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.
名称: S.cerevisiae Elp123 sub-complex / タイプ: complex / ID: 1 / 親要素: 0 詳細: Composed of two copies of its three subunits: Elp1, Elp2 and Elp3.
由来(天然)
生物種: Saccharomyces cerevisiae (パン酵母) / 株: BS1173
分子量
理論値: 621 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)