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データを開く
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基本情報
登録情報 | データベース: PDB / ID: 6v86 | ||||||||||||||||||
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タイトル | Parainfluenza virus 5 L-P complex with an alternate conformation of the CD-MTase-CTD module | ||||||||||||||||||
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![]() | VIRAL PROTEIN / POLYMERASE / METHYLTRANSFERASE / POLY-RIBONUCLEOTIDYLTRANSFERASE | ||||||||||||||||||
機能・相同性 | ![]() NNS virus cap methyltransferase / GDP polyribonucleotidyltransferase / 加水分解酵素; 酸無水物に作用; リン含有酸無水物に作用 / virion component / mRNA 5'-cap (guanine-N7-)-methyltransferase activity / host cell cytoplasm / RNA-directed RNA polymerase / RNA-dependent RNA polymerase activity / GTPase activity / ATP binding 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: 4.63 Å | ||||||||||||||||||
![]() | Abdella, R. / He, Y. | ||||||||||||||||||
資金援助 | ![]()
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![]() | ![]() タイトル: Structure of a paramyxovirus polymerase complex reveals a unique methyltransferase-CTD conformation. 著者: Ryan Abdella / Megha Aggarwal / Takashi Okura / Robert A Lamb / Yuan He / ![]() 要旨: Paramyxoviruses are enveloped, nonsegmented, negative-strand RNA viruses that cause a wide spectrum of human and animal diseases. The viral genome, packaged by the nucleoprotein (N), serves as a ...Paramyxoviruses are enveloped, nonsegmented, negative-strand RNA viruses that cause a wide spectrum of human and animal diseases. The viral genome, packaged by the nucleoprotein (N), serves as a template for the polymerase complex, composed of the large protein (L) and the homo-tetrameric phosphoprotein (P). The ∼250-kDa L possesses all enzymatic activities necessary for its function but requires P in vivo. Structural information is available for individual P domains from different paramyxoviruses, but how P interacts with L and how that affects the activity of L is largely unknown due to the lack of high-resolution structures of this complex in this viral family. In this study we determined the structure of the L-P complex from parainfluenza virus 5 (PIV5) at 4.3-Å resolution using cryoelectron microscopy, as well as the oligomerization domain (OD) of P at 1.4-Å resolution using X-ray crystallography. P-OD associates with the RNA-dependent RNA polymerase domain of L and protrudes away from it, while the X domain of one chain of P is bound near the L nucleotide entry site. The methyltransferase (MTase) domain and the C-terminal domain (CTD) of L adopt a unique conformation, positioning the MTase active site immediately above the poly-ribonucleotidyltransferase domain and near the likely exit site for the product RNA 5' end. Our study reveals a potential mechanism that mononegavirus polymerases may employ to switch between transcription and genome replication. This knowledge will assist in the design and development of antivirals against paramyxoviruses. | ||||||||||||||||||
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構造の表示
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構造ビューア | 分子: ![]() ![]() |
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-検証レポート
文書・要旨 | ![]() | 789.8 KB | 表示 | ![]() |
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-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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要素
#1: タンパク質 | 分子量: 256195.672 Da / 分子数: 1 / 由来タイプ: 組換発現 由来: (組換発現) ![]() 株: W3 発現宿主: ![]() ![]() 参照: UniProt: Q88434, RNA-directed RNA polymerase, mRNA (guanine-N7)-methyltransferase, GDP polyribonucleotidyltransferase, EC: 2.1.1.296 | ||||
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#2: タンパク質 | 分子量: 42155.152 Da / 分子数: 5 / 由来タイプ: 組換発現 詳細: Chain F belongs to the same peptide as one of B, C, D or E, but the density is not well resolved enough to determine which chain it should associate with. 由来: (組換発現) ![]() 株: W3 / 遺伝子: P/V 発現宿主: ![]() ![]() 参照: UniProt: P11208 #3: 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
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分子量 |
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由来(天然) |
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緩衝液 | pH: 7.4 | ||||||||||||||||||||||||||||
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 |
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電子顕微鏡撮影
顕微鏡 | モデル: JEOL 3200FS |
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電子銃 | 電子線源: ![]() |
電子レンズ | モード: BRIGHT FIELD |
撮影 | 電子線照射量: 80.8 e/Å2 / 検出モード: SUPER-RESOLUTION フィルム・検出器のモデル: GATAN K2 SUMMIT (4k 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 | ||||||||||||||||||||||||||||
粒子像の選択 | 選択した粒子像数: 717008 | ||||||||||||||||||||||||||||
対称性 | 点対称性: C1 (非対称) | ||||||||||||||||||||||||||||
3次元再構成 | 解像度: 4.63 Å / 解像度の算出法: FSC 0.143 CUT-OFF / 粒子像の数: 78547 / 対称性のタイプ: POINT |