+データを開く
-基本情報
登録情報 | データベース: PDB / ID: 7ad8 | ||||||
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タイトル | Core TFIIH-XPA-DNA complex with modelled p62 subunit | ||||||
要素 |
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キーワード | DNA BINDING PROTEIN / DNA repair / transcription factor | ||||||
機能・相同性 | 機能・相同性情報 nucleotide-excision repair factor 1 complex / nucleotide-excision repair involved in interstrand cross-link repair / nucleotide-excision repair, DNA damage recognition / MMXD complex / core TFIIH complex portion of holo TFIIH complex / Cytosolic iron-sulfur cluster assembly / nucleotide-excision repair, DNA duplex unwinding / central nervous system myelin formation / response to auditory stimulus / positive regulation of mitotic recombination ...nucleotide-excision repair factor 1 complex / nucleotide-excision repair involved in interstrand cross-link repair / nucleotide-excision repair, DNA damage recognition / MMXD complex / core TFIIH complex portion of holo TFIIH complex / Cytosolic iron-sulfur cluster assembly / nucleotide-excision repair, DNA duplex unwinding / central nervous system myelin formation / response to auditory stimulus / positive regulation of mitotic recombination / hair follicle maturation / hair cell differentiation / nucleotide-excision repair factor 3 complex / nucleotide-excision repair, preincision complex assembly / CAK-ERCC2 complex / UV protection / embryonic cleavage / DNA 5'-3' helicase / G protein-coupled receptor internalization / UV-damage excision repair / transcription factor TFIIH holo complex / transcription factor TFIIH core complex / DNA 3'-5' helicase / RNA Polymerase I Transcription Termination / transcription preinitiation complex / nuclear thyroid hormone receptor binding / regulation of mitotic cell cycle phase transition / hematopoietic stem cell proliferation / 3'-5' DNA helicase activity / regulation of cyclin-dependent protein serine/threonine kinase activity / RNA Pol II CTD phosphorylation and interaction with CE during HIV infection / RNA Pol II CTD phosphorylation and interaction with CE / intercellular bridge / transcription factor TFIID complex / Formation of the Early Elongation Complex / Formation of the HIV-1 Early Elongation Complex / RNA polymerase II general transcription initiation factor activity / bone mineralization / spinal cord development / mRNA Capping / HIV Transcription Initiation / RNA Polymerase II HIV Promoter Escape / Transcription of the HIV genome / 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 / erythrocyte maturation / RNA Polymerase I Transcription Initiation / transcription by RNA polymerase I / ATPase activator activity / DNA topological change / intrinsic apoptotic signaling pathway by p53 class mediator / protein localization to nucleus / hematopoietic stem cell differentiation / Tat-mediated elongation of the HIV-1 transcript / embryonic organ development / transcription elongation by RNA polymerase I / Formation of HIV-1 elongation complex containing HIV-1 Tat / transcription-coupled nucleotide-excision repair / Formation of HIV elongation complex in the absence of HIV Tat / RNA Polymerase II Transcription Elongation / Formation of RNA Pol II elongation complex / response to UV / RNA Polymerase II Pre-transcription Events / DNA helicase activity / hormone-mediated signaling pathway / post-embryonic development / extracellular matrix organization / insulin-like growth factor receptor signaling pathway / maturation of SSU-rRNA from tricistronic rRNA transcript (SSU-rRNA, 5.8S rRNA, LSU-rRNA) / determination of adult lifespan / isomerase activity / chromosome segregation / promoter-specific chromatin binding / TP53 Regulates Transcription of DNA Repair Genes / nucleotide-excision repair / transcription initiation at RNA polymerase II promoter / RNA Polymerase I Promoter Escape / transcription elongation by RNA polymerase II / NoRC negatively regulates rRNA expression / base-excision repair / multicellular organism growth / Transcription-Coupled Nucleotide Excision Repair (TC-NER) / Formation of TC-NER Pre-Incision Complex / cellular response to gamma radiation / Dual Incision in GG-NER / Formation of Incision Complex in GG-NER / spindle / Dual incision in TC-NER / Gap-filling DNA repair synthesis and ligation in TC-NER / sequence-specific double-stranded DNA binding / protein localization / protein-macromolecule adaptor activity / 4 iron, 4 sulfur cluster binding / 5'-3' DNA helicase activity / double-stranded DNA binding / in utero embryonic development / response to oxidative stress / transcription by RNA polymerase II 類似検索 - 分子機能 | ||||||
生物種 | Homo sapiens (ヒト) | ||||||
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: 3.5 Å | ||||||
データ登録者 | Koelmel, W. / Kuper, J. / Kisker, C. | ||||||
資金援助 | ドイツ, 1件
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引用 | ジャーナル: Nucleic Acids Res / 年: 2020 タイトル: The TFIIH subunits p44/p62 act as a damage sensor during nucleotide excision repair. 著者: Jamie T Barnett / Jochen Kuper / Wolfgang Koelmel / Caroline Kisker / Neil M Kad / 要旨: Nucleotide excision repair (NER) in eukaryotes is orchestrated by the core form of the general transcription factor TFIIH, containing the helicases XPB, XPD and five 'structural' subunits, p62, p44, ...Nucleotide excision repair (NER) in eukaryotes is orchestrated by the core form of the general transcription factor TFIIH, containing the helicases XPB, XPD and five 'structural' subunits, p62, p44, p34, p52 and p8. Recent cryo-EM structures show that p62 makes extensive contacts with p44 and in part occupies XPD's DNA binding site. While p44 is known to regulate the helicase activity of XPD during NER, p62 is thought to be purely structural. Here, using helicase and adenosine triphosphatase assays we show that a complex containing p44 and p62 enhances XPD's affinity for dsDNA 3-fold over p44 alone. Remarkably, the relative affinity is further increased to 60-fold by dsDNA damage. Direct binding studies show this preference derives from p44/p62's high affinity (20 nM) for damaged ssDNA. Single molecule imaging of p44/p62 complexes without XPD reveals they bind to and randomly diffuse on DNA, however, in the presence of UV-induced DNA lesions these complexes stall. Combined with the analysis of a recent cryo-EM structure, we suggest that p44/p62 acts as a novel DNA-binding entity that enhances damage recognition in TFIIH. This revises our understanding of TFIIH and prompts investigation into the core subunits for an active role during DNA repair and/or transcription. | ||||||
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 | 7ad8.cif.gz | 505.5 KB | 表示 | PDBx/mmCIF形式 |
