+データを開く
-基本情報
登録情報 | データベース: EMDB / ID: EMD-26941 | ||||||||||||
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タイトル | Nucleoplasmic pre-60S intermediate of the Nog2 containing pre-rotation state from a Spb1 D52A suppressor 3 strain | ||||||||||||
マップデータ | Nucleoplasmic pre-60S intermediate of the Nog2 containing pre-rotation state from a Spb1 D52A suppressor 3 strain | ||||||||||||
試料 |
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キーワード | ribosome biogenesis / K-loop GTPase / GTPase / RIBOSOME | ||||||||||||
機能・相同性 | 機能・相同性情報 regulation of ribosomal subunit export from nucleus / nuclear exosome (RNase complex) / 加水分解酵素 / PeBoW complex / maturation of 5.8S rRNA from tricistronic rRNA transcript (SSU-rRNA, 5.8S rRNA, LSU-rRNA) / maturation of 5.8S rRNA / hexon binding / pre-mRNA 5'-splice site binding / cleavage in ITS2 between 5.8S rRNA and LSU-rRNA of tricistronic rRNA transcript (SSU-rRNA, 5.8S rRNA, LSU-rRNA) / proteasome binding ...regulation of ribosomal subunit export from nucleus / nuclear exosome (RNase complex) / 加水分解酵素 / PeBoW complex / maturation of 5.8S rRNA from tricistronic rRNA transcript (SSU-rRNA, 5.8S rRNA, LSU-rRNA) / maturation of 5.8S rRNA / hexon binding / pre-mRNA 5'-splice site binding / cleavage in ITS2 between 5.8S rRNA and LSU-rRNA of tricistronic rRNA transcript (SSU-rRNA, 5.8S rRNA, LSU-rRNA) / proteasome binding / Major pathway of rRNA processing in the nucleolus and cytosol / SRP-dependent cotranslational protein targeting to membrane / GTP hydrolysis and joining of the 60S ribosomal subunit / ribosomal large subunit binding / Nonsense Mediated Decay (NMD) independent of the Exon Junction Complex (EJC) / Nonsense Mediated Decay (NMD) enhanced by the Exon Junction Complex (EJC) / Formation of a pool of free 40S subunits / negative regulation of mRNA splicing, via spliceosome / preribosome, large subunit precursor / nuclear-transcribed mRNA catabolic process / L13a-mediated translational silencing of Ceruloplasmin expression / translational elongation / ribosomal large subunit export from nucleus / regulation of translational fidelity / protein-RNA complex assembly / ribonucleoprotein complex binding / ribosomal subunit export from nucleus / endonucleolytic cleavage in ITS1 to separate SSU-rRNA from 5.8S rRNA and LSU-rRNA from tricistronic rRNA transcript (SSU-rRNA, 5.8S rRNA, LSU-rRNA) / maturation of LSU-rRNA / Neutrophil degranulation / translation initiation factor activity / maturation of LSU-rRNA from tricistronic rRNA transcript (SSU-rRNA, 5.8S rRNA, LSU-rRNA) / proteasome complex / assembly of large subunit precursor of preribosome / ribosomal large subunit biogenesis / maturation of SSU-rRNA from tricistronic rRNA transcript (SSU-rRNA, 5.8S rRNA, LSU-rRNA) / maturation of SSU-rRNA / cytosolic ribosome assembly / small-subunit processome / macroautophagy / maintenance of translational fidelity / protein catabolic process / rRNA processing / metallopeptidase activity / protein transport / ribosome biogenesis / viral capsid / protein-macromolecule adaptor activity / 5S rRNA binding / large ribosomal subunit rRNA binding / ribosomal large subunit assembly / cytoplasmic translation / cytosolic large ribosomal subunit / nucleic acid binding / negative regulation of translation / rRNA binding / ribosome / structural constituent of ribosome / translation / GTPase activity / mRNA binding / host cell nucleus / GTP binding / nucleolus / proteolysis / RNA binding / zinc ion binding / nucleoplasm / identical protein binding / nucleus / metal ion binding / cytosol / cytoplasm 類似検索 - 分子機能 | ||||||||||||
生物種 | Saccharomyces cerevisiae BY4741 (パン酵母) | ||||||||||||
手法 | 単粒子再構成法 / クライオ電子顕微鏡法 / 解像度: 2.36 Å | ||||||||||||
データ登録者 | Sekulski K / Cruz VE / Weirich CS / Erzberger JP | ||||||||||||
資金援助 | 米国, 3件
