ジャーナル: Structure / 年: 2013 タイトル: Cryo-electron microscopy three-dimensional structure of the jumbo phage ΦRSL1 infecting the phytopathogen Ralstonia solanacearum. 著者: Grégory Effantin / Ryosuke Hamasaki / Takeru Kawasaki / Maria Bacia / Christine Moriscot / Winfried Weissenhorn / Takashi Yamada / Guy Schoehn / 要旨: ϕRSL1 jumbo phage belongs to a new class of viruses within the Myoviridae family. Here, we report its three-dimensional structure determined by electron cryo microscopy. The icosahedral capsid, the ...ϕRSL1 jumbo phage belongs to a new class of viruses within the Myoviridae family. Here, we report its three-dimensional structure determined by electron cryo microscopy. The icosahedral capsid, the tail helical portion, and the complete tail appendage were reconstructed separately to resolutions of 9 Å, 9 Å, and 28 Å, respectively. The head is rather complex and formed by at least five different proteins, whereas the major capsid proteins resemble those from HK97, despite low sequence conservation. The helical tail structure demonstrates its close relationship to T4 sheath proteins and provides evidence for an evolutionary link of the inner tail tube to the bacterial type VI secretion apparatus. Long fibers extend from the collar region, and their length is consistent with reaching the host cell surface upon tail contraction. Our structural analyses indicate that ϕRSL1 is an unusual member of the Myoviridae that employs conserved protein machines related to different phages and bacteria.
ダウンロード / ファイル: emd_2244.map.gz / 形式: CCP4 / 大きさ: 6.4 MB / タイプ: IMAGE STORED AS FLOATING POINT NUMBER (4 BYTES)
注釈
Reconstruction of the helical trunk of phirsl1 tail. A Bfactor of -538 has been applied to the reconstruction as well as a soft-edged mask.
ボクセルのサイズ
X=Y=Z: 2.26 Å
密度
表面レベル
登録者による: 0.007 / ムービー #1: 0.007
最小 - 最大
-0.02692472 - 0.0357494
平均 (標準偏差)
0.00069069 (±0.00452792)
対称性
空間群: 1
詳細
EMDB XML:
マップ形状
Axis order
X
Y
Z
Origin
0
-3
-5
サイズ
120
120
120
Spacing
120
120
120
セル
A=B=C: 271.2 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
2.26
2.26
2.26
M x/y/z
120
120
120
origin x/y/z
0.000
0.000
0.000
length x/y/z
271.200
271.200
271.200
α/β/γ
90.000
90.000
90.000
start NX/NY/NZ
0
0
0
NX/NY/NZ
45
45
86
MAP C/R/S
1
2
3
start NC/NR/NS
-3
0
-5
NC/NR/NS
120
120
120
D min/max/mean
-0.027
0.036
0.001
-
添付データ
-
試料の構成要素
-
全体 : phirsl1 helical portion of the tail
全体
名称: phirsl1 helical portion of the tail
要素
試料: phirsl1 helical portion of the tail
タンパク質・ペプチド: Tail tube protein
タンパク質・ペプチド: Tail sheath protein
-
超分子 #1000: phirsl1 helical portion of the tail
超分子
名称: phirsl1 helical portion of the tail / タイプ: sample / ID: 1000 / Number unique components: 2
-
分子 #1: Tail tube protein
分子
名称: Tail tube protein / タイプ: protein_or_peptide / ID: 1 / 詳細: UNP B2ZY00 / 組換発現: No / データベース: NCBI
由来(天然)
生物種: Ralstonia phage RSL1 (ファージ)
分子量
理論値: 18 KDa
-
分子 #2: Tail sheath protein
分子
名称: Tail sheath protein / タイプ: protein_or_peptide / ID: 2 / 詳細: UNP B2ZXZ8 / 組換発現: No / データベース: NCBI
由来(天然)
生物種: Ralstonia phage RSL1 (ファージ)
分子量
理論値: 68 KDa
配列
InterPro: Tail sheath protein
-
実験情報
-
構造解析
手法
ネガティブ染色法, クライオ電子顕微鏡法
解析
らせん対称体再構成法
試料の集合状態
filament
-
試料調製
緩衝液
pH: 7.5 / 詳細: 50 mM Tris-HCl [pH 7.5], 100 mM NaCl, 10 mM MgSO4
染色
タイプ: NEGATIVE 詳細: Four microliters of the bacteriophage sample (~0.1 mg/ml) were loaded between the mica-carbon interface. The sample was stained using 2% ammonium molybdate pH 7.5 and air-dried.
凍結
凍結剤: ETHANE / チャンバー内湿度: 100 % / 装置: FEI VITROBOT MARK IV
-
電子顕微鏡法
顕微鏡
FEI POLARA 300
アライメント法
Legacy - 非点収差: Objective lens astigmatism was corrected at 59000