- EMDB-0566: Subtomogram average of the Legionella pneumophila Dot/Icm type IV... -
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基本情報
登録情報
データベース: EMDB / ID: EMD-0566
タイトル
Subtomogram average of the Legionella pneumophila Dot/Icm type IV secretion system with DotF fused to a superfolder GFP (aligning the OM-complex)
マップデータ
Subtomogram average of the Legionella pneumophila Dot/Icm type IV secretion system with DotF fused to a superfolder GFP (aligning the OM-complex).
試料
複合体: Subtomogram average of the Legionella pneumophila Dot/Icm type IV secretion system with DotF fused to a superfolder GFP (aligning the OM-complex).
National Institutes of Health/National Institute Of Allergy and Infectious Diseases
R01AI127401
米国
National Institutes of Health/National Institute Of Allergy and Infectious Diseases
R01AI48052
米国
引用
ジャーナル: Nat Microbiol / 年: 2019 タイトル: Molecular architecture, polar targeting and biogenesis of the Legionella Dot/Icm T4SS. 著者: Debnath Ghosal / Kwangcheol C Jeong / Yi-Wei Chang / Jacob Gyore / Lin Teng / Adam Gardner / Joseph P Vogel / Grant J Jensen / 要旨: Legionella pneumophila survives and replicates inside host cells by secreting ~300 effectors through the defective in organelle trafficking (Dot)/intracellular multiplication (Icm) type IVB secretion ...Legionella pneumophila survives and replicates inside host cells by secreting ~300 effectors through the defective in organelle trafficking (Dot)/intracellular multiplication (Icm) type IVB secretion system (T4BSS). Here, we used complementary electron cryotomography and immunofluorescence microscopy to investigate the molecular architecture and biogenesis of the Dot/Icm secretion apparatus. Electron cryotomography mapped the location of the core and accessory components of the Legionella core transmembrane subcomplex, revealing a well-ordered central channel that opens into a large, windowed secretion chamber with an unusual 13-fold symmetry. Immunofluorescence microscopy deciphered an early-stage assembly process that begins with the targeting of Dot/Icm components to the bacterial poles. Polar targeting of this T4BSS is mediated by two Dot/Icm proteins, DotU and IcmF, that, interestingly, are homologues of the T6SS membrane complex components TssL and TssM, suggesting that the Dot/Icm T4BSS is a hybrid system. Together, these results revealed that the Dot/Icm complex assembles in an 'axial-to-peripheral' pattern.
A: 780.0 Å / B: 936.0 Å / C: 780.0 Å α=β=γ: 90.0 °
CCP4マップ ヘッダ情報:
mode
Image stored as Reals
Å/pix. X/Y/Z
7.8
7.8
7.8
M x/y/z
100
120
100
origin x/y/z
0.000
0.000
0.000
length x/y/z
780.000
936.000
780.000
α/β/γ
90.000
90.000
90.000
MAP C/R/S
1
2
3
start NC/NR/NS
0
0
0
NC/NR/NS
100
120
100
D min/max/mean
-0.000
1.000
0.385
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添付データ
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試料の構成要素
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全体 : Subtomogram average of the Legionella pneumophila Dot/Icm type IV...
全体
名称: Subtomogram average of the Legionella pneumophila Dot/Icm type IV secretion system with DotF fused to a superfolder GFP (aligning the OM-complex).
要素
複合体: Subtomogram average of the Legionella pneumophila Dot/Icm type IV secretion system with DotF fused to a superfolder GFP (aligning the OM-complex).
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超分子 #1: Subtomogram average of the Legionella pneumophila Dot/Icm type IV...
超分子
名称: Subtomogram average of the Legionella pneumophila Dot/Icm type IV secretion system with DotF fused to a superfolder GFP (aligning the OM-complex). タイプ: complex / ID: 1 / 親要素: 0 詳細: We used electron cryotomography and subtomogram averaging to visualize L. pneumophila Dot/Icm (DotF-GFP fusion) complex directly in intact, frozen-hydrated bacterial cells
由来(天然)
生物種: Legionella pneumophila (バクテリア) / 株: Lp02
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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
cell
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試料調製
緩衝液
pH: 6.9
グリッド
詳細: unspecified
凍結
凍結剤: ETHANE-PROPANE
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電子顕微鏡法
顕微鏡
FEI TITAN KRIOS
撮影
フィルム・検出器のモデル: GATAN K2 SUMMIT (4k x 4k) 平均電子線量: 1.2 e/Å2
電子線
加速電圧: 300 kV / 電子線源: FIELD EMISSION GUN
電子光学系
照射モード: FLOOD BEAM / 撮影モード: BRIGHT FIELD
実験機器
モデル: Titan Krios / 画像提供: FEI Company
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画像解析
最終 再構成
想定した対称性 - 点群: C2 (2回回転対称) / 解像度のタイプ: BY AUTHOR / 解像度: 40.0 Å / 解像度の算出法: FSC 0.143 CUT-OFF / ソフトウェア - 名称: IMOD / 使用したサブトモグラム数: 254