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.
全体 : Boid inclusion body disease associated arenavirus
全体
名称: Boid inclusion body disease associated arenavirus
要素
試料: Boid inclusion body disease associated arenavirus
タンパク質・ペプチド: glycoprotein C
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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
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
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実験情報
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構造解析
手法
クライオ電子顕微鏡法
解析
サブトモグラム平均法
試料の集合状態
particle
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試料調製
緩衝液
pH: 7.4 / 詳細: 0.2M phosphate-buffered saline
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
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電子顕微鏡法
顕微鏡
FEI TECNAI F20
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
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
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画像解析
詳細
Subtomograms were manually selected from the tomograms by visual inspection
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