National Institutes of Health/Office of the Director
1DP2GM123494
米国
National Institutes of Health/National Institute of General Medical Sciences
R01-GM057045
米国
引用
ジャーナル: Elife / 年: 2019 タイトル: The molecular architecture of engulfment during sporulation. 著者: Kanika Khanna / Javier Lopez-Garrido / Ziyi Zhao / Reika Watanabe / Yuan Yuan / Joseph Sugie / Kit Pogliano / Elizabeth Villa / 要旨: The study of bacterial cell biology is limited by difficulties in visualizing cellular structures at high spatial resolution within their native milieu. Here, we visualize sporulation using cryo- ...The study of bacterial cell biology is limited by difficulties in visualizing cellular structures at high spatial resolution within their native milieu. Here, we visualize sporulation using cryo-electron tomography coupled with cryo-focused ion beam milling, allowing the reconstruction of native-state cellular sections at molecular resolution. During sporulation, an asymmetrically-positioned septum generates a larger mother cell and a smaller forespore. Subsequently, the mother cell engulfs the forespore. We show that the septal peptidoglycan is not completely degraded at the onset of engulfment. Instead, the septum is uniformly and only slightly thinned as it curves towards the mother cell. Then, the mother cell membrane migrates around the forespore in tiny finger-like projections, whose formation requires the mother cell SpoIIDMP protein complex. We propose that a limited number of SpoIIDMP complexes tether to and degrade the peptidoglycan ahead of the engulfing membrane, generating an irregular membrane front.
ダウンロード / ファイル: emd_20336.map.gz / 形式: CCP4 / 大きさ: 37.5 MB / タイプ: IMAGE STORED AS SIGNED BYTE
注釈
Cryo-electron tomogram of Bacillus subtilis wild type sporangium with curved septum
ボクセルのサイズ
X=Y=Z: 24.48 Å
密度
最小 - 最大
-128 - 127
平均 (標準偏差)
2.5190954 (±6.83525)
対称性
空間群: 1
詳細
EMDB XML:
マップ形状
Axis order
X
Y
Z
Origin
-244
-176
71
サイズ
561
580
121
Spacing
580
561
121
セル
A: 14198.399 Å / B: 13733.279 Å / C: 2962.0798 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
24.479998275862
24.479998217469
24.48
M x/y/z
580
561
121
origin x/y/z
0.000
0.000
0.000
length x/y/z
14198.399
13733.279
2962.080
α/β/γ
90.000
90.000
90.000
start NX/NY/NZ
0
0
0
NX/NY/NZ
162
182
277
MAP C/R/S
1
2
3
start NC/NR/NS
-176
-244
71
NC/NR/NS
580
561
121
D min/max/mean
-128.000
127.000
2.519
-
添付データ
-
試料の構成要素
-
全体 : Bacillus subtilis wild type sporangium with flat septum
全体
名称: Bacillus subtilis wild type sporangium with flat septum
要素
細胞: Bacillus subtilis wild type sporangium with flat septum
-
超分子 #1: Bacillus subtilis wild type sporangium with flat septum
超分子
名称: Bacillus subtilis wild type sporangium with flat septum タイプ: cell / ID: 1 / 親要素: 0
由来(天然)
生物種: Bacillus subtilis PY79 (枯草菌)
-
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
cell
-
試料調製
緩衝液
pH: 7
グリッド
詳細: unspecified
凍結
凍結剤: ETHANE-PROPANE
切片作成
集束イオンビーム - 装置: OTHER / 集束イオンビーム - イオン: OTHER / 集束イオンビーム - 電圧: 30 kV / 集束イオンビーム - 電流: 0.03 nA / 集束イオンビーム - 時間: 3600 sec. / 集束イオンビーム - 温度: 100 K / 集束イオンビーム - Initial thickness: 1200 nm / 集束イオンビーム - 最終 厚さ: 200 nm 集束イオンビーム - 詳細: The value given for _emd_sectioning_focused_ion_beam.instrument is FEI Scios Dual Beam. This is not in a list of allowed values set(['DB235', 'OTHER']) so OTHER is ...集束イオンビーム - 詳細: The value given for _emd_sectioning_focused_ion_beam.instrument is FEI Scios Dual Beam. This is not in a list of allowed values set(['DB235', 'OTHER']) so OTHER is written into the XML file.
-
電子顕微鏡法
顕微鏡
FEI POLARA 300
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: 58.0 e/Å2