- EMDB-48626: Structure of a native Drosophila melanogaster Nucleosome Elongati... -
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基本情報
登録情報
データベース: EMDB / ID: EMD-48626
タイトル
Structure of a native Drosophila melanogaster Nucleosome Elongation Complex (Pol II EC-nucleosome). Composite map
マップデータ
Composite map of the locally refined Pol II and nucleosome from the Nucleosome Elongation Complex.
試料
複合体: Native purified Nucleosome Elongation Complex from Drosophila melanogaster
RNA: x 1種
DNA: x 2種
タンパク質・ペプチド: x 14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 / 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 / HDMs demethylate histones / PKMTs methylate histone lysines / Interleukin-7 signaling / Chromatin modifying enzymes / SUMOylation of chromatin organization proteins / Recruitment and ATM-mediated phosphorylation of repair and signaling proteins at DNA double strand breaks / mRNA Polyadenylation / RMTs methylate histone arginines / E3 ubiquitin ligases ubiquitinate target proteins / RCAF complex / Factors involved in megakaryocyte development and platelet production / SIRT1 negatively regulates rRNA expression / NoRC negatively regulates rRNA expression / Activated PKN1 stimulates transcription of AR (androgen receptor) regulated genes KLK2 and KLK3 / polytene chromosome band / PRC2 methylates histones and DNA / Ub-specific processing proteases / Negative Regulation of CDH1 Gene Transcription / MLL4 and MLL3 complexes regulate expression of PPARG target genes in adipogenesis and hepatic steatosis / Regulation of PD-L1(CD274) transcription / larval somatic muscle development / RNA Polymerase I Promoter Escape / RUNX1 regulates genes involved in megakaryocyte differentiation and platelet function / Senescence-Associated Secretory Phenotype (SASP) / Regulation of endogenous retroelements by KRAB-ZFP proteins / Transcriptional regulation by small RNAs / Estrogen-dependent gene expression / HATs acetylate histones / Assembly of the ORC complex at the origin of replication / Oxidative Stress Induced Senescence / polytene chromosome / nuclear chromosome / termination of RNA polymerase II transcription / transcription by RNA polymerase III / maintenance of transcriptional fidelity during transcription elongation by RNA polymerase II / nucleolar large rRNA transcription by RNA polymerase I / transcription initiation at RNA polymerase I promoter / termination of RNA polymerase III transcription / transcription initiation at RNA polymerase III promoter / RNA polymerase I complex / RNA polymerase III complex / RNA polymerase II, core complex / transcription elongation by RNA polymerase I / tRNA transcription by RNA polymerase III / transcription by RNA polymerase I / transcription-coupled nucleotide-excision repair / DNA-templated transcription elongation / transcription initiation at RNA polymerase II promoter / mRNA transcription by RNA polymerase II / transcription by RNA polymerase II / ribonucleoside binding / nucleosomal DNA binding / DNA-directed RNA polymerase / structural constituent of chromatin / DNA-directed RNA polymerase activity / nucleosome / mitotic cell cycle / nucleosome assembly / cellular response to heat / chromosome / heterochromatin formation / chromatin organization / nucleic acid binding / protein dimerization activity / protein heterodimerization activity / nucleolus / protein-containing complex binding / chromatin / 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 / Histone H2B / Histone H3 / DNA-directed RNA polymerase II subunit RPB1 / DNA-directed RNA polymerase II subunit RPB2 / DNA-directed RNA polymerase II subunit RPB9 / Histone H4 / Histone H2A / DNA-directed RNA polymerases I, II, and III subunit RPABC2 / DNA-directed RNA polymerases I, II, and III subunit RPABC4 ...DNA-directed RNA polymerase II subunit RPB3 / Histone H2B / Histone H3 / DNA-directed RNA polymerase II subunit RPB1 / DNA-directed RNA polymerase II subunit RPB2 / DNA-directed RNA polymerase II subunit RPB9 / Histone H4 / Histone H2A / 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 RPB11 / DNA-directed RNA polymerases I, II, and III subunit RPABC3 類似検索 - 構成要素
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: 4197404 詳細: Initially, nearly 15 million particles were picked from micrographs. However, the number of particles cited here (4,197,404) 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 (4,197,404) is the starting number of particles after initial cleanup and 3D classification of the data representing Pol II and nucleosome subsets, respectively.
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: 7.8 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 Nucleosome EC was refined to an overall resolution of 7.8 A. Then, local refinement was performed with an indvidual focus on the nucleosome and Pol II. This entry represents a composite ...詳細: The Nucleosome EC was refined to an overall resolution of 7.8 A. Then, local refinement was performed with an indvidual focus on the nucleosome and Pol II. This entry represents a composite map of the two locally refined entities. 使用した粒子像数: 36552
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
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原子モデル構築 1
詳細
The model was based on individual models from this study: octameric nucleosome and Pol II. The models were rigid-body fit into the composite structure, connected and fixed in Coot and Phenix.
精密化
プロトコル: AB INITIO MODEL / 温度因子: 50
得られたモデル
PDB-9mu9: Structure of a native Drosophila melanogaster Nucleosome Elongation Complex (Pol II EC-nucleosome). Composite map