+データを開く
-基本情報
登録情報 | データベース: PDB / ID: 6qx9 | |||||||||
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タイトル | Structure of a human fully-assembled precatalytic spliceosome (pre-B complex). | |||||||||
要素 |
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キーワード | SPLICING / RNP complex / RNA (リボ核酸) / protein (タンパク質) / spliceosome (スプライセオソーム) | |||||||||
機能・相同性 | 機能・相同性情報 mRNA Splicing - Minor Pathway / spliceosomal snRNP complex / negative regulation of protein refolding / Lsm2-8 complex / U6 snRNA 3'-end binding / ribonucleoprotein complex localization / U4atac snRNP / positive regulation of cytotoxic T cell differentiation / maturation of 5S rRNA / R-loop processing ...mRNA Splicing - Minor Pathway / spliceosomal snRNP complex / negative regulation of protein refolding / Lsm2-8 complex / U6 snRNA 3'-end binding / ribonucleoprotein complex localization / U4atac snRNP / positive regulation of cytotoxic T cell differentiation / maturation of 5S rRNA / R-loop processing / RNA localization / mRNA Splicing - Major Pathway / U4atac snRNA binding / mRNA decay by 5' to 3' exoribonuclease / Lsm1-7-Pat1 complex / U11/U12 snRNP / box C/D sno(s)RNA binding / snRNP binding / U6 snRNP / regulation of ATP-dependent activity / PH domain binding / U2 snRNP binding / U7 snRNA binding / histone pre-mRNA DCP binding / 核小体 / U7 snRNP / B-WICH complex / histone pre-mRNA 3'end processing complex / cis assembly of pre-catalytic spliceosome / SLBP independent Processing of Histone Pre-mRNAs / SLBP Dependent Processing of Replication-Dependent Histone Pre-mRNAs / spliceosome conformational change to release U4 (or U4atac) and U1 (or U11) / box C/D methylation guide snoRNP complex / splicing factor binding / U4/U6 snRNP / protein methylation / U12-type spliceosomal complex / methylosome / P-body assembly / 7-methylguanosine cap hypermethylation / U2-type catalytic step 1 spliceosome / pICln-Sm protein complex / U1 snRNP binding / RNA splicing, via transesterification reactions / spliceosomal tri-snRNP complex / small nuclear ribonucleoprotein complex / P granule / sno(s)RNA-containing ribonucleoprotein complex / U4 snRNA binding / SMN-Sm protein complex / : / mRNA cis splicing, via spliceosome / U2-type spliceosomal complex / telomerase RNA binding / telomerase holoenzyme complex / positive regulation of mRNA splicing, via spliceosome / U2-type precatalytic spliceosome / negative regulation of chaperone-mediated autophagy / U2-type prespliceosome assembly / box C/D snoRNP assembly / commitment complex / U2-type catalytic step 2 spliceosome / U4 snRNP / rRNA modification in the nucleus and cytosol / U2 snRNP / SAGA complex / nuclear-transcribed mRNA catabolic process / RNA Polymerase II Transcription Termination / positive regulation of transcription by RNA polymerase III / U1 snRNP / U3 snoRNA binding / U2-type prespliceosome / K63-linked polyubiquitin modification-dependent protein binding / tRNA processing / pre-mRNA 5'-splice site binding / precatalytic spliceosome / nuclear androgen receptor binding / spliceosomal complex assembly / positive regulation of transcription by RNA polymerase I / mRNA Splicing - Minor Pathway / regulation of RNA