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引用 | ジャーナル: Nat Commun / 年: 2023 タイトル: rRNA methylation by Spb1 regulates the GTPase activity of Nog2 during 60S ribosomal subunit assembly. 著者: Kamil Sekulski / Victor Emmanuel Cruz / Christine S Weirich / Jan P Erzberger / 要旨: Biogenesis of the large ribosomal (60S) subunit involves the assembly of three rRNAs and 46 proteins, a process requiring approximately 70 ribosome biogenesis factors (RBFs) that bind and release the ...Biogenesis of the large ribosomal (60S) subunit involves the assembly of three rRNAs and 46 proteins, a process requiring approximately 70 ribosome biogenesis factors (RBFs) that bind and release the pre-60S at specific steps along the assembly pathway. The methyltransferase Spb1 and the K-loop GTPase Nog2 are essential RBFs that engage the rRNA A-loop during sequential steps in 60S maturation. Spb1 methylates the A-loop nucleotide G2922 and a catalytically deficient mutant strain (spb1) has a severe 60S biogenesis defect. However, the assembly function of this modification is currently unknown. Here, we present cryo-EM reconstructions that reveal that unmethylated G2922 leads to the premature activation of Nog2 GTPase activity and capture a Nog2-GDP-AlF transition state structure that implicates the direct involvement of unmodified G2922 in Nog2 GTPase activation. Genetic suppressors and in vivo imaging indicate that premature GTP hydrolysis prevents the efficient binding of Nog2 to early nucleoplasmic 60S intermediates. We propose that G2922 methylation levels regulate Nog2 recruitment to the pre-60S near the nucleolar/nucleoplasmic phase boundary, forming a kinetic checkpoint to regulate 60S production. Our approach and findings provide a template to study the GTPase cycles and regulatory factor interactions of the other K-loop GTPases involved in ribosome assembly. | ||||||||||||
履歴 |
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-構造の表示
添付画像 |
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-ダウンロードとリンク
-EMDBアーカイブ
マップデータ | emd_26941.map.gz | 117.2 MB | EMDBマップデータ形式 | |
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ヘッダ (付随情報) | emd-26941-v30.xml emd-26941.xml | 91.1 KB 91.1 KB | 表示 表示 | EMDBヘッダ |
FSC (解像度算出) | emd_26941_fsc.xml | 11.3 KB | 表示 | FSCデータファイル |
画像 | emd_26941.png | 104.8 KB | ||
Filedesc metadata | emd-26941.cif.gz | 19.1 KB | ||
その他 | emd_26941_half_map_1.map.gz emd_26941_half_map_2.map.gz | 98.4 MB 98.5 MB | ||
アーカイブディレクトリ | http://ftp.pdbj.org/pub/emdb/structures/EMD-26941 ftp://ftp.pdbj.org/pub/emdb/structures/EMD-26941 | HTTPS FTP |
-検証レポート
文書・要旨 | emd_26941_validation.pdf.gz | 1.1 MB | 表示 | EMDB検証レポート |
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文書・詳細版 | emd_26941_full_validation.pdf.gz | 1.1 MB | 表示 | |
XML形式データ | emd_26941_validation.xml.gz | 18.6 KB | 表示 | |
CIF形式データ | emd_26941_validation.cif.gz | 24.4 KB | 表示 | |
アーカイブディレクトリ | https://ftp.pdbj.org/pub/emdb/validation_reports/EMD-26941 ftp://ftp.pdbj.org/pub/emdb/validation_reports/EMD-26941 | HTTPS FTP |
-関連構造データ
-リンク
EMDBのページ | EMDB (EBI/PDBe) / EMDataResource |
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「今月の分子」の関連する項目 |
-マップ
ファイル | ダウンロード / ファイル: emd_26941.map.gz / 形式: CCP4 / 大きさ: 125 MB / タイプ: IMAGE STORED AS FLOATING POINT NUMBER (4 BYTES) | ||||||||||||||||||||||||||||||||||||
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注釈 | Nucleoplasmic pre-60S intermediate of the Nog2 containing pre-rotation state from a Spb1 D52A suppressor 3 strain | ||||||||||||||||||||||||||||||||||||
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: 1.08 Å | ||||||||||||||||||||||||||||||||||||
密度 |
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対称性 | 空間群: 1 | ||||||||||||||||||||||||||||||||||||
詳細 | EMDB XML:
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-添付データ
-ハーフマップ: Half-map 1 of nucleoplasmic pre-60S intermediate of the...
ファイル | emd_26941_half_map_1.map | ||||||||||||
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注釈 | Half-map 1 of nucleoplasmic pre-60S intermediate of the Nog2 containing pre-rotation state from a Spb1 D52A suppressor 3 strain | ||||||||||||
投影像・断面図 |
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密度ヒストグラム |
-ハーフマップ: Half-map 2 of nucleoplasmic pre-60S intermediate of the...
ファイル | emd_26941_half_map_2.map | ||||||||||||
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注釈 | Half-map 2 of nucleoplasmic pre-60S intermediate of the Nog2 containing pre-rotation state from a Spb1 D52A suppressor 3 strain | ||||||||||||
投影像・断面図 |
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密度ヒストグラム |
-試料の構成要素
+全体 : Nucleolar 60S intermediate purified with tags on Tif6 and Nog2.
+超分子 #1: Nucleolar 60S intermediate purified with tags on Tif6 and Nog2.
