A new class of enzyme acting on damaged ribosomes: ribosomal RNA apurinic site specific lyase found in wheat germ

被引:48
作者
Ogasawara, T [1 ]
Sawasaki, T [1 ]
Morishita, R [1 ]
Ozawa, A [1 ]
Madin, K [1 ]
Endo, Y [1 ]
机构
[1] Ehime Univ, Fac Engn, Dept Appl Chem, Matsuyama, Ehime 7908577, Japan
关键词
apurinic site; cleavage; lyase; rRNA; sarcin-ricin domain;
D O I
10.1093/emboj/18.22.6522
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
A new enzyme, which we named ribosomal RNA apurinic site specific lyase (RALyase), is described. The protein was found in wheat embryos and has a molecular weight of 50 625 Da, The enzyme specifically cleaves the phosphodiester bond at the 3' side of the apurinic site introduced by ribosome-inactivating proteins into the sarcin/ricin domain of 28S rRNA, The 3' and 5' ends of wheat 28S rRNA at the cleavage site are 5'-GUACG-alpha-hyroxy-alpha,beta-unsaturated aldehyde and pGAGGA-3', demonstrating that the enzyme catalyzes a p-elimination reaction. The substrate specificity of the enzyme is extremely high: it acts only at the apurinic site in the sarcin/ricin domain of intact ribosomes, not on deproteinized rRNA or DNA containing apurinic sites, The amino acid sequences of five endopeptidase LysC-liberated peptides from the purified enzyme were determined and used to obtain a cDNA sequence. The open reading frame encodes a protein of 456 amino acids, and a homology search revealed a related rice protein. Similar enzyme activities were also found in other plants that express ribosome-inactivating proteins. We believe that RALyase is part of a complex self-defense mechanism.
引用
收藏
页码:6522 / 6531
页数:10
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