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基本情報
登録情報 | データベース: EMDB / ID: EMD-6560 | |||||||||
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タイトル | Bacteriophage phi29 prohead particle stalled during DNA packaging | |||||||||
![]() | Reconstruction of bacteriophage phi29 stalled during DNA packaging via the addition of gamma-S-ATP | |||||||||
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![]() | bacteriophage phi29 / ATPase / DNA packaging motor | |||||||||
生物種 | ![]() ![]() | |||||||||
手法 | 単粒子再構成法 / クライオ電子顕微鏡法 / 解像度: 12.0 Å | |||||||||
![]() | Mao H / Saha M / Reyes-Aldrete E / Sherman M / Woodson M / Jardine PJ / Grimes S / Morais M | |||||||||
![]() | ![]() タイトル: Structural and Molecular Basis for Coordination in a Viral DNA Packaging Motor. 著者: Huzhang Mao / Mitul Saha / Emilio Reyes-Aldrete / Michael B Sherman / Michael Woodson / Rockney Atz / Shelley Grimes / Paul J Jardine / Marc C Morais / ![]() 要旨: Ring NTPases are a class of ubiquitous molecular motors involved in basic biological partitioning processes. dsDNA viruses encode ring ATPases that translocate their genomes to near-crystalline ...Ring NTPases are a class of ubiquitous molecular motors involved in basic biological partitioning processes. dsDNA viruses encode ring ATPases that translocate their genomes to near-crystalline densities within pre-assembled viral capsids. Here, X-ray crystallography, cryoEM, and biochemical analyses of the dsDNA packaging motor in bacteriophage phi29 show how individual subunits are arranged in a pentameric ATPase ring and suggest how their activities are coordinated to translocate dsDNA. The resulting pseudo-atomic structure of the motor and accompanying functional analyses show how ATP is bound in the ATPase active site; identify two DNA contacts, including a potential DNA translocating loop; demonstrate that a trans-acting arginine finger is involved in coordinating hydrolysis around the ring; and suggest a functional coupling between the arginine finger and the DNA translocating loop. The ability to visualize the motor in action illuminates how the different motor components interact with each other and with their DNA substrate. | |||||||||
履歴 |
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構造の表示
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構造ビューア | EMマップ: ![]() ![]() ![]() |
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マップデータ | ![]() | 29.8 MB | ![]() | |
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ヘッダ (付随情報) | ![]() ![]() | 10.4 KB 10.4 KB | 表示 表示 | ![]() |
画像 | ![]() ![]() | 64.5 KB 3.8 KB | ||
アーカイブディレクトリ | ![]() ![]() | HTTPS FTP |
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文書・要旨 | ![]() | 78.1 KB | 表示 | ![]() |
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文書・詳細版 | ![]() | 77.2 KB | 表示 | |
XML形式データ | ![]() | 493 B | 表示 | |
アーカイブディレクトリ | ![]() ![]() | HTTPS FTP |
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注釈 | Reconstruction of bacteriophage phi29 stalled during DNA packaging via the addition of gamma-S-ATP | ||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
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: 1.94 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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試料の構成要素
-全体 : Reconstruction of bacteriophage phi29 stalled during DNA packagin...
全体 | 名称: Reconstruction of bacteriophage phi29 stalled during DNA packaging via the addition of gamma-S-ATP. Only particles that had clearly packaged DNA were included in the reconstruction. |
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要素 |
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-超分子 #1000: Reconstruction of bacteriophage phi29 stalled during DNA packagin...
超分子 | 名称: Reconstruction of bacteriophage phi29 stalled during DNA packaging via the addition of gamma-S-ATP. Only particles that had clearly packaged DNA were included in the reconstruction. タイプ: sample / ID: 1000 集合状態: The particle is based on a prolate icosahedron with T = 3, Q = 5 quasi-symmetry wherein one pentameric capsomer at one end of the particle is replaced by a dodecameric gp10 connector ...集合状態: The particle is based on a prolate icosahedron with T = 3, Q = 5 quasi-symmetry wherein one pentameric capsomer at one end of the particle is replaced by a dodecameric gp10 connector protein and a pentameric pRNA and gp16 ATPase ring. Number unique components: 1 |
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-超分子 #1: Bacillus phage phi29
超分子 | 名称: Bacillus phage phi29 / タイプ: virus / ID: 1 / Name.synonym: bacteriophage phi29 詳細: Reconstruction of bacteriophage phi29 stalled during DNA packaging via the addition of gamma-S-ATP. After two minutes incubation, DNAses were added to remove unpackaged DNA. Only particles ...詳細: Reconstruction of bacteriophage phi29 stalled during DNA packaging via the addition of gamma-S-ATP. After two minutes incubation, DNAses were added to remove unpackaged DNA. Only particles that had clearly packaged were included in the reconstruction. NCBI-ID: 10756 / 生物種: Bacillus phage phi29 / データベース: NCBI / ウイルスタイプ: VIRUS-LIKE PARTICLE / ウイルス・単離状態: SPECIES / ウイルス・エンベロープ: No / ウイルス・中空状態: Yes / Syn species name: bacteriophage phi29 |
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宿主 | 生物種: ![]() ![]() |
-実験情報
-構造解析
手法 | クライオ電子顕微鏡法 |
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![]() | 単粒子再構成法 |
試料の集合状態 | particle |
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試料調製
緩衝液 | pH: 7.8 / 詳細: 25 mM Tris-HCl, 5 mM MgCl2, 50 mM NaCl |
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グリッド | 詳細: Quantifoil holey carbon on top of 200 mesh copper grid, plasma-cleaned |
凍結 | 凍結剤: ETHANE / 装置: HOMEMADE PLUNGER 手法: Blot from behind the sample for 4 seconds before plunging. |
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電子顕微鏡法
顕微鏡 | JEOL 2200FS |
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アライメント法 | Legacy - 非点収差: Objective lens astigmatism was corrected at 100,000 times magnification. |
特殊光学系 | エネルギーフィルター - 名称: JEOL |
日付 | 2013年6月24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) 平均電子線量: 25 e/Å2 |
電子線 | 加速電圧: 200 kV / 電子線源: ![]() |
電子光学系 | 倍率(補正後): 60000 / 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): 3.0 µm / 最小 デフォーカス(公称値): 0.5 µm / 倍率(公称値): 60000 |
試料ステージ | 試料ホルダー: Side entry liquid nitrogen-cooled cryo specimen holder 試料ホルダーモデル: GATAN LIQUID NITROGEN |
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画像解析
詳細 | The particles were selected using semi-automatic particle selection, followed by manual deletion of bad particles and manual selection of missed particles. |
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CTF補正 | 詳細: Each Micrograph |
最終 再構成 | アルゴリズム: OTHER / 解像度のタイプ: BY AUTHOR / 解像度: 12.0 Å / 解像度の算出法: OTHER / ソフトウェア - 名称: EMAN 詳細: C5 symmetry was imposed in all stages of the reconstruction, with the 5-fold axis along Z. The final map was low-pass filtered at 14 Angstrom. 使用した粒子像数: 1871 |