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基本情報
登録情報 | データベース: PDB / ID: 8pk7 | |||||||||
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タイトル | Helical reconstruction of CHIKV nsP3 helical scaffolds | |||||||||
![]() | Non-structural protein 3 | |||||||||
![]() | VIRAL PROTEIN / Helical scaffold / Replication complex / Alpha granules / Viral factories | |||||||||
機能・相同性 | ![]() ADP-ribose 1''-phosphate phosphatase / host cell filopodium / mRNA methyltransferase activity / mRNA 5'-triphosphate monophosphatase activity / mRNA 5'-phosphatase / polynucleotide adenylyltransferase / polynucleotide 5'-phosphatase activity / poly(A) RNA polymerase activity / mRNA modification / regulation of cytoskeleton organization ...ADP-ribose 1''-phosphate phosphatase / host cell filopodium / mRNA methyltransferase activity / mRNA 5'-triphosphate monophosphatase activity / mRNA 5'-phosphatase / polynucleotide adenylyltransferase / polynucleotide 5'-phosphatase activity / poly(A) RNA polymerase activity / mRNA modification / regulation of cytoskeleton organization / symbiont-mediated suppression of host mRNA transcription via inhibition of RNA polymerase II activity / 7-methylguanosine mRNA capping / symbiont-mediated suppression of host JAK-STAT cascade via inhibition of STAT1 activity / cysteine-type peptidase activity / 転移酵素; 一炭素原子の基を移すもの; メチル基を移すもの / host cell cytoplasmic vesicle membrane / 転移酵素; リンを含む基を移すもの; 核酸を移すもの / nucleoside-triphosphate phosphatase / methylation / 加水分解酵素; プロテアーゼ; ペプチド結合加水分解酵素; システインプロテアーゼ / RNA helicase activity / RNA helicase / symbiont-mediated suppression of host innate immune response / symbiont-mediated suppression of host type I interferon-mediated signaling pathway / symbiont-mediated suppression of host gene expression / RNA-directed RNA polymerase / viral RNA genome replication / RNA-directed RNA polymerase activity / DNA-templated transcription / GTP binding / host cell nucleus / host cell plasma membrane / ATP hydrolysis activity / proteolysis / RNA binding / ATP binding / metal ion binding / membrane 類似検索 - 分子機能 | |||||||||
生物種 | ![]() | |||||||||
手法 | 電子顕微鏡法 / らせん対称体再構成法 / クライオ電子顕微鏡法 / 解像度: 2.52 Å | |||||||||
![]() | Reguera, J. / Hons, M. / Zimberger, C. / Ptchelkine, D. / Jones, R. / Desfosses, A. | |||||||||
資金援助 | ![]()
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![]() | ![]() タイトル: Alphavirus nsP3 organizes into tubular scaffolds essential for infection and the cytoplasmic granule architecture. 著者: Vasiliya Kril / Michael Hons / Celine Amadori / Claire Zimberger / Laurine Couture / Yara Bouery / Julien Burlaud-Gaillard / Andrei Karpov / Denis Ptchelkine / Alexandra L Thienel / Beate M ...著者: Vasiliya Kril / Michael Hons / Celine Amadori / Claire Zimberger / Laurine Couture / Yara Bouery / Julien Burlaud-Gaillard / Andrei Karpov / Denis Ptchelkine / Alexandra L Thienel / Beate M Kümmerer / Ambroise Desfosses / Rhian Jones / Philippe Roingeard / Laurent Meertens / Ali Amara / Juan Reguera / ![]() ![]() 要旨: Alphaviruses, such as chikungunya virus (CHIKV), are mosquito-borne viruses that represent a significant threat to human health due to the current context of global warming. Efficient alphavirus ...Alphaviruses, such as chikungunya virus (CHIKV), are mosquito-borne viruses that represent a significant threat to human health due to the current context of global warming. Efficient alphavirus infection relies on the activity of the non-structural protein 3 (nsP3), a puzzling multifunctional molecule whose role in infection remains largely unknown. NsP3 is a component of the plasma membrane-bound viral RNA replication