD) that binds the SMG1 kinase, as shown using truncated forms of DHX34 and electron microscopy of the SMG1-DHX34 complex. Truncation of the DHX34 CTD does not affect binding to UPF1; however, it compromises DHX34 binding to SMG1 to affect UPF1 phosphorylation and hence abrogate NMD. Altogether, these data suggest the existence of a complex comprising SMG1, UPF1 and DHX34, with DHX34 functioning as a scaffold for UPF1 and SMG1. This complex promotes UPF1 phosphorylation leading to functional NMD. | |||||||||
履歴 |
|
-
構造の表示
ムービー |
![]() |
---|---|
構造ビューア | EMマップ: ![]() ![]() ![]() |
添付画像 |
-
ダウンロードとリンク
-EMDBアーカイブ
マップデータ | ![]() | 749.1 KB | ![]() | |
---|---|---|---|---|
ヘッダ (付随情報) | ![]() ![]() | 13.6 KB 13.6 KB | 表示 表示 | ![]() |
画像 | ![]() | 104.5 KB | ||
アーカイブディレクトリ | ![]() ![]() | HTTPS FTP |
-検証レポート
文書・要旨 | ![]() | 207.8 KB | 表示 | ![]() |
---|---|---|---|---|
文書・詳細版 | ![]() | 206.9 KB | 表示 | |
XML形式データ | ![]() | 4.9 KB | 表示 | |
アーカイブディレクトリ | ![]() ![]() | HTTPS FTP |
-関連構造データ
-
リンク
EMDBのページ | ![]() ![]() |
---|---|
「今月の分子」の関連する項目 |
-
マップ
ファイル | ![]() | ||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|
注釈 | Reconstruction of SMG1C-DHX34 | ||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
ボクセルのサイズ | X=Y=Z: 5.68 Å | ||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
密度 |
| ||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
対称性 | 空間群: 1 | ||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
詳細 | EMDB XML:
CCP4マップ ヘッダ情報:
|
-添付データ
-
試料の構成要素
-全体 : SMG1C complex, comprising SMG1 kinase, SMG8 and SMG9, bound to th...
全体 | 名称: SMG1C complex, comprising SMG1 kinase, SMG8 and SMG9, bound to the DHX34 helicase |
---|---|
要素 |
|
-超分子 #1000: SMG1C complex, comprising SMG1 kinase, SMG8 and SMG9, bound to th...
超分子 | 名称: SMG1C complex, comprising SMG1 kinase, SMG8 and SMG9, bound to the DHX34 helicase タイプ: sample / ID: 1000 / 集合状態: Heterotetramer of SMG1, SMG8, SMG9 and DHX34 / Number unique components: 4 |
---|---|
分子量 | 理論値: 770 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 |
---|---|
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 |
---|---|
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 |
---|---|
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: DHX34
分子 | 名称: DHX34 / タイプ: protein_or_peptide / ID: 4 / 組換発現: Yes |
---|---|
由来(天然) | 生物種: ![]() |
分子量 | 理論値: 128 KDa |
組換発現 | 生物種: ![]() |
配列 | UniProtKB: Probable ATP-dependent RNA helicase DHX34 GO: cytoplasm, membrane, ATP binding, RNA helicase activity => GO:0003724, RNA binding => GO:0003723, negative regulation of nuclear-transcribed mRNA catabolic process, nonsense-mediated decay, ...GO: cytoplasm, membrane, ATP binding, RNA helicase activity => GO:0003724, RNA binding => GO:0003723, negative regulation of nuclear-transcribed mRNA catabolic process, nonsense-mediated decay, nuclear-transcribed mRNA catabolic process, nuclear-transcribed mRNA catabolic process, nonsense-mediated decay, RNA processing InterPro: DEAD-box helicase, OB fold, Helicase-associated domain, Helicase superfamily 1/2, ATP-binding domain, Helicase, C-terminal, P-loop containing nucleoside triphosphate hydrolase, INTERPRO: IPR015880 |
-実験情報
-構造解析
手法 | ネガティブ染色法 |
---|---|
![]() | 単粒子再構成法 |
試料の集合状態 | particle |
-
試料調製
濃度 | 0.01 mg/mL |
---|---|
緩衝液 | pH: 7.5 詳細: 10 mM HEPES-KOH, 150 mM NaCl, 20% (v/v) 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 |
-
電子顕微鏡法
顕微鏡 | JEOL 1230 |
---|---|
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 | 2013年7月25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: 542 / 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 |
-
画像解析
CTF補正 | 詳細: Each micrograph using BSOFT |
---|---|
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: 20.89 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: 21020 |
最終 2次元分類 | クラス数: 490 |