Accurate placement of substrate RNA by Gar1 in H/ACA RNA-guided   pseudouridylation

doi:10.1093/nar/gkv757

CORC > 北京大学 > 化学与分子工程学院

	Accurate placement of substrate RNA by Gar1 in H/ACA RNA-guided pseudouridylation
	Wang, Peng ; Yang, Lijiang ; Gao, Yi Qin ; Zhao, Xin Sheng
刊名	NUCLEIC ACIDS RESEARCH
	2015
关键词	RIBOSOMAL-RNA CRYSTAL-STRUCTURE SYNTHASE MECHANISM COMPLEX STABILIZATION PROTEINS YEAST RECONSTITUTION TRANSLATION
DOI	10.1093/nar/gkv757
英文摘要	H/ACA RNA-guided ribonucleoprotein particle (RNP), the most complicated RNA pseudouridylase so far known, uses H/ACA guide RNA for substrate capture and four proteins (Cbf5, Nop10, L7Ae and Gar1) for pseudouridylation. Although it was shown that Gar1 not only facilitates the product release, but also enhances the catalytic activity, the chemical role that Gar1 plays in this complicated machinery is largely unknown. Kinetics measurement on Pyrococcus furiosus RNPs at different temperatures making use of fluorescence anisotropy showed that Gar1 reduces the catalytic barrier through affecting the activation entropy instead of enthalpy. Site-directed mutagenesis combined with molecular dynamics simulations demonstrated that V149 in the thumb loop of Cbf5 is critical in placing the target uridine to the right position toward catalytic D85 of Cbf5. The enzyme elegantly aligns the position of uridine in the catalytic site with the help of Gar1. In addition, conversion of uridine to pseudouridine results in a rigid syn configuration of the target nucleotide in the active site and causes Gar1 to pull out the thumb. Both factors guarantee the efficient release of the product.; National Key Basic Research Foundation of China [2010CB912302, 2012CB917304]; National Natural Science Foundation of China [21233002, 91027044, 21373016]; SCI(E); ARTICLE; gaoyq@pku.edu.cn; zhaoxs@pku.edu.cn; 15; 7207-7216; 43
语种	英语
内容类型	期刊论文
源URL	[http://ir.pku.edu.cn/handle/20.500.11897/416379]
专题	化学与分子工程学院
推荐引用方式 GB/T 7714	Wang, Peng,Yang, Lijiang,Gao, Yi Qin,et al. Accurate placement of substrate RNA by Gar1 in H/ACA RNA-guided pseudouridylation[J]. NUCLEIC ACIDS RESEARCH,2015.
APA	Wang, Peng,Yang, Lijiang,Gao, Yi Qin,&Zhao, Xin Sheng.(2015).Accurate placement of substrate RNA by Gar1 in H/ACA RNA-guided pseudouridylation.NUCLEIC ACIDS RESEARCH.
MLA	Wang, Peng,et al."Accurate placement of substrate RNA by Gar1 in H/ACA RNA-guided pseudouridylation".NUCLEIC ACIDS RESEARCH (2015).