- EMDB-48622: Structure of a native Drosophila melanogaster Pol II Elongation C... -
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基本情報
登録情報
データベース: EMDB / ID: EMD-48622
タイトル
Structure of a native Drosophila melanogaster Pol II Elongation Complex with a well-defined Rpb4/Rpb7 stalk
マップデータ
Refined, unsharpened Coulomb potential density map. Particles extracted in 440x440 Fourier cropped to 360x360.
試料
複合体: Native purified Pol II elongation complex from Drosophila melanogaster with a well-defined Rpb4/Rpb7 stalk
タンパク質・ペプチド: x 12種
RNA: x 1種
DNA: x 2種
リガンド: x 1種
キーワード
polymerase / Pol II / transcription / mRNA
機能・相同性
機能・相同性情報
Formation of the Early Elongation Complex / mRNA Capping / mRNA Splicing - Minor Pathway / RNA Polymerase I Transcription Initiation / RNA Polymerase III Transcription Initiation From Type 1 Promoter / RNA Polymerase III Transcription Initiation From Type 2 Promoter / RNA Pol II CTD phosphorylation and interaction with CE / Formation of RNA Pol II elongation complex / TP53 Regulates Transcription of DNA Repair Genes / RNA Polymerase II Transcription Elongation ...Formation of the Early Elongation Complex / mRNA Capping / mRNA Splicing - Minor Pathway / RNA Polymerase I Transcription Initiation / RNA Polymerase III Transcription Initiation From Type 1 Promoter / RNA Polymerase III Transcription Initiation From Type 2 Promoter / RNA Pol II CTD phosphorylation and interaction with CE / Formation of RNA Pol II elongation complex / TP53 Regulates Transcription of DNA Repair Genes / RNA Polymerase II Transcription Elongation / RNA Polymerase II Promoter Escape / RNA Polymerase II Transcription Pre-Initiation And Promoter Opening / RNA Polymerase II Transcription Initiation / RNA Polymerase II Transcription Initiation And Promoter Clearance / polytene chromosome interband / RNA Polymerase II Pre-transcription Events / Gap-filling DNA repair synthesis and ligation in TC-NER / Formation of TC-NER Pre-Incision Complex / Processing of Capped Intron-Containing Pre-mRNA / Dual incision in TC-NER / polytene chromosome puff / RNA polymerase II transcribes snRNA genes / mRNA Splicing - Major Pathway / RNA Polymerase I Promoter Escape / Transcriptional regulation by small RNAs / Estrogen-dependent gene expression / polytene chromosome / positive regulation of nuclear-transcribed mRNA poly(A) tail shortening / maintenance of transcriptional fidelity during transcription elongation by RNA polymerase II / positive regulation of translational initiation / nuclear-transcribed mRNA catabolic process / RNA polymerase I complex / RNA polymerase III complex / transcription elongation by RNA polymerase I / RNA polymerase II, core complex / tRNA transcription by RNA polymerase III / transcription by RNA polymerase I / transcription-coupled nucleotide-excision repair / translation initiation factor binding / transcription initiation at RNA polymerase II promoter / P-body / mRNA transcription by RNA polymerase II / ribonucleoside binding / DNA-directed RNA polymerase / DNA-directed RNA polymerase activity / mitotic cell cycle / single-stranded DNA binding / cellular response to heat / transcription by RNA polymerase II / nucleic acid binding / protein dimerization activity / single-stranded RNA binding / nucleotide binding / nucleolus / DNA binding / zinc ion binding / metal ion binding / nucleus / cytoplasm 類似検索 - 分子機能
DNA-directed RNA polymerase II subunit RPB3 / DNA-directed RNA polymerase II subunit RPB1 / DNA-directed RNA polymerase II subunit RPB2 / DNA-directed RNA polymerase II subunit RPB9 / DNA-directed RNA polymerases I, II, and III subunit RPABC2 / DNA-directed RNA polymerases I, II, and III subunit RPABC4 / DNA-directed RNA polymerases I, II, and III subunit RPABC1 / DNA-directed RNA polymerases I, II, and III subunit RPABC5 / DNA-directed RNA polymerase II subunit Rpb4 / DNA-directed RNA polymerase II subunit RPB7 ...DNA-directed RNA polymerase II subunit RPB3 / DNA-directed RNA polymerase II subunit RPB1 / DNA-directed RNA polymerase II subunit RPB2 / DNA-directed RNA polymerase II subunit RPB9 / DNA-directed RNA polymerases I, II, and III subunit RPABC2 / DNA-directed RNA polymerases I, II, and III subunit RPABC4 / DNA-directed RNA polymerases I, II, and III subunit RPABC1 / DNA-directed RNA polymerases I, II, and III subunit RPABC5 / DNA-directed RNA polymerase II subunit Rpb4 / DNA-directed RNA polymerase II subunit RPB7 / DNA-directed RNA polymerase II subunit RPB11 / DNA-directed RNA polymerases I, II, and III subunit RPABC3 類似検索 - 構成要素
全体 : Native purified Pol II elongation complex from Drosophila melanog...
