A micrococcal nuclease homologue in RNAi effector complexes

Amy A. Caudy, René F. Ketting, Scott M. Hammond, Ahmet M. Denli, Anja M. P. Bathoorn, Bastiaan B. J. Tops, Jose M. Silva, Mike M. Myers, Gregory J. Hannon () and Ronald H. A. Plasterk ()
Additional contact information
Amy A. Caudy: Watson School of Biological Sciences
René F. Ketting: The Hubrecht Laboratory, Centre for Biomedical Genetics
Scott M. Hammond: Watson School of Biological Sciences
Ahmet M. Denli: Watson School of Biological Sciences
Anja M. P. Bathoorn: Watson School of Biological Sciences
Bastiaan B. J. Tops: Watson School of Biological Sciences
Jose M. Silva: Watson School of Biological Sciences
Mike M. Myers: Watson School of Biological Sciences
Gregory J. Hannon: Watson School of Biological Sciences
Ronald H. A. Plasterk: The Hubrecht Laboratory, Centre for Biomedical Genetics

Nature, 2003, vol. 425, issue 6956, 411-414

Abstract: Abstract RNA interference (RNAi) regulