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Nucleic Acids Research 2006 34(2):667-675; doi:10.1093/nar/gkj474
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Published online 30 January 2006

© The Author 2006. Published by Oxford University Press. All rights reserved
The online version of this article has been published under an open access model. Users are entitled to use, reproduce, disseminate, or display the open access version of this article for non-commercial purposes provided that: the original authorship is properly and fully attributed; the Journal and Oxford University Press are attributed as the original place of publication with the correct citation details given; if an article is subsequently reproduced or disseminated not in its entirety but only in part or as a derivative work this must be clearly indicated. For commercial re-use, please contact journals.permissions{at}oxfordjournals.org

Article

HEN1 recognizes 21–24 nt small RNA duplexes and deposits a methyl group onto the 2' OH of the 3' terminal nucleotide

Zhiyong Yang, Yon W. Ebright1, Bin Yu and Xuemei Chen*

Department of Botany and Plant Sciences, University of California Riverside, Riverside, CA 92521, USA 1Waksman Institute, Rutgers University Piscataway, NJ 08854, USA

*To whom correspondence should be addressed. Tel: +1 951 827 3988; Fax: +1 951 827 4437; Email: xuemei.chen{at}ucr.edu

Received December 15, 2005. Revised January 11, 2006. Accepted January 11, 2006.

microRNAs (miRNAs) and small interfering RNAs (siRNAs) in plants bear a methyl group on the ribose of the 3' terminal nucleotide. We showed previously that the methylation of miRNAs and siRNAs requires the protein HEN1 in vivo and that purified HEN1 protein methylates miRNA/miRNA* duplexes in vitro. In this study, we show that HEN1 methylates both miRNA/miRNA* and siRNA/siRNA* duplexes in vitro with a preference for 21–24 nt RNA duplexes with 2 nt overhangs. We also demonstrate that HEN1 deposits the methyl group on to the 2' OH of the 3' terminal nucleotide. Among various modifications that can occur on the ribose of the terminal nucleotide, such as 2'-deoxy, 3'-deoxy, 2'-O-methyl and 3'-O-methyl, only 2'-O-methyl on a small RNA inhibits the activity of yeast poly(A) polymerase (PAP). These findings indicate that HEN1 specifically methylates miRNAs and siRNAs and implicate the importance of the 2'-O-methyl group in the biology of RNA silencing.


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