splicing / single fertilization / mRNA catabolic process / negative regulation of mRNA splicing, via spliceosome / MLL1 complex / mRNA 3'-splice site recognition / mRNA 5'-splice site recognition / spliceosomal tri-snRNP complex assembly / Major pathway of rRNA processing in the nucleolus and cytosol / U5 snRNA binding / U5 snRNP / regulation of DNA repair / カハール体 / U2 snRNA binding / spliceosomal snRNP assembly / U6 snRNA binding / 転写後修飾 / ribonucleoprotein complex binding / pre-mRNA intronic binding / maturation of SSU-rRNA 類似検索 - 分子機能 | |||||||||
生物種 | Homo sapiens (ヒト) Human adenovirus 2 (ヒトアデノウイルス) | |||||||||
手法 | 電子顕微鏡法 / 単粒子再構成法 / クライオ電子顕微鏡法 / 解像度: 3.28 Å | |||||||||
データ登録者 | Charenton, C. / Wilkinson, M.E. / Nagai, K. | |||||||||
資金援助 | 英国, 2件
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引用 | ジャーナル: Science / 年: 2019 タイトル: Mechanism of 5' splice site transfer for human spliceosome activation. 著者: Clément Charenton / Max E Wilkinson / Kiyoshi Nagai / 要旨: The prespliceosome, comprising U1 and U2 small nuclear ribonucleoproteins (snRNPs) bound to the precursor messenger RNA 5' splice site (5'SS) and branch point sequence, associates with the U4/U6.U5 ...The prespliceosome, comprising U1 and U2 small nuclear ribonucleoproteins (snRNPs) bound to the precursor messenger RNA 5' splice site (5'SS) and branch point sequence, associates with the U4/U6.U5 tri-snRNP to form the fully assembled precatalytic pre-B spliceosome. Here, we report cryo-electron microscopy structures of the human pre-B complex captured before U1 snRNP dissociation at 3.3-angstrom core resolution and the human tri-snRNP at 2.9-angstrom resolution. U1 snRNP inserts the 5'SS-U1 snRNA helix between the two RecA domains of the Prp28 DEAD-box helicase. Adenosine 5'-triphosphate-dependent closure of the Prp28 RecA domains releases the 5'SS to pair with the nearby U6 ACAGAGA-box sequence presented as a mobile loop. The structures suggest that formation of the 5'SS-ACAGAGA helix triggers remodeling of an intricate protein-RNA network to induce Brr2 helicase relocation to its loading sequence in U4 snRNA, enabling Brr2 to unwind the U4/U6 snRNA duplex to allow U6 snRNA to form the catalytic center of the spliceosome. | |||||||||
履歴 |
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-構造の表示
ムービー |
ムービービューア |
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構造ビューア | 分子: MolmilJmol/JSmol |
-ダウンロードとリンク
-ダウンロード
PDBx/mmCIF形式 | 6qx9.cif.gz | 3.2 MB | 表示 | PDBx/mmCIF形式 |
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PDB形式 | pdb6qx9.ent.gz | 表示 | PDB形式 | |
PDBx/mmJSON形式 | 6qx9.json.gz | ツリー表示 | PDBx/mmJSON形式 | |
その他 | その他のダウンロード |
-検証レポート
アーカイブディレクトリ | https://data.pdbj.org/pub/pdb/validation_reports/qx/6qx9 ftp://data.pdbj.org/pub/pdb/validation_reports/qx/6qx9 | HTTPS FTP |
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-関連構造データ
関連構造データ | 4665MC 4658C 4672C 4673C 4674C 4675C 4676C 4686C 4687C 4688C 4689C 4690C 6qw6C M: このデータのモデリングに利用したマップデータ C: 同じ文献を引用 (文献) |
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類似構造データ | |
