Functional characterization of XendoU, the endoribonuclease involved in small nucleolar RNA biosynthesis

被引:47
作者
Gioia, U
Laneve, P
Dlakic, M
Arceci, M
Bozzoni, I
Caffarelli, E
机构
[1] Univ Roma La Sapienza, Dept Genet & Mol Biol, Fdn Cenci Bolognetti, Inst Pasteur, I-00185 Rome, Italy
[2] CNR, Inst Mol Biol & Pathol, I-00185 Rome, Italy
[3] Montana State Univ, Dept Microbiol, Bozeman, MT 59717 USA
关键词
D O I
10.1074/jbc.M501160200
中图分类号
Q5 [生物化学]; Q7 [分子生物学];
学科分类号
071010 ; 081704 ;
摘要
XendoU is the endoribonuclease involved in the biosynthesis of a specific subclass of Xenopus laevis intron-encoded small nucleolar RNAs. XendoU has no homology to any known cellular RNase, although it has sequence similarity with proteins tentatively annotated as serine proteases. It has been recently shown that XendoU represents the cellular counterpart of a nidovirus replicative endoribonuclease (NendoU), which plays a critical role in viral replication and transcription. In this paper, we combined prediction and experimental data to define the amino acid residues directly involved in XendoU catalysis. Specifically, we find that XendoU residues Glu-161, Glu-167, His-162, His-178, and Lys-224 are essential for RNA cleavage, which occurs in the presence of manganese ions. Furthermore, we identified the RNA sequence required for XendoU binding and showed that the formation of XendoU-RNA complex is Mn2+-independent.
引用
收藏
页码:18996 / 19002
页数:7
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