期刊
NATURE CHEMICAL BIOLOGY
卷 12, 期 9, 页码 702-+出版社
NATURE PUBLISHING GROUP
DOI: 10.1038/nchembio.2125
关键词
-
资金
- National Institute of General Medical Sciences from US National Institutes of Health (NIH) [P41 GM103403]
- US DOE [DE-AC02-06CH11357]
- Zhejiang University
- Thousand Young Talents Plan of China
- US NIH [1 U19 CA179564]
- Memorial Sloan-Kettering Cancer Center Core Grant [P30 CA008748]
- Austrian Science Fund FWF [P27947, I1040, P26550, P28725]
- Swiss National Foundation SNF
- Austrian Science Fund (FWF) [I 1040, P 27947] Funding Source: researchfish
- Austrian Science Fund (FWF) [P26550, I1040, P27947] Funding Source: Austrian Science Fund (FWF)
The field of small self-cleaving nucleolytic ribozymes has been invigorated by the recent discovery of the twister, twister-sister, pistol and hatchet ribozymes. We report the crystal structure of a pistol ribozyme termed env25, which adopts a compact tertiary architecture stabilized by an embedded pseudoknot fold. The G-U cleavage site adopts a splayed-apart conformation with in-line alignment of the modeled 2'-O of G for attack on the adjacent to-be-cleaved P-O5' bond. Highly conserved residues G40 (N1 position) and A32 (N3 and 2'-OH positions) are aligned to act as a general base and a general acid, respectively, to accelerate cleavage chemistry, with their roles confirmed by cleavage assays on variants, and an increased pK(a) of 4.7 for A32. Our structure of the pistol ribozyme defined how the overall and local topologies dictate the in-line alignment at the G-U cleavage site, with cleavage assays on variants revealing key residues that participate in acid-base-catalyzed cleavage chemistry.
作者
我是这篇论文的作者
点击您的名字以认领此论文并将其添加到您的个人资料中。
推荐
暂无数据