caveolin-mediated endocytosis of virus by host cell / T=7 icosahedral viral capsid / host cell nucleus / virion attachment to host cell / structural molecule activity 類似検索 - 分子機能
Capsid protein VP1,Polyomavirus / Polyomavirus capsid protein VP1 superfamily / Polyomavirus coat protein / Double-stranded DNA virus, group I, capsid 類似検索 - ドメイン・相同性
Capsid protein VP1 / Capsid protein VP1 類似検索 - 構成要素
National Institutes of Health/National Institute of Neurological Disorders and Stroke (NIH/NINDS)
R01NS088367
米国
National Institutes of Health/National Institute of Neurological Disorders and Stroke (NIH/NINDS)
R01NS092662
米国
National Institutes of Health/National Institute Of Allergy and Infectious Diseases (NIH/NIAID)
R01AI107121
米国
National Institutes of Health/National Institute of Neurological Disorders and Stroke (NIH/NINDS)
F32NS106730
米国
National Institutes of Health/National Institute of Neurological Disorders and Stroke (NIH/NINDS)
F31NS083336
米国
National Institutes of Health/National Cancer Institute (NIH/NCI)
T32CA060395
米国
引用
ジャーナル: Elife / 年: 2020 タイトル: Antibody escape by polyomavirus capsid mutation facilitates neurovirulence. 著者: Matthew D Lauver / Daniel J Goetschius / Colleen S Netherby-Winslow / Katelyn N Ayers / Ge Jin / Daniel G Haas / Elizabeth L Frost / Sung Hyun Cho / Carol M Bator / Stephanie M Bywaters / ...著者: Matthew D Lauver / Daniel J Goetschius / Colleen S Netherby-Winslow / Katelyn N Ayers / Ge Jin / Daniel G Haas / Elizabeth L Frost / Sung Hyun Cho / Carol M Bator / Stephanie M Bywaters / Neil D Christensen / Susan L Hafenstein / Aron E Lukacher / 要旨: JCPyV polyomavirus, a member of the human virome, causes progressive multifocal leukoencephalopathy (PML), an oft-fatal demyelinating brain disease in individuals receiving immunomodulatory therapies. ...JCPyV polyomavirus, a member of the human virome, causes progressive multifocal leukoencephalopathy (PML), an oft-fatal demyelinating brain disease in individuals receiving immunomodulatory therapies. Mutations in the major viral capsid protein, VP1, are common in JCPyV from PML patients (JCPyV-PML) but whether they confer neurovirulence or escape from virus-neutralizing antibody (nAb) in vivo is unknown. A mouse polyomavirus (MuPyV) with a sequence-equivalent JCPyV-PML VP1 mutation replicated poorly in the kidney, a major reservoir for JCPyV persistence, but retained the CNS infectivity, cell tropism, and neuropathology of the parental virus. This mutation rendered MuPyV resistant to a monoclonal Ab (mAb), whose specificity overlapped the endogenous anti-VP1 response. Using cryo-EM and a custom sub-particle refinement approach, we resolved an MuPyV:Fab complex map to 3.2 Å resolution. The structure revealed the mechanism of mAb evasion. Our findings demonstrate convergence between nAb evasion and CNS neurovirulence in vivo by a frequent JCPyV-PML VP1 mutation.
中空か: NO / エンベロープを持つか: NO / 単離: STRAIN / タイプ: VIRION
ウイルス殻
直径: 450 nm / 三角数 (T数): 7
緩衝液
pH: 7.9 詳細: 10 mM HEPES pH 7.9, 1 mM CaCl2, 1 mM MgCl2, 5 mM KCl
緩衝液成分
ID
濃度
式
Buffer-ID
1
10mM
HEPES
1
2
1mM
CaCl2
1
3
1mM
MgCl2
1
4
5mM
KCl
1
試料
濃度: 2.8 mg/ml / 包埋: NO / シャドウイング: NO / 染色: NO / 凍結: YES / 詳細: MuPyV (2.8 mg/mL)
急速凍結
凍結剤: ETHANE / 湿度: 95 %
-
電子顕微鏡撮影
実験機器
モデル: Titan Krios / 画像提供: FEI Company
顕微鏡
モデル: FEI TITAN KRIOS
電子銃
電子線源: FIELD EMISSION GUN / 加速電圧: 300 kV / 照射モード: OTHER
電子レンズ
モード: BRIGHT FIELD
撮影
電子線照射量: 45 e/Å2 フィルム・検出器のモデル: FEI FALCON III (4k x 4k)
-
解析
ソフトウェア
名称
バージョン
分類
phenix.real_space_refine
1.17.1-3660_3260
精密化
PHENIX
1.17.1-3660_3260
精密化
EMソフトウェア
ID
名称
バージョン
カテゴリ
4
cryoSPARC
CTF補正
7
UCSF Chimera
モデルフィッティング
9
Coot
モデル精密化
10
PHENIX
モデル精密化
11
RELION
3.1
初期オイラー角割当
12
RELION
3.1
最終オイラー角割当
14
RELION
3.1
3次元再構成
CTF補正
タイプ: PHASE FLIPPING AND AMPLITUDE CORRECTION
3次元再構成
解像度: 2.9 Å / 解像度の算出法: FSC 0.143 CUT-OFF / 粒子像の数: 929940 / 対称性のタイプ: POINT
原子モデル構築
空間: REAL 詳細: Homology model for Fab was generated using SwissModel. Initial models were docked into density in Chimera. Fab CDR loops were manually rebuilt in Coot. Iterative rounds of real space ...詳細: Homology model for Fab was generated using SwissModel. Initial models were docked into density in Chimera. Fab CDR loops were manually rebuilt in Coot. Iterative rounds of real space refinements (PHENIX) and manual adjustment (coot) were conducted to improve fit to density.