+データを開く
-基本情報
登録情報 | データベース: EMDB / ID: EMD-18225 | |||||||||
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タイトル | cryo-EM structure of the human spliceosomal B complex protomer (tri-snRNP core region) | |||||||||
マップデータ | ||||||||||
試料 |
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キーワード | spliceosome / pre-catalytic spliceosome / spliceosomal B complex / SPLICING | |||||||||
機能・相同性 | 機能・相同性情報 microfibril / spliceosomal snRNP complex / ribonucleoprotein complex localization / U4atac snRNP / positive regulation of cytotoxic T cell differentiation / maturation of 5S rRNA / RNA localization / U4atac snRNA binding / box C/D sno(s)RNA binding / dense fibrillar component ...microfibril / spliceosomal snRNP complex / ribonucleoprotein complex localization / U4atac snRNP / positive regulation of cytotoxic T cell differentiation / maturation of 5S rRNA / RNA localization / U4atac snRNA binding / box C/D sno(s)RNA binding / dense fibrillar component / box C/D methylation guide snoRNP complex / U4/U6 snRNP / positive regulation of androgen receptor activity / transcription elongation factor activity / snRNP binding / U2-type catalytic step 1 spliceosome / RNA splicing, via transesterification reactions / U4 snRNA binding / spliceosomal tri-snRNP complex / U2-type spliceosomal complex / U2-type precatalytic spliceosome / proline-rich region binding / mRNA cis splicing, via spliceosome / U3 snoRNA binding / U2-type prespliceosome assembly / U2-type catalytic step 2 spliceosome / RNA polymerase binding / box C/D snoRNP assembly / ubiquitin-like protein conjugating enzyme binding / U4 snRNP / U2 snRNP / rRNA modification in the nucleus and cytosol / positive regulation of protein targeting to mitochondrion / U2-type prespliceosome / precatalytic spliceosome / K63-linked polyubiquitin modification-dependent protein binding / mRNA Splicing - Minor Pathway / spliceosomal complex assembly / negative regulation of transcription elongation by RNA polymerase II / mRNA 3'-splice site recognition / MLL1 complex / spliceosomal tri-snRNP complex assembly / single fertilization / U5 snRNA binding / U5 snRNP / Major pathway of rRNA processing in the nucleolus and cytosol / U2 snRNA binding / U6 snRNA binding / ribonucleoprotein complex binding / spliceosomal snRNP assembly / Cajal body / RNA processing / pre-mRNA intronic binding / U1 snRNA binding / U4/U6 x U5 tri-snRNP complex / catalytic step 2 spliceosome / mRNA Splicing - Major Pathway / RNA splicing / maturation of SSU-rRNA / nuclear receptor coactivator activity / response to cocaine / small-subunit processome / nuclear receptor binding / positive regulation of transcription elongation by RNA polymerase II / transcription coregulator activity / spliceosomal complex / protein modification process / mRNA splicing, via spliceosome / mRNA processing / transcription corepressor activity / protein tag activity / ribosomal small subunit biogenesis / cellular response to xenobiotic stimulus / cellular response to tumor necrosis factor / ATPase binding / protein-macromolecule adaptor activity / cellular response to lipopolysaccharide / RNA polymerase II-specific DNA-binding transcription factor binding / transcription coactivator activity / nuclear speck / cell division / intracellular membrane-bounded organelle / GTPase activity / centrosome / chromatin / GTP binding / nucleolus / Golgi apparatus / negative regulation of transcription by RNA polymerase II / positive regulation of transcription by RNA polymerase II / protein-containing complex / DNA binding / RNA binding / zinc ion binding / nucleoplasm / identical protein binding / membrane / nucleus / cytosol / cytoplasm 類似検索 - 分子機能 | |||||||||
生物種 | Homo sapiens (ヒト) | |||||||||
手法 | 単粒子再構成法 / クライオ電子顕微鏡法 / 解像度: 3.1 Å | |||||||||
データ登録者 | Zhang Z / Kumar V / Dybkov O / Will CL / Urlaub H / Stark H / Luehrmann R | |||||||||
資金援助 | ドイツ, 1件