+分子 #1: 60S ribosomal protein L2-A
+分子 #5: rRNA-processing protein
+分子 #7: 60S ribosomal protein L12-A
+分子 #8: Ribosome biogenesis protein ALB1
+分子 #9: 60S ribosomal protein L3
+分子 #10: 60S ribosomal protein L4-A
+分子 #11: 60S ribosomal protein L5
+分子 #12: 60S ribosomal protein L6-A
+分子 #13: 60S ribosomal protein L7-A
+分子 #14: RPL8A isoform 1
+分子 #15: 60S ribosomal protein L9-A
+分子 #16: Bud site selection protein 20
+分子 #17: 60S ribosomal protein L11-A
+分子 #18: Proteasome-interacting protein CIC1
+分子 #19: 60S ribosomal protein L13-A
+分子 #20: 60S ribosomal protein L14-A
+分子 #21: 60S ribosomal protein L15-A
+分子 #22: 60S ribosomal protein L16-A
+分子 #23: 60S ribosomal protein L17-A
+分子 #24: 60S ribosomal protein L18-A
+分子 #25: 60S ribosomal protein L19-A
+分子 #26: 60S ribosomal protein L20-A
+分子 #27: 60S ribosomal protein L21-A
+分子 #28: 60S ribosomal protein L22-A
+分子 #29: 60S ribosomal protein L23-A
+分子 #30: Ribosome assembly factor MRT4
+分子 #31: 60S ribosomal protein L25
+分子 #32: 60S ribosomal protein L26-A
+分子 #33: 60S ribosomal protein L27-A
+分子 #34: 60S ribosomal protein L28
+分子 #35: Nucleolar GTP-binding protein 1
+分子 #36: 60S ribosomal protein L30
+分子 #37: 60S ribosomal protein L31-A
+分子 #38: 60S ribosomal protein L32
+分子 #39: 60S ribosomal protein L33-A
+分子 #40: 60S ribosomal protein L34-A
+分子 #41: 60S ribosomal protein L35-A
+分子 #42: 60S ribosomal protein L36-A
+分子 #43: 60S ribosomal protein L37-A
+分子 #44: RPL38 isoform 1
+分子 #45: 60S ribosomal protein L39
+分子 #46: Nucleolar GTP-binding protein 2
+分子 #47: Pescadillo homolog
+分子 #48: Ribosome biogenesis protein 15
+分子 #49: 60S ribosomal protein L43-A
+分子 #50: Ribosome biogenesis protein NOP53
+分子 #51: Ribosome biogenesis protein NSA2
+分子 #52: Nuclear GTP-binding protein NUG1
+分子 #53: RLP7 isoform 1
+分子 #54: Ribosome biogenesis protein RLP24
+分子 #55: Ribosome production factor 2 homolog
+分子 #56: Ribosome biogenesis regulatory protein
+分子 #57: RSA4 isoform 1
+分子 #58: Eukaryotic translation initiation factor 6
+分子 #59: UPF0642 protein YBL028C
+分子 #60: Probable metalloprotease ARX1
+分子 #2: 25S rRNA
+分子 #3: 5.8S rRNA
+分子 #4: 5S rRNA
+分子 #6: ITS2
+分子 #61: MAGNESIUM ION
+分子 #62: 2-[3-(2-HYDROXY-1,1-DIHYDROXYMETHYL-ETHYLAMINO)-PROPYLAMINO]-2-HY...
+分子 #63: ZINC ION
+分子 #64: GUANOSINE-5'-DIPHOSPHATE
+分子 #65: POTASSIUM ION
+分子 #66: water
-実験情報
-構造解析
手法 | クライオ電子顕微鏡法 |
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解析 | 単粒子再構成法 |
試料の集合状態 | particle |
-試料調製
濃度 | 0.8 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 R2/1 / 材質: COPPER / メッシュ: 300 / 支持フィルム - 材質: CARBON / 支持フィルム - トポロジー: CONTINUOUS / 支持フィルム - Film thickness: 0.3 | |||||||||||||||||||||
凍結 | 凍結剤: ETHANE / チャンバー内湿度: 100 % / チャンバー内温度: 277 K / 装置: FEI VITROBOT MARK IV |
-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 K3 (6k x 4k) / 撮影したグリッド数: 1 / 実像数: 4349 / 平均露光時間: 0.05 sec. / 平均電子線量: 1.4 e/Å2 |
電子線 | 加速電圧: 300 kV / 電子線源: FIELD EMISSION GUN |
電子光学系 | C2レンズ絞り径: 70.0 µm / 最大 デフォーカス(補正後): 2.2 µm / 最小 デフォーカス(補正後): 0.9 µm / 照射モード: FLOOD BEAM / 撮影モード: BRIGHT FIELD / 最大 デフォーカス(公称値): 2.2 µm / 最小 デフォーカス(公称値): 0.9 µm / 倍率(公称値): 81000 |
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 |