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PDB形式 | pdb7ad8.ent.gz | 390.9 KB | 表示 | PDB形式 |
PDBx/mmJSON形式 | 7ad8.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
文書・要旨 | 7ad8_validation.pdf.gz | 808.6 KB | 表示 | wwPDB検証レポート |
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文書・詳細版 | 7ad8_full_validation.pdf.gz | 823.2 KB | 表示 | |
XML形式データ | 7ad8_validation.xml.gz | 67.1 KB | 表示 | |
CIF形式データ | 7ad8_validation.cif.gz | 104.4 KB | 表示 | |
アーカイブディレクトリ | https://data.pdbj.org/pub/pdb/validation_reports/ad/7ad8 ftp://data.pdbj.org/pub/pdb/validation_reports/ad/7ad8 | HTTPS FTP |
-関連構造データ
関連構造データ | 4970M M: このデータのモデリングに利用したマップデータ |
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類似構造データ |
-リンク
-集合体
登録構造単位 |
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1 |
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-要素
-DNA鎖 , 2種, 2分子 JK
#1: DNA鎖 | 分子量: 15036.663 Da / 分子数: 1 / 由来タイプ: 合成 / 由来: (合成) Homo sapiens (ヒト) |
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#2: DNA鎖 | 分子量: 15075.698 Da / 分子数: 1 / 由来タイプ: 合成 / 由来: (合成) Homo sapiens (ヒト) |
-タンパク質 , 3種, 3分子 ABG
#3: タンパク質 | 分子量: 89404.734 Da / 分子数: 1 / 由来タイプ: 組換発現 / 由来: (組換発現) Homo sapiens (ヒト) / 遺伝子: ERCC3, XPB, XPBC / 発現宿主: Trichoplusia ni (イラクサキンウワバ) / 参照: UniProt: P19447, DNA helicase |
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#5: タンパク質 | 分子量: 87021.078 Da / 分子数: 1 / 由来タイプ: 組換発現 / 由来: (組換発現) Homo sapiens (ヒト) / 遺伝子: ERCC2, XPD, XPDC / 発現宿主: Trichoplusia ni (イラクサキンウワバ) / 参照: UniProt: P18074, DNA helicase |
#9: タンパク質 | 分子量: 31422.053 Da / 分子数: 1 / 由来タイプ: 組換発現 / 由来: (組換発現) Homo sapiens (ヒト) / 遺伝子: XPA, XPAC / 発現宿主: Trichoplusia ni (イラクサキンウワバ) / 参照: UniProt: P23025 |
-General transcription factor IIH subunit ... , 5種, 5分子 FEDCI
#4: タンパク質 | 分子量: 8060.362 Da / 分子数: 1 / 由来タイプ: 組換発現 / 由来: (組換発現) Homo sapiens (ヒト) / 遺伝子: GTF2H5, C6orf175, TTDA / 発現宿主: Trichoplusia ni (イラクサキンウワバ) / 参照: UniProt: Q6ZYL4 |
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#6: タンパク質 | 分子量: 34416.008 Da / 分子数: 1 / 由来タイプ: 組換発現 / 由来: (組換発現) Homo sapiens (ヒト) / 遺伝子: GTF2H3 / 発現宿主: Trichoplusia ni (イラクサキンウワバ) / 参照: UniProt: Q13889 |
#7: タンパク質 | 分子量: 44481.996 Da / 分子数: 1 / 由来タイプ: 組換発現 / 由来: (組換発現) Homo sapiens (ヒト) / 遺伝子: GTF2H2, BTF2P44 / 発現宿主: Trichoplusia ni (イラクサキンウワバ) / 参照: UniProt: Q13888 |
#8: タンパク質 | 分子量: 52245.156 Da / 分子数: 1 / 由来タイプ: 組換発現 / 由来: (組換発現) Homo sapiens (ヒト) / 遺伝子: GTF2H4 / 発現宿主: Trichoplusia ni (イラクサキンウワバ) / 参照: UniProt: Q92759 |
#10: タンパク質 | 分子量: 62116.492 Da / 分子数: 1 / 由来タイプ: 組換発現 / 由来: (組換発現) Homo sapiens (ヒト) / 遺伝子: GTF2H1, BTF2 / 発現宿主: Trichoplusia ni (イラクサキンウワバ) / 参照: UniProt: P32780 |
-非ポリマー , 2種, 7分子
#11: 化合物 | ChemComp-SF4 / |
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#12: 化合物 | ChemComp-ZN / |
-詳細
研究の焦点であるリガンドがあるか | N |
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-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 | 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: 7.5 | ||||||||||||||||||||||||
試料 | 包埋: NO / シャドウイング: NO / 染色: NO / 凍結: YES | ||||||||||||||||||||||||
急速凍結 | 凍結剤: ETHANE |
-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 FIELD |
撮影 | 電子線照射量: 41 e/Å2 フィルム・検出器のモデル: GATAN K2 SUMMIT (4k x 4k) |
-解析
CTF補正 | タイプ: PHASE FLIPPING AND AMPLITUDE CORRECTION |
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3次元再構成 | 解像度: 3.5 Å / 解像度の算出法: FSC 0.143 CUT-OFF / 粒子像の数: 227776 / 対称性のタイプ: POINT |