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基本情報
登録情報 | データベース: PDB / ID: 8soj | |||||||||
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タイトル | Cryo-EM structure of human CST bound to POT1(ESDL)/TPP1 in the absence of telomeric ssDNA | |||||||||
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![]() | DNA BINDING PROTEIN / telomere / shelterin / cst / complex | |||||||||
機能・相同性 | ![]() positive regulation of single-stranded telomeric DNA binding / positive regulation of DNA strand elongation / regulation of DNA helicase activity / positive regulation of DNA helicase activity / G-rich single-stranded DNA binding / telomere assembly / CST complex / segmentation / urogenital system development / 8-hydroxy-2'-deoxyguanosine DNA binding ...positive regulation of single-stranded telomeric DNA binding / positive regulation of DNA strand elongation / regulation of DNA helicase activity / positive regulation of DNA helicase activity / G-rich single-stranded DNA binding / telomere assembly / CST complex / segmentation / urogenital system development / 8-hydroxy-2'-deoxyguanosine DNA binding / telomeric D-loop binding / regulation of double-strand break repair via nonhomologous end joining / positive regulation of helicase activity / protection from non-homologous end joining at telomere / telomerase inhibitor activity / DEAD/H-box RNA helicase binding / establishment of protein localization to telomere / telomere maintenance via telomere lengthening / telomeric D-loop disassembly / shelterin complex / Telomere C-strand synthesis initiation / regulation of telomere maintenance via telomerase / Telomere C-strand (Lagging Strand) Synthesis / : / positive regulation of telomere maintenance / single-stranded telomeric DNA binding / nuclear telomere cap complex / Processive synthesis on the C-strand of the telomere / Polymerase switching on the C-strand of the telomere / Removal of the Flap Intermediate from the C-strand / G-rich strand telomeric DNA binding / telomere capping / telomerase holoenzyme complex / bone marrow development / embryonic limb morphogenesis / intermediate filament cytoskeleton / protein localization to chromosome, telomeric region / DNA duplex unwinding / hematopoietic stem cell proliferation / telomeric DNA binding / : / negative regulation of telomere maintenance via telomerase / symbiont-mediated suppression of host cytoplasmic pattern recognition receptor signaling pathway via inhibition of MDA-5 activity / Telomere Extension By Telomerase / carbohydrate transmembrane transporter activity / telomere maintenance via telomerase / maltose binding / replicative senescence / maltose transport / maltodextrin transmembrane transport / DNA polymerase binding / ATP-binding cassette (ABC) transporter complex, substrate-binding subunit-containing / Packaging Of Telomere Ends / Recognition and association of DNA glycosylase with site containing an affected purine / Cleavage of the damaged purine / spleen development / positive regulation of telomere maintenance via telomerase / Recognition and association of DNA glycosylase with site containing an affected pyrimidine / Cleavage of the damaged pyrimidine / regulation of G2/M transition of mitotic cell cycle / Inhibition of DNA recombination at telomere / Meiotic synapsis / picornain 2A / symbiont-mediated suppression of host mRNA export from nucleus / telomere maintenance / symbiont genome entry into host cell via pore formation in plasma membrane / bioluminescence / picornain 3C / thymus development / T=pseudo3 icosahedral viral capsid / positive regulation of DNA replication / generation of precursor metabolites and energy / skeletal system development / host cell cytoplasmic vesicle membrane / intracellular protein transport / cytoplasmic vesicle membrane / multicellular organism growth / endocytosis involved in viral entry into host cell / DNA Damage/Telomere Stress Induced Senescence / fibrillar center / symbiont-mediated suppression of host gene expression / positive regulation of fibroblast proliferation / nucleoside-triphosphate phosphatase / protein complex oligomerization / monoatomic ion channel activity / outer membrane-bounded periplasmic space / single-stranded DNA binding / chromosome, telomeric region / RNA helicase activity / nuclear body / induction by virus of host autophagy / RNA-directed RNA polymerase / viral RNA genome replication / cysteine-type endopeptidase activity / intracellular membrane-bounded organelle / RNA-dependent RNA polymerase activity / virus-mediated perturbation of host defense response / DNA-templated transcription / DNA damage response / host cell nucleus 類似検索 - 分子機能 | |||||||||