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引用 | ジャーナル: EMBO J / 年: 2024 タイトル: Cryo-EM analyses of dimerized spliceosomes provide new insights into the functions of B complex proteins. 著者: Zhenwei Zhang / Vinay Kumar / Olexandr Dybkov / Cindy L Will / Henning Urlaub / Holger Stark / Reinhard Lührmann / 要旨: The B complex is a key intermediate stage of spliceosome assembly. To improve the structural resolution of monomeric, human spliceosomal B (hB) complexes and thereby generate a more comprehensive hB ...The B complex is a key intermediate stage of spliceosome assembly. To improve the structural resolution of monomeric, human spliceosomal B (hB) complexes and thereby generate a more comprehensive hB molecular model, we determined the cryo-EM structure of B complex dimers formed in the presence of ATP S. The enhanced resolution of these complexes allows a finer molecular dissection of how the 5' splice site (5'ss) is recognized in hB, and new insights into molecular interactions of FBP21, SNU23 and PRP38 with the U6/5'ss helix and with each other. It also reveals that SMU1 and RED are present as a heterotetrameric complex and are located at the interface of the B dimer protomers. We further show that MFAP1 and UBL5 form a 5' exon binding channel in hB, and elucidate the molecular contacts stabilizing the 5' exon at this stage. Our studies thus yield more accurate models of protein and RNA components of hB complexes. They further allow the localization of additional proteins and protein domains (such as SF3B6, BUD31 and TCERG1) whose position was not previously known, thereby uncovering new functions for B-specific and other hB proteins during pre-mRNA splicing. | |||||||||
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_18225.map.gz | 493.2 MB | EMDBマップデータ形式 | |
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ヘッダ (付随情報) | emd-18225-v30.xml emd-18225.xml | 40.4 KB 40.4 KB | 表示 表示 | EMDBヘッダ |
FSC (解像度算出) | emd_18225_fsc.xml | 18.3 KB | 表示 | FSCデータファイル |
画像 | emd_18225.png | 40.6 KB | ||
Filedesc metadata | emd-18225.cif.gz | 13.2 KB | ||
その他 | emd_18225_half_map_1.map.gz emd_18225_half_map_2.map.gz | 428.4 MB 428.4 MB | ||
アーカイブディレクトリ | http://ftp.pdbj.org/pub/emdb/structures/EMD-18225 ftp://ftp.pdbj.org/pub/emdb/structures/EMD-18225 | HTTPS FTP |
-検証レポート
文書・要旨 | emd_18225_validation.pdf.gz | 882.6 KB | 表示 | EMDB検証レポート |
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文書・詳細版 | emd_18225_full_validation.pdf.gz | 882.1 KB | 表示 | |
XML形式データ | emd_18225_validation.xml.gz | 25.6 KB | 表示 | |
CIF形式データ | emd_18225_validation.cif.gz | 34.5 KB | 表示 | |
アーカイブディレクトリ | https://ftp.pdbj.org/pub/emdb/validation_reports/EMD-18225 ftp://ftp.pdbj.org/pub/emdb/validation_reports/EMD-18225 | 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_18225.map.gz / 形式: CCP4 / 大きさ: 536.4 MB / タイプ: IMAGE STORED AS FLOATING POINT NUMBER (4 BYTES) | ||||||||||||||||||||||||||||||||||||
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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: 1.16 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: #2
ファイル | emd_18225_half_map_1.map | ||||||||||||
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投影像・断面図 |
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密度ヒストグラム |
-ハーフマップ: #1
ファイル | emd_18225_half_map_2.map | ||||||||||||
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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 : human pre-catalytic spliceosome
+超分子 #1: human pre-catalytic spliceosome
+分子 #1: U5 snRNA
+分子 #2: U6 snRNA
+分子 #11: MINX pre-mRNA
+分子 #20: U4 snRNA
+分子 #3: Splicing factor 3A subunit 1
+分子 #4: 116 kDa U5 small nuclear ribonucleoprotein component
+分子 #5: Thioredoxin-like protein 4A
+分子 #6: Pre-mRNA-splicing factor 38A
+分子 #7: Microfibrillar-associated protein 1
+分子 #8: NHP2-like protein 1, N-terminally processed
+分子 #9: Protein BUD31 homolog
+分子 #10: WW domain-binding protein 4
+分子 #12: Zinc finger matrin-type protein 2
+分子 #13: Ubiquitin-like protein 5
+分子 #14: U4/U6 small nuclear ribonucleoprotein Prp31
+分子 #15: U4/U6 small nuclear ribonucleoprotein Prp4
+分子 #16: Pre-mRNA-processing factor 6
+分子 #17: Pre-mRNA-processing-splicing factor 8
+分子 #18: U4/U6.U5 tri-snRNP-associated protein 1
+分子 #19: Transcription elongation regulator 1
+分子 #21: U4/U6 small nuclear ribonucleoprotein Prp3
-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
緩衝液 | pH: 7.9 |
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凍結 | 凍結剤: ETHANE |
-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: FEI FALCON III (4k x 4k) 検出モード: INTEGRATING / 平均電子線量: 48.0 e/Å2 |
電子線 | 加速電圧: 300 kV / 電子線源: FIELD EMISSION GUN |
電子光学系 | 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): 3.0 µm / 最小 デフォーカス(公称値): 1.5 µm |
実験機器 | モデル: Titan Krios / 画像提供: FEI Company |