ジャーナル: EMBO Rep / 年: 2017 タイトル: Architecture of the yeast Elongator complex. 著者: Maria I Dauden / Jan Kosinski / Olga Kolaj-Robin / Ambroise Desfosses / Alessandro Ori / Celine Faux / Niklas A Hoffmann / Osita F Onuma / Karin D Breunig / Martin Beck / Carsten Sachse / ...著者: Maria I Dauden / Jan Kosinski / Olga Kolaj-Robin / Ambroise Desfosses / Alessandro Ori / Celine Faux / Niklas A Hoffmann / Osita F Onuma / Karin D Breunig / Martin Beck / Carsten Sachse / Bertrand Séraphin / Sebastian Glatt / Christoph W Müller / 要旨: The highly conserved eukaryotic Elongator complex performs specific chemical modifications on wobble base uridines of tRNAs, which are essential for proteome stability and homeostasis. The complex is ...The highly conserved eukaryotic Elongator complex performs specific chemical modifications on wobble base uridines of tRNAs, which are essential for proteome stability and homeostasis. The complex is formed by six individual subunits (Elp1-6) that are all equally important for its tRNA modification activity. However, its overall architecture and the detailed reaction mechanism remain elusive. Here, we report the structures of the fully assembled yeast Elongator and the Elp123 sub-complex solved by an integrative structure determination approach showing that two copies of the Elp1, Elp2, and Elp3 subunits form a two-lobed scaffold, which binds Elp456 asymmetrically. Our topological models are consistent with previous studies on individual subunits and further validated by complementary biochemical analyses. Our study provides a structural framework on how the tRNA modification activity is carried out by Elongator.
ダウンロード / ファイル: emd_4153.map.gz / 形式: CCP4 / 大きさ: 42.9 MB / タイプ: IMAGE STORED AS FLOATING POINT NUMBER (4 BYTES)
注釈
Partial Elp123 sub-complex from S.cerevisiae in which one of the Elp2 subunits is missing.
ボクセルのサイズ
X=Y=Z: 2.2 Å
密度
表面レベル
登録者による: 0.0284 / ムービー #1: 0.0284
最小 - 最大
-0.043942522 - 0.10950313
平均 (標準偏差)
0.000026676707 (±0.005108263)
対称性
空間群: 1
詳細
EMDB XML:
マップ形状
Axis order
X
Y
Z
Origin
0
0
0
サイズ
224
224
224
Spacing
224
224
224
セル
A=B=C: 492.80002 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
2.2
2.2
2.2
M x/y/z
224
224
224
origin x/y/z
0.000
0.000
0.000
length x/y/z
492.800
492.800
492.800
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
224
224
224
D min/max/mean
-0.044
0.110
0.000
-
添付データ
-
試料の構成要素
-
全体 : S.cerevisiae partial Elp123 sub-complex, without one of the Elp2 ...
全体
名称: S.cerevisiae partial Elp123 sub-complex, without one of the Elp2 subunits.
要素
複合体: S.cerevisiae partial Elp123 sub-complex, without one of the Elp2 subunits.
-
超分子 #1: S.cerevisiae partial Elp123 sub-complex, without one of the Elp2 ...
超分子
名称: S.cerevisiae partial Elp123 sub-complex, without one of the Elp2 subunits. タイプ: complex / ID: 1 / 親要素: 0 詳細: Composed of two copies of Elp1 and Elp3 and only one copy of Elp2.
由来(天然)
生物種: Saccharomyces cerevisiae (パン酵母) / 株: BS1173
分子量
理論値: 530 KDa
-
実験情報
-
構造解析
手法
ネガティブ染色法
解析
単粒子再構成法
試料の集合状態
particle
-
試料調製
濃度
0.035 mg/mL
緩衝液
pH: 7.6 構成要素:
濃度
式
名称
10.0 mM
C8H18N2O4S
HEPES buffer
125.0 mM
NaCl
Sodium chloride
5.0 mM
C2H6OS
2-mercaptoethanol
染色
タイプ: NEGATIVE / 材質: Uranyl Acetate 詳細: 3.5 ul aliquots of freshly purified Elp123 complex were applied to glow discharged carbon copper-collodion (Sigma) grids for 2 min and stained with a 1% uranyl acetate solution (w/v)