Solution structure and dynamics of yeast elongin C in complex with a von Hippel-Lindau peptide
Solution structure and dynamics of yeast elongin C in complex with a von Hippel-Lindau peptide
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DOI：
10.1006/jmbi.2001.4938
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发表时间：
2001-09-07
影响因子：
5.6
通讯作者：
Arrowsmith, CH
中科院分区：
文献类型：
--
作者：
Botuyan, MV;Mer, G;Arrowsmith, CH
Elongin是一种转录延伸因子，通过抑制RNA聚合酶II在DNA沿线许多位置的短暂停顿来刺激延伸速度。它在哺乳动物中是异源三聚体，由细长蛋白A、B和C亚基组成，与一类被称为SCF(Skp1-CDc53(Cullin)-F-box)的E3泛素连接酶具有总体相似性。细长蛋白B和C的亚复合体是von Hippel-Lindau(VHL)肿瘤抑制蛋白负调控的靶点。酿酒酵母Elongin C与哺乳动物细长蛋白C具有很高的序列相似性，我们用核磁共振波谱测定了Elc1与人VHL多肽VHL(157-171)形成的复合体的三维结构，代表Elc1的主要结合部位。结合的VHL多肽是完全螺旋的。Elc1利用两个C-末端螺旋和一个中间环形成适合VHL(157-171)的结合槽。化学位移微扰和动力学分析表明，Elc1/VHL(157-171)络合物的形成伴随着全球构象的变化。此外，VHL多肽结合后Elc1在微秒到毫秒时间尺度上的构象交换现象的消失表明了缓慢的内部运动在配体识别中的作用。(C)2001年学术出版社。
Elongin is a transcription elongation factor that stimulates the rate of elongation by suppressing transient pausing by RNA polymerase II at many sites along the DNA. It is heterotrimeric in mammals, consisting of elongins A, B and C subunits, and bears overall similarity to a class of E3 ubiquitin ligases known as SCF (Skp1-Cdc53 (cullin)-F-box) complexes. A subcomplex of elongins B and C is a target for negative regulation by the von Hippel-Lindau (VHL) tumor-suppressor protein. Elongin C from Saccharomyces cerevisiae, Elc1, exhibits high sequence similarity to mammalian elongin C. Using NMR spectroscopy we have determined the three-dimensional structure of Elc1 in complex with a human VHL peptide, VHL(157-171), representing the major Elc1 binding site. The bound VHL peptide is entirely helical. Elc1 utilizes two C-terminal helices and an intervening loop to form a binding groove that fits VHL(157-171). Chemical shift perturbation and dynamics analyses reveal that a global conformational change accompanies Elc1/VHL(157-171) complex formation. Moreover, the disappearance of conformational exchange phenomena on the microsecond to millisecond time scale within Elc1 upon VHL peptide binding suggests a role for slow internal motions in ligand recognition. (C) 2001 Academic Press.