+データを開く
-基本情報
登録情報 | データベース: EMDB / ID: EMD-2424 | |||||||||
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タイトル | Sub-tomogram average of the glycoprotein spike of Boid Inclusion Body Disease associated ArenaVirus | |||||||||
マップデータ | Sub-tomogram reconstruction of the glycoprotein spike from Boid Inclusion Body Disease-associated arenavirus | |||||||||
試料 |
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キーワード | Glycoprotein Trimer | |||||||||
生物種 | Arenavirus (ウイルス) | |||||||||
手法 | サブトモグラム平均法 / クライオ電子顕微鏡法 / 解像度: 32.0 Å | |||||||||
データ登録者 | Hetzel U / Sironen T / Laurinmaki P / Liljeroos L / Patjas A / Henttonen H / Vaheri A / Artelt A / Kipar A / Butcher SJ ...Hetzel U / Sironen T / Laurinmaki P / Liljeroos L / Patjas A / Henttonen H / Vaheri A / Artelt A / Kipar A / Butcher SJ / Vapalahti O / Hepojoki J | |||||||||
引用 | ジャーナル: J Virol / 年: 2013 タイトル: Isolation, identification, and characterization of novel arenaviruses, the etiological agents of boid inclusion body disease. 著者: Udo Hetzel / Tarja Sironen / Pasi Laurinmäki / Lassi Liljeroos / Aino Patjas / Heikki Henttonen / Antti Vaheri / Annette Artelt / Anja Kipar / Sarah J Butcher / Olli Vapalahti / Jussi Hepojoki / 要旨: Boid inclusion body disease (BIBD) is a progressive, usually fatal disease of constrictor snakes, characterized by cytoplasmic inclusion bodies (IB) in a wide range of cell types. To identify the ...Boid inclusion body disease (BIBD) is a progressive, usually fatal disease of constrictor snakes, characterized by cytoplasmic inclusion bodies (IB) in a wide range of cell types. To identify the causative agent of the disease, we established cell cultures from BIBD-positive and -negative boa constrictors. The IB phenotype was maintained in cultured cells of affected animals, and supernatants from these cultures caused the phenotype in cultures originating from BIBD-negative snakes. Viruses were purified from the supernatants by ultracentrifugation and subsequently identified as arenaviruses. Purified virus also induced the IB phenotype in naive cells, which fulfilled Koch's postulates in vitro. One isolate, tentatively designated University of Helsinki virus (UHV), was studied in depth. Sequencing confirmed that UHV is a novel arenavirus species that is distinct from other known arenaviruses including those recently identified in snakes with BIBD. The morphology of UHV was established by cryoelectron tomography and subtomographic averaging, revealing the trimeric arenavirus spike structure at 3.2-nm resolution. Immunofluorescence, immunohistochemistry, and immunoblotting with a polyclonal rabbit antiserum against UHV and reverse transcription-PCR (RT-PCR) revealed the presence of genetically diverse arenaviruses in a large cohort of snakes with BIBD, confirming the causative role of arenaviruses. Some snakes were also found to carry arenavirus antibodies. Furthermore, mammalian cells (Vero E6) were productively infected with UHV, demonstrating the potential of arenaviruses to cross species barriers. In conclusion, we propose the newly identified lineage of arenaviruses associated with BIBD as a novel taxonomic entity, boid inclusion body disease-associated arenaviruses (BIBDAV), in the family Arenaviridae. | |||||||||
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 | EMマップ: SurfViewMolmilJmol/JSmol |
添付画像 |
-ダウンロードとリンク
-EMDBアーカイブ
マップデータ | emd_2424.map.gz | 900.1 KB | EMDBマップデータ形式 | |
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ヘッダ (付随情報) | emd-2424-v30.xml emd-2424.xml | 9.4 KB 9.4 KB | 表示 表示 | EMDBヘッダ |
画像 | emd_2424.tif | 158.1 KB | ||
アーカイブディレクトリ | http://ftp.pdbj.org/pub/emdb/structures/EMD-2424 ftp://ftp.pdbj.org/pub/emdb/structures/EMD-2424 | HTTPS FTP |
-検証レポート
文書・要旨 | emd_2424_validation.pdf.gz | 215.5 KB | 表示 | EMDB検証レポート |