全体
名称: Native purified Pol II elongation complex from Drosophila melanogaster with a well-defined Rpb4/Rpb7 stalk
要素
複合体: Native purified Pol II elongation complex from Drosophila melanogaster with a well-defined Rpb4/Rpb7 stalk
タンパク質・ペプチド: DNA-directed RNA polymerase II subunit RPB1
タンパク質・ペプチド: DNA-directed RNA polymerase II subunit RPB2
タンパク質・ペプチド: DNA-directed RNA polymerase II subunit RPB3
タンパク質・ペプチド: DNA-directed RNA polymerase II subunit Rpb4
タンパク質・ペプチド: DNA-directed RNA polymerases I, II, and III subunit RPABC2
タンパク質・ペプチド: DNA-directed RNA polymerase II subunit RPB7
タンパク質・ペプチド: DNA-directed RNA polymerases I, II, and III subunit RPABC3
タンパク質・ペプチド: DNA-directed RNA polymerase II subunit RPB9
タンパク質・ペプチド: DNA-directed RNA polymerases I, II, and III subunit RPABC5
タンパク質・ペプチド: DNA-directed RNA polymerase II subunit RPB11
タンパク質・ペプチド: DNA-directed RNA polymerase II subunit RPB12
RNA: RNA
DNA: Template DNA
DNA: Non-template DNA
タンパク質・ペプチド: DNA-directed RNA polymerase II subunit RPB5
リガンド: ZINC ION
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超分子 #1: Native purified Pol II elongation complex from Drosophila melanog...
超分子
名称: Native purified Pol II elongation complex from Drosophila melanogaster with a well-defined Rpb4/Rpb7 stalk タイプ: complex / ID: 1 / 親要素: 0 / 含まれる分子: #1-#15 詳細: This entry represents a native Pol II elongation complex from Drosophila melanogaster with a well-defined Rpb4/Rpb7 stalk, obtained through stringent 3D classification and refinement.
由来(天然)
生物種: Drosophila melanogaster (キイロショウジョウバエ)
分子量
理論値: 930 kDa/nm
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分子 #1: DNA-directed RNA polymerase II subunit RPB1
凍結剤: ETHANE / チャンバー内湿度: 100 % / チャンバー内温度: 4 K / 装置: FEI VITROBOT MARK IV
詳細
double FLAG-tagged Pol II subunit Rpb1 was used for purification of Pol II complexes
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電子顕微鏡法
顕微鏡
TFS TALOS
撮影
フィルム・検出器のモデル: FEI FALCON IV (4k x 4k) デジタル化 - サイズ - 横: 4096 pixel / デジタル化 - サイズ - 縦: 4096 pixel / 撮影したグリッド数: 4 / 実像数: 31103 / 平均電子線量: 50.65 e/Å2 詳細: Although the data was collected, motion-corrected and CTF estimated using the pixel size of 0.944, all the subsequent data processing was performed using pixel size of 1.1538. This was ...詳細: Although the data was collected, motion-corrected and CTF estimated using the pixel size of 0.944, all the subsequent data processing was performed using pixel size of 1.1538. This was achieved by extracting particles in box size 440x440 and Fourier-cropping it to box size 360x360
選択した数: 2349308 詳細: Initially, nearly 15 million particles were picked from micrographs. However, the number of particles cited here (2,349,308) is the starting number of particles after initial cleanup and 3D ...詳細: Initially, nearly 15 million particles were picked from micrographs. However, the number of particles cited here (2,349,308) is the starting number of particles after initial cleanup and 3D classification of the data.
CTF補正
ソフトウェア - 名称: cryoSPARC (ver. 4.3.1) / 詳細: 'Patch CTF Estimation' in cryoSPARC / タイプ: PHASE FLIPPING AND AMPLITUDE CORRECTION
初期モデル
モデルのタイプ: INSILICO MODEL / In silico モデル: 'Ab initio' function in cryoSPARC
最終 再構成
使用したクラス数: 1 / 想定した対称性 - 点群: C1 (非対称) / アルゴリズム: BACK PROJECTION / 解像度のタイプ: BY AUTHOR / 解像度: 3.44 Å / 解像度の算出法: FSC 0.143 CUT-OFF / ソフトウェア - 名称: cryoSPARC (ver. 4.3.1) 詳細: The data was processed using cryoSPARC to 3.44 A from 70,116 particles 使用した粒子像数: 70116
初期 角度割当
タイプ: MAXIMUM LIKELIHOOD / ソフトウェア - 名称: cryoSPARC (ver. 4.3.1)
最終 角度割当
タイプ: MAXIMUM LIKELIHOOD / ソフトウェア - 名称: cryoSPARC (ver. 4.3.1) / 詳細: 'Non-Uniform Refinement' in cryoSPARC
chain_id: L, source_name: AlphaFold, initial_model_type: in silico model
詳細
The model was based on PDB: 6GML. First, 6GML was rigid-body fit in the cryo-EM Coulomb potential density map using UCSF ChimeraX. Next, AlphaFold2 models for individual Drosophila melanogaster Pol II subunits were aligned to their 6GML counterparts, and further optimized in the density using rigid-body fit. For the final model optimization, phenix.real_space_refine was used
精密化
プロトコル: AB INITIO MODEL / 温度因子: 50
得られたモデル
PDB-9mu7: Structure of a native Drosophila melanogaster Pol II Elongation Complex with a well-defined Rpb4/Rpb7 stalk