電子顕微鏡画像生データ | EMPIAR-10307 (タイトル: Human pre-B spliceosome and U4/U6.U5 tri-snRNP Data size: 2.3 TB Data #1: Dataset 1 of human pre-B spliceosome; motion-corrected micrographs [micrographs - single frame] Data #2: Dataset 2 of human pre-B spliceosome; motion-corrected micrographs [micrographs - single frame] Data #3: Dataset 3 of human pre-B spliceosome; motion-corrected micrographs [micrographs - single frame] Data #4: Dataset 4 of human pre-B spliceosome; motion-corrected micrographs [micrographs - single frame] Data #5: Dataset 5 of human pre-B spliceosome; motion-corrected micrographs [micrographs - single frame] Data #6: Selected U4/U6.U5 tri-snRNP particles after Bayesian polishing [picked particles - single frame - processed] Data #7: Crude shifted preB particles [picked particles - single frame - unprocessed]) |
-リンク
-集合体
登録構造単位 |
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1 |
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-要素
+RNA鎖 , 6種, 6分子 16I524
+U5 small nuclear ribonucleoprotein ... , 2種, 2分子 5O5B
+Splicing factor 3B subunit ... , 5種, 5分子 B4B2B5B3B1
+Small nuclear ribonucleoprotein ... , 6種, 24分子 135323435e4e1e2e411151211f2f5f4f224252121g2g5g4g
+U4/U6 small nuclear ribonucleoprotein ... , 3種, 3分子 4B4C4A
+U1 small nuclear ribonucleoprotein ... , 3種, 3分子 1K1A1C
+タンパク質 , 11種, 14分子 RX5C5X1b2b5b4b5J4D5A5DBPK
+U6 snRNA-associated Sm-like protein ... , 7種, 7分子 66676263686465
+U2 small nuclear ribonucleoprotein ... , 2種, 2分子 2B2A
+Splicing factor 3A subunit ... , 3種, 3分子 A2A3A1
+U4/U6.U5 tri-snRNP-associated protein ... , 2種, 2分子 SU
+非ポリマー , 4種, 9分子
-実験情報
-実験
実験 | 手法: 電子顕微鏡法 |
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EM実験 | 試料の集合状態: PARTICLE / 3次元再構成法: 単粒子再構成法 |
-試料調製
構成要素 |
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分子量 | 値: 3.0 MDa / 実験値: NO | ||||||||||||||||||||||||||||||
由来(天然) |
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由来(組換発現) |
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緩衝液 | pH: 7.9 | ||||||||||||||||||||||||||||||
緩衝液成分 |
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試料 | 濃度: 1.4 mg/ml / 包埋: NO / シャドウイング: NO / 染色: NO / 凍結: YES | ||||||||||||||||||||||||||||||
試料支持 | グリッドの材料: COPPER / グリッドのサイズ: 400 divisions/in. / グリッドのタイプ: Quantifoil R2/2 | ||||||||||||||||||||||||||||||
急速凍結 | 装置: FEI VITROBOT MARK III / 凍結剤: ETHANE / 湿度: 100 % / 凍結前の試料温度: 277 K |
-電子顕微鏡撮影
実験機器 | モデル: Titan Krios / 画像提供: FEI Company |
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顕微鏡 | モデル: FEI TITAN KRIOS |
電子銃 | 電子線源: FIELD EMISSION GUN / 加速電圧: 300 kV / 照射モード: FLOOD BEAM |
電子レンズ | モード: BRIGHT FIELDBright-field microscopy / アライメント法: COMA FREE |
試料ホルダ | 凍結剤: NITROGEN 試料ホルダーモデル: FEI TITAN KRIOS AUTOGRID HOLDER |
撮影 | 平均露光時間: 6 sec. / 電子線照射量: 50 e/Å2 / 検出モード: COUNTING フィルム・検出器のモデル: GATAN K2 SUMMIT (4k x 4k) |
電子光学装置 | エネルギーフィルター名称: GIF Bioquantum / エネルギーフィルタースリット幅: 20 eV |
画像スキャン | 動画フレーム数/画像: 40 / 利用したフレーム数/画像: 1-40 |
-解析
EMソフトウェア |
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CTF補正 | タイプ: PHASE FLIPPING AND AMPLITUDE CORRECTION | ||||||||||||||||||||||||||||||||
粒子像の選択 | 選択した粒子像数: 1130953 | ||||||||||||||||||||||||||||||||
対称性 | 点対称性: C1 (非対称) | ||||||||||||||||||||||||||||||||
3次元再構成 | 解像度: 3.28 Å / 解像度の算出法: FSC 0.143 CUT-OFF / 粒子像の数: 86146 / アルゴリズム: FOURIER SPACE / 対称性のタイプ: POINT |