生物種 | ![]() ![]() ![]() ![]() ![]() | |||||||||
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: 3.8 Å | |||||||||
![]() | Cai, S.W. | |||||||||
資金援助 | ![]()
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![]() | ![]() タイトル: POT1 recruits and regulates CST-Polα/Primase at human telomeres. 著者: Sarah W Cai / Hiroyuki Takai / Thomas Walz / Titia de Lange / ![]() 要旨: Telomere maintenance requires extension of the G-rich telomeric repeat strand by telomerase and fill-in synthesis of the C-rich strand by Polα/Primase. Telomeric Polα/Primase is bound to Ctc1-Stn1- ...Telomere maintenance requires extension of the G-rich telomeric repeat strand by telomerase and fill-in synthesis of the C-rich strand by Polα/Primase. Telomeric Polα/Primase is bound to Ctc1-Stn1-Ten1 (CST), a single-stranded DNA-binding complex. Like mutations in telomerase, mutations affecting CST-Polα/Primase result in pathological telomere shortening and cause a telomere biology disorder, Coats plus (CP). We determined cryogenic electron microscopy structures of human CST bound to the shelterin heterodimer POT1/TPP1 that reveal how CST is recruited to telomeres by POT1. Phosphorylation of POT1 is required for CST recruitment, and the complex is formed through conserved interactions involving several residues mutated in CP. Our structural and biochemical data suggest that phosphorylated POT1 holds CST-Polα/Primase in an inactive auto-inhibited state until telomerase has extended the telomere ends. We propose that dephosphorylation of POT1 releases CST-Polα/Primase into an active state that completes telomere replication through fill-in synthesis. | |||||||||
履歴 |
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構造の表示
構造ビューア | 分子: ![]() ![]() |
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ダウンロードとリンク
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ダウンロード
PDBx/mmCIF形式 | ![]() | 764.8 KB | 表示 | ![]() |
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PDB形式 | ![]() | 604.7 KB | 表示 | ![]() |
PDBx/mmJSON形式 | ![]() | ツリー表示 | ![]() | |
その他 | ![]() |
-検証レポート
アーカイブディレクトリ | ![]() ![]() | HTTPS FTP |
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-関連構造データ
関連構造データ | ![]() 40659MC ![]() 8sokC 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 ( |
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類似構造データ | 類似検索 - 機能・相同性 ![]() |
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リンク
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集合体
登録構造単位 | ![]()
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要素
#1: タンパク質 | 分子量: 178300.609 Da / 分子数: 1 / 由来タイプ: 組換発現 / 詳細: His6-MBP-3C tagged human Ctc1 由来: (組換発現) ![]() ![]() ![]() 遺伝子: malE, Z5632, ECs5017, CTC1, C17orf68 / 発現宿主: ![]() | ||
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#2: タンパク質 | 分子量: 115627.211 Da / 分子数: 1 / 由来タイプ: 組換発現 由来: (組換発現) ![]() ![]() ![]() 遺伝子: ACD, PIP1, PTOP, TINT1, TPP1, STN1, OBFC1 / 発現宿主: ![]() 参照: UniProt: B6F2F5, UniProt: Q96AP0, UniProt: Q9H668, picornain 2A, nucleoside-triphosphate phosphatase, picornain 3C, RNA-directed RNA polymerase | ||
#3: タンパク質 | 分子量: 13872.013 Da / 分子数: 1 / 由来タイプ: 組換発現 / 由来: (組換発現) ![]() ![]() | ||
#4: タンパク質 | 分子量: 72938.219 Da / 分子数: 1 / 由来タイプ: 組換発現 詳細: mPOT1b residues ESDL 323-326 are inserted into the human POT1 sequence between S320 and V321 由来: (組換発現) ![]() ![]() | ||
#5: 化合物 | 研究の焦点であるリガンドがあるか | N | |
-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 |
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試料調製
構成要素 | 名称: Human CST-POT1(ESDL)/TPP1 complex / タイプ: COMPLEX / 詳細: CST-POT1(ESDL)/TPP1 complex in the absence of DNA / Entity ID: #1-#4 / 由来: RECOMBINANT |
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分子量 | 値: 0.38 MDa / 実験値: NO |
由来(天然) | 生物種: ![]() |
由来(組換発現) | 生物種: ![]() |
緩衝液 | pH: 7.5 |
試料 | 濃度: 0.05 mg/ml / 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 |
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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: TFS KRIOS |
電子銃 | 電子線源: ![]() |
電子レンズ | モード: BRIGHT FIELD / 最大 デフォーカス(公称値): 2500 nm / 最小 デフォーカス(公称値): 1000 nm |
撮影 | 電子線照射量: 50.3 e/Å2 / フィルム・検出器のモデル: GATAN K3 (6k x 4k) |
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解析
ソフトウェア |
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CTF補正 | タイプ: PHASE FLIPPING AND AMPLITUDE CORRECTION | |||||||||
3次元再構成 | 解像度: 3.8 Å / 解像度の算出法: FSC 0.143 CUT-OFF / 粒子像の数: 132356 / 対称性のタイプ: POINT | |||||||||
精密化 | 交差検証法: NONE |