complex (vRC) essential for RNA amplification and is also found in large cytoplasmic aggregates of unknown function Here, we report the cryo-electron microscopy (cryo-EM) structure of the CHIKV nsP3 at 2.35 Å resolution. We show that nsP3 assembles into tubular structures made by a helical arrangement of its alphavirus unique domain (AUD). The nsP3 helical scaffolds are consistent with crown structures found on tomographic reconstructions of the mature viral RCs. In addition, nsP3 helices assemble into cytoplasmic granules organized in a network of tubular structures that contain viral genomic RNA and capsid as well as host factors required for productive infection. Structure-guided mutagenesis identified residues that prevent or disturb nsP3 assemblies, resulting in impaired viral replication or transcription. Altogether, our results reveal an unexpected nsP3-dependent molecular organization essential for different phases of alphavirus infection. | |||||||||
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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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PDBx/mmCIF形式 | ![]() | 48.7 KB | 表示 | ![]() |
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PDB形式 | ![]() | 29 KB | 表示 | ![]() |
PDBx/mmJSON形式 | ![]() | ツリー表示 | ![]() | |
その他 | ![]() |
-検証レポート
文書・要旨 | ![]() | 1.2 MB | 表示 | ![]() |
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文書・詳細版 | ![]() | 1.2 MB | 表示 | |
XML形式データ | ![]() | 28.4 KB | 表示 | |
CIF形式データ | ![]() | 37.9 KB | 表示 | |
アーカイブディレクトリ | ![]() ![]() | HTTPS FTP |
-関連構造データ
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リンク
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集合体
登録構造単位 | ![]()
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1 | ![]()
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要素
#1: タンパク質 | 分子量: 57418.207 Da / 分子数: 1 / 由来タイプ: 組換発現 由来: (組換発現) ![]() 発現宿主: ![]() 参照: UniProt: Q8JUX6, ADP-ribose 1''-phosphate phosphatase |
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#2: 化合物 | ChemComp-ZN / |
研究の焦点であるリガンドがあるか | Y |
Has protein modification | 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: HELICAL ARRAY / 3次元再構成法: らせん対称体再構成法 |
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試料調製
構成要素 | 名称: Helical scaffold assembly of CHIKV nsP3 mediated by its Unique alphavirus domain. タイプ: COMPLEX / Entity ID: #1 / 由来: RECOMBINANT |
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分子量 | 値: 220.25 kDa/nm / 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 | 包埋: NO / シャドウイング: NO / 染色: NO / 凍結: YES / 詳細: his sample was heterogeneous in lenght |
急速凍結 | 凍結剤: ETHANE-PROPANE |
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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: FEI TITAN 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): 800 nm |
撮影 | 電子線照射量: 18.1 e/Å2 フィルム・検出器のモデル: GATAN K3 BIOQUANTUM (6k x 4k) |
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解析
EMソフトウェア |
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CTF補正 | タイプ: PHASE FLIPPING AND AMPLITUDE CORRECTION | |||||||||
らせん対称 | 回転角度/サブユニット: -164.175 ° / 軸方向距離/サブユニット: 2.782 Å / らせん対称軸の対称性: C1 | |||||||||
3次元再構成 | 解像度: 2.52 Å / 解像度の算出法: FSC 0.143 CUT-OFF / 粒子像の数: 807973 詳細: The assembly was calculated from the helical reconstruction of nsP3 from PDB 8PHZ subunit A. This was refined on a local refinement map at 2.35 A resolution 対称性のタイプ: HELICAL | |||||||||
原子モデル構築 | プロトコル: AB INITIO MODEL | |||||||||
原子モデル構築 | PDB-ID: 8PHZ PDB chain-ID: A / Accession code: 8PHZ / Chain residue range: 174-600 / Pdb chain residue range: 174-600 / Source name: PDB / タイプ: experimental model |