ジャーナル: Nat Microbiol / 年: 2022 タイトル: Identification and structure of an extracellular contractile injection system from the marine bacterium Algoriphagus machipongonensis. 著者: Jingwei Xu / Charles F Ericson / Yun-Wei Lien / Florentine U N Rutaganira / Fabian Eisenstein / Miki Feldmüller / Nicole King / Martin Pilhofer / 要旨: Contractile injection systems (CISs) are phage tail-like nanomachines, mediating bacterial cell-cell interactions as either type VI secretion systems (T6SSs) or extracellular CISs (eCISs). ...Contractile injection systems (CISs) are phage tail-like nanomachines, mediating bacterial cell-cell interactions as either type VI secretion systems (T6SSs) or extracellular CISs (eCISs). Bioinformatic studies uncovered a phylogenetic group of hundreds of putative CIS gene clusters that are highly diverse and widespread; however, only four systems have been characterized. Here we studied a putative CIS gene cluster in the marine bacterium Algoriphagus machipongonensis. Using an integrative approach, we show that the system is compatible with an eCIS mode of action. Our cryo-electron microscopy structure revealed several features that differ from those seen in other CISs: a 'cap adaptor' located at the distal end, a 'plug' exposed to the tube lumen, and a 'cage' formed by massive extensions of the baseplate. These elements are conserved in other CISs, and our genetic tools identified that they are required for assembly, cargo loading and function. Furthermore, our atomic model highlights specific evolutionary hotspots and will serve as a framework for understanding and re-engineering CISs.
ダウンロード / ファイル: emd_13720.map.gz / 形式: CCP4 / 大きさ: 127.4 MB / タイプ: IMAGE STORED AS SIGNED BYTE
ボクセルのサイズ
X=Y=Z: 11 Å
密度
最小 - 最大
-128.0 - 127.0
平均 (標準偏差)
40.966137 (±18.628868)
対称性
空間群: 1
詳細
EMDB XML:
マップ形状
Axis order
X
Y
Z
Origin
0
1
75
サイズ
928
960
150
Spacing
960
928
150
セル
A: 10560.0 Å / B: 10208.0 Å / C: 1650.0 Å α=β=γ: 90.0 °
CCP4マップ ヘッダ情報:
mode
envelope stored as signed bytes (from -128 lowest to 127 highest)
Å/pix. X/Y/Z
11
11
11
M x/y/z
960
928
150
origin x/y/z
0.000
0.000
0.000
length x/y/z
10560.000
10208.000
1650.000
α/β/γ
90.000
90.000
90.000
start NX/NY/NZ
0
0
0
NX/NY/NZ
450
450
450
MAP C/R/S
1
2
3
start NC/NR/NS
1
0
75
NC/NR/NS
960
928
150
D min/max/mean
-128.000
127.000
40.966
-
添付データ
-
試料の構成要素
-
全体 : The purified extracellular contractile injection system (AlgoCIS)...
全体
名称: The purified extracellular contractile injection system (AlgoCIS) Cgo1 deficient mutant
要素
複合体: The purified extracellular contractile injection system (AlgoCIS) Cgo1 deficient mutant
-
超分子 #1: The purified extracellular contractile injection system (AlgoCIS)...
超分子
名称: The purified extracellular contractile injection system (AlgoCIS) Cgo1 deficient mutant タイプ: complex / ID: 1 / 親要素: 0
由来(天然)
生物種: Algoriphagus machipongonensis (バクテリア)
-
実験情報
-
構造解析
手法
クライオ電子顕微鏡法
解析
電子線トモグラフィー法
試料の集合状態
particle
-
試料調製
緩衝液
pH: 7.5
凍結
凍結剤: ETHANE-PROPANE
切片作成
その他: NO SECTIONING
位置合わせマーカー
Manufacturer: cytodiagnostics / 直径: 10 nm
-
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) 検出モード: COUNTING / 平均電子線量: 2.1 e/Å2 詳細: 2.1 electrons for each tilt, while total ~130 electron for one tilt series (61 tilts).