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文書・詳細版 | emd_2424_full_validation.pdf.gz | 214.6 KB | 表示 | |
XML形式データ | emd_2424_validation.xml.gz | 5.5 KB | 表示 | |
アーカイブディレクトリ | https://ftp.pdbj.org/pub/emdb/validation_reports/EMD-2424 ftp://ftp.pdbj.org/pub/emdb/validation_reports/EMD-2424 | HTTPS FTP |
-関連構造データ
-リンク
EMDBのページ | EMDB (EBI/PDBe) / EMDataResource |
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-マップ
ファイル | ダウンロード / ファイル: emd_2424.map.gz / 形式: CCP4 / 大きさ: 1001 KB / タイプ: IMAGE STORED AS FLOATING POINT NUMBER (4 BYTES) | ||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
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注釈 | Sub-tomogram reconstruction of the glycoprotein spike from Boid Inclusion Body Disease-associated arenavirus | ||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
投影像・断面図 | 画像のコントロール
画像は Spider により作成 | ||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
ボクセルのサイズ | X=Y=Z: 7.6 Å | ||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
密度 |
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対称性 | 空間群: 1 | ||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
詳細 | EMDB XML:
CCP4マップ ヘッダ情報:
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-添付データ
-試料の構成要素
-全体 : Boid inclusion body disease associated arenavirus
全体 | 名称: Boid inclusion body disease associated arenavirus |
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要素 |
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-超分子 #1000: Boid inclusion body disease associated arenavirus
超分子 | 名称: Boid inclusion body disease associated arenavirus / タイプ: sample / ID: 1000 / Number unique components: 1 |
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-分子 #1: glycoprotein C
分子 | 名称: glycoprotein C / タイプ: protein_or_peptide / ID: 1 / Name.synonym: GPC 詳細: The GPC monomer has a predicted weight of 45 after cleavage of the signal sequence. In most arenaviruses, GPC is cleaved into GP1 (24.1 kDa) and GP2 (20.9 kDa). 集合状態: Trimer / 組換発現: No |
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由来(天然) | 生物種: Arenavirus (ウイルス) / 株: University of Helsinki virus 1 |
分子量 | 理論値: 45 KDa |
-実験情報
-構造解析
手法 | クライオ電子顕微鏡法 |
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解析 | サブトモグラム平均法 |
試料の集合状態 | particle |
-試料調製
緩衝液 | pH: 7.4 / 詳細: 0.2M phosphate-buffered saline |
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グリッド | 詳細: C-flat 2/2-4C holey carbon copper grid, glow discharged in air |
凍結 | 凍結剤: ETHANE / 装置: HOMEMADE PLUNGER / 手法: Blot for 3 seconds before plunging |
-電子顕微鏡法
顕微鏡 | FEI TECNAI F20 |
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日付 | 2012年10月28日 |
撮影 | カテゴリ: CCD フィルム・検出器のモデル: GATAN ULTRASCAN 4000 (4k x 4k) 詳細: Data collected with serialEM |
電子線 | 加速電圧: 200 kV / 電子線源: FIELD EMISSION GUN |
電子光学系 | 倍率(補正後): 39400 / 照射モード: FLOOD BEAM / 撮影モード: BRIGHT FIELD / Cs: 2.0 mm / 最大 デフォーカス(公称値): 6.0 µm / 最小 デフォーカス(公称値): 3.0 µm / 倍率(公称値): 39400 |
試料ステージ | 試料ホルダー: Eucentric / 試料ホルダーモデル: GATAN LIQUID NITROGEN / Tilt series - Axis1 - Min angle: -60 ° / Tilt series - Axis1 - Max angle: 60 ° |
実験機器 | モデル: Tecnai F20 / 画像提供: FEI Company |
-画像解析
詳細 | Subtomograms were manually selected from the tomograms by visual inspection |
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最終 再構成 | 想定した対称性 - 点群: C3 (3回回転対称) / アルゴリズム: OTHER / 解像度のタイプ: BY AUTHOR / 解像度: 32.0 Å / 解像度の算出法: FSC 0.5 CUT-OFF / ソフトウェア - 名称: IMOD, PEET, Bsoft 詳細: Final maps were calculated from viruses present in 11 different tilt series. Data were split into two halves at the start of the project and refined separately with independent models. The ...詳細: Final maps were calculated from viruses present in 11 different tilt series. Data were split into two halves at the start of the project and refined separately with independent models. The FSC was calculated from the comparison of these two independent models. 使用したサブトモグラム数: 2463 |