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基本情報
登録情報 | データベース: EMDB / ID: EMD-7808 | |||||||||
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タイトル | Human nuclear exosome-MTR4 RNA complex - overall reconstruction | |||||||||
![]() | Overall reconstruction for human nuclear exosome-MTR4 helicase complex | |||||||||
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![]() | RNA exosome / RNA degradation / ribonuclease / helicase / SF2 / RNA-protein complex / translocase / nuclear / HYDROLASE | |||||||||
機能・相同性 | ![]() DNA deamination / nucleolar exosome (RNase complex) / snRNA catabolic process / positive regulation of mRNA cis splicing, via spliceosome / nuclear polyadenylation-dependent snoRNA catabolic process / nuclear polyadenylation-dependent snRNA catabolic process / nuclear polyadenylation-dependent antisense transcript catabolic process / TRAMP complex / RNA exonuclease activity / nuclear mRNA surveillance of mRNA 3'-end processing ...DNA deamination / nucleolar exosome (RNase complex) / snRNA catabolic process / positive regulation of mRNA cis splicing, via spliceosome / nuclear polyadenylation-dependent snoRNA catabolic process / nuclear polyadenylation-dependent snRNA catabolic process / nuclear polyadenylation-dependent antisense transcript catabolic process / TRAMP complex / RNA exonuclease activity / nuclear mRNA surveillance of mRNA 3'-end processing / regulation of telomerase RNA localization to Cajal body / U1 snRNA 3'-end processing / nuclear polyadenylation-dependent CUT catabolic process / U5 snRNA 3'-end processing / TRAMP-dependent tRNA surveillance pathway / CUT catabolic process / exosome (RNase complex) / U4 snRNA 3'-end processing / cytoplasmic exosome (RNase complex) / mRNA decay by 3' to 5' exoribonuclease / nuclear polyadenylation-dependent rRNA catabolic process / poly(A)-dependent snoRNA 3'-end processing / nuclear exosome (RNase complex) / exonucleolytic trimming to generate mature 3'-end of 5.8S rRNA from tricistronic rRNA transcript (SSU-rRNA, 5.8S rRNA, LSU-rRNA) / Tristetraprolin (TTP, ZFP36) binds and destabilizes mRNA / ATF4 activates genes in response to endoplasmic reticulum stress / histone mRNA catabolic process / Butyrate Response Factor 1 (BRF1) binds and destabilizes mRNA / positive regulation of isotype switching / nuclear mRNA surveillance / rRNA catabolic process / 7S RNA binding / mRNA 3'-UTR AU-rich region binding / isotype switching / telomerase RNA binding / 加水分解酵素; エステル加水分解酵素; 5'-リン酸モノエステル産生エンドリボヌクレアーゼ / RNA catabolic process / nuclear-transcribed mRNA catabolic process, deadenylation-dependent decay / nuclear-transcribed mRNA catabolic process, nonsense-mediated decay / KSRP (KHSRP) binds and destabilizes mRNA / maturation of 5.8S rRNA / nuclear chromosome / negative regulation of telomere maintenance via telomerase / mRNA catabolic process / nuclear-transcribed mRNA catabolic process / Major pathway of rRNA processing in the nucleolus and cytosol / RNA processing / catalytic step 2 spliceosome / mRNA Splicing - Major Pathway / Regulation of endogenous retroelements by the Human Silencing Hub (HUSH) complex / guanyl-nucleotide exchange factor activity / small-subunit processome / euchromatin / mRNA splicing, via spliceosome / fibrillar center / rRNA processing / chromosome / ribosomal small subunit biogenesis / 加水分解酵素; エステル加水分解酵素; 5'-リン酸モノエステル産生エキソリボヌクレアーゼ / positive regulation of cell growth / 3'-5'-RNA exonuclease activity / endonuclease activity / defense response to virus / RNA polymerase II-specific DNA-binding transcription factor binding / single-stranded RNA binding / RNA helicase activity / nuclear speck / immune response / RNA helicase / DNA repair / nucleotide binding / intracellular membrane-bounded organelle / DNA damage response / nucleolus / positive regulation of transcription by RNA polymerase II / ATP hydrolysis activity / DNA binding / RNA binding / extracellular exosome / nucleoplasm / ATP binding / metal ion 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: 3.45 Å | |||||||||
![]() | Weick E-M / Lima CD | |||||||||
資金援助 | ![]()
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![]() | ![]() タイトル: Helicase-Dependent RNA Decay Illuminated by a Cryo-EM Structure of a Human Nuclear RNA Exosome-MTR4 Complex. 著者: Eva-Maria Weick / M Rhyan Puno / Kurt Januszyk / John C Zinder / Michael A DiMattia / Christopher D Lima / ![]() 要旨: The ribonucleolytic RNA exosome interacts with RNA helicases to degrade RNA. To understand how the 3' to 5' Mtr4 helicase engages RNA and the nuclear exosome, we reconstituted 14-subunit Mtr4- ...The ribonucleolytic RNA exosome interacts with RNA helicases to degrade RNA. To understand how the 3' to 5' Mtr4 helicase engages RNA and the nuclear exosome, we reconstituted 14-subunit Mtr4-containing RNA exosomes from Saccharomyces cerevisiae, Schizosaccharomyces pombe, and human and show that they unwind structured substrates to promote degradation. We loaded a human exosome with an optimized DNA-RNA chimera that stalls MTR4 during unwinding and determined its structure to an overall resolution of 3.45 Å by cryoelectron microscopy (cryo-EM). The structure reveals an RNA-engaged helicase atop the non-catalytic core, with RNA captured within the central channel and DIS3 exoribonuclease active site. MPP6 tethers MTR4 to the exosome through contacts to the RecA domains of MTR4. EXOSC10 remains bound to the core, but its catalytic module and cofactor C1D are displaced by RNA-engaged MTR4. Competition for the exosome core may ensure that RNA is committed to degradation by DIS3 when engaged by MTR4. | |||||||||
履歴 |
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構造の表示
ムービー |
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構造ビューア | EMマップ: ![]() ![]() ![]() |
添付画像 |
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マップデータ | ![]() | 15.9 MB | ![]() | |
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ヘッダ (付随情報) | ![]() ![]() | 36.2 KB 36.2 KB | 表示 表示 | ![]() |
FSC (解像度算出) | ![]() | 13.6 KB | 表示 | ![]() |
画像 | ![]() | 46.2 KB | ||
Filedesc metadata | ![]() | 11 KB | ||
アーカイブディレクトリ | ![]() ![]() | HTTPS FTP |
-関連構造データ
関連構造データ | ![]() 6d6qMC ![]() 7809C ![]() 7810C ![]() 7812C ![]() 7813C ![]() 7814C ![]() 7815C ![]() 7818C ![]() 7819C ![]() 6d6rC C: 同じ文献を引用 ( M: このマップから作成された原子モデル |
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類似構造データ |
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リンク
EMDBのページ | ![]() ![]() |
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「今月の分子」の関連する項目 |
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マップ
ファイル | ![]() | ||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
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注釈 | Overall reconstruction for human nuclear exosome-MTR4 helicase complex | ||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
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: 1.07 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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-添付データ
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試料の構成要素
+全体 : Human nuclear exosome-MTR4 helicase captured after unwinding a DN...
+超分子 #1: Human nuclear exosome-MTR4 helicase captured after unwinding a DN...
+分子 #1: Exosome complex component RRP45
+分子 #2: Exosome complex component RRP41
+分子 #3: Exosome complex component RRP43
+分子 #4: Exosome complex component RRP46
+分子 #5: Exosome complex component RRP42
+分子 #6: Exosome complex component MTR3
+分子 #7: Exosome complex component RRP40
+分子 #8: Exosome complex component RRP4
+分子 #9: Exosome complex component CSL4
+分子 #10: Exosome component 10
+分子 #11: Exosome complex exonuclease RRP44
+分子 #12: M-phase phosphoprotein 6
+分子 #13: Exosome RNA helicase MTR4
+分子 #14: RNA (5'-R(*AP*GP*CP*AP*CP*CP*GP*UP*AP*AP*AP*GP*AP*CP*GP*C)-3')
+分子 #15: DNA/RNA (62-MER)
+分子 #16: MAGNESIUM ION
+分子 #17: ZINC ION
+分子 #18: PHOSPHOAMINOPHOSPHONIC ACID-ADENYLATE ESTER
-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 | 1.0 mg/mL | |||||||||||||||||||||
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緩衝液 | pH: 8 構成要素:
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グリッド | モデル: Quantifoil R2/2 / 材質: GOLD / 前処理 - タイプ: GLOW DISCHARGE / 前処理 - 時間: 60 sec. / 前処理 - 雰囲気: AIR / 前処理 - 気圧: 0.037 kPa | |||||||||||||||||||||
凍結 | 凍結剤: ETHANE / チャンバー内温度: 298 K / 装置: FEI VITROBOT MARK IV / 詳細: 30 sec wait time, 2.5 sec blot time. | |||||||||||||||||||||
詳細 | Sample was monodisperse upon elution from gel filtration prior to vitrification. |
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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: SUPER-RESOLUTION / 実像数: 1439 / 平均電子線量: 85.23 e/Å2 |
電子線 | 加速電圧: 300 kV / 電子線源: ![]() |
電子光学系 | 照射モード: FLOOD BEAM / 撮影モード: BRIGHT FIELD |
実験機器 | ![]() モデル: Titan Krios / 画像提供: FEI Company |
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画像解析
-原子モデル構築 1
詳細 | Models were rebuilt manually using Coot, with real space refinement with local scaling followed by Phenix real space refinement with suboptimal global scaling. |
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精密化 | 空間: REAL / プロトコル: BACKBONE TRACE / 温度因子: 73.6 / 当てはまり具合の基準: Correlation coefficient |
得られたモデル | ![]() PDB-6d6q: |