ジャーナル: Nat Microbiol / 年: 2016 タイトル: Platelet-derived growth factor-α receptor is the cellular receptor for human cytomegalovirus gHgLgO trimer. 著者: Anna Kabanova / Jessica Marcandalli / Tongqing Zhou / Siro Bianchi / Ulrich Baxa / Yaroslav Tsybovsky / Daniele Lilleri / Chiara Silacci-Fregni / Mathilde Foglierini / Blanca Maria Fernandez- ...著者: Anna Kabanova / Jessica Marcandalli / Tongqing Zhou / Siro Bianchi / Ulrich Baxa / Yaroslav Tsybovsky / Daniele Lilleri / Chiara Silacci-Fregni / Mathilde Foglierini / Blanca Maria Fernandez-Rodriguez / Aliaksandr Druz / Baoshan Zhang / Roger Geiger / Massimiliano Pagani / Federica Sallusto / Peter D Kwong / Davide Corti / Antonio Lanzavecchia / Laurent Perez / 要旨: Human cytomegalovirus encodes at least 25 membrane glycoproteins that are found in the viral envelope(1). While gB represents the fusion protein, two glycoprotein complexes control the tropism of the ...Human cytomegalovirus encodes at least 25 membrane glycoproteins that are found in the viral envelope(1). While gB represents the fusion protein, two glycoprotein complexes control the tropism of the virus: the gHgLgO trimer is involved in the infection of fibroblasts, and the gHgLpUL128L pentamer is required for infection of endothelial, epithelial and myeloid cells(2-5). Two reports suggested that gB binds to ErbB1 and PDGFRα (refs 6,7); however, these results do not explain the tropism of the virus and were recently challenged(8,9). Here, we provide a 19 Å reconstruction for the gHgLgO trimer and show that it binds with high affinity through the gO subunit to PDGFRα, which is expressed on fibroblasts but not on epithelial cells. We also provide evidence that the trimer is essential for viral entry in both fibroblasts and epithelial cells. Furthermore, we identify the pentamer, which is essential for infection of epithelial cells, as a trigger for the ErbB pathway. These findings help explain the broad tropism of human cytomegalovirus and indicate that PDGFRα and the viral gO subunit could be targeted by novel anti-viral therapies.
生物種: Chinese Hamster Ovary cells / 組換細胞: CHO K1SV / 組換プラスミド: PEE.12.4 and PEE6.4
配列
UniProtKB: Envelope glycoprotein L / GO: viral process, viral envelope / InterPro: Cytomegalovirus glycoprotein L
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実験情報
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構造解析
手法
ネガティブ染色法
解析
単粒子再構成法
試料の集合状態
particle
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試料調製
濃度
0.01 mg/mL
緩衝液
pH: 7.5 / 詳細: 20 mM HEPES, 150 mM NaCl
染色
タイプ: NEGATIVE 詳細: The complex was adsorbed to freshly glow-discharged carbon-coated grids for 15 s, washed with a buffer containing 20 mM HEPES, pH 7.5, and 150 mM NaCl, and stained in several drops of 0.7% uranyl formate
グリッド
詳細: 200 mesh copper grid with carbon support, glow discharged
凍結
凍結剤: NONE / 装置: OTHER
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電子顕微鏡法
顕微鏡
FEI TECNAI 20
アライメント法
Legacy - 非点収差: Objective lens astigmatism was corrected at 100,000 times magnification
試料ホルダー: Room temperature / 試料ホルダーモデル: SIDE ENTRY, EUCENTRIC
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画像解析
詳細
The dataset was subjected to iterative stable alignment and clustering (ISAC) in SPARX. The ISAC classes were then used to generate an initial 3D map in EMAN2, which was used as the initial model for three-dimensional reconstruction and refinement using reference projections in SPIDER.
CTF補正
詳細: Each defocus group
最終 再構成
想定した対称性 - 点群: C1 (非対称) / アルゴリズム: OTHER / 解像度のタイプ: BY AUTHOR / 解像度: 19.3 Å / 解像度の算出法: OTHER / ソフトウェア - 名称: SPARX, EMAN2, SPIDER / 使用した粒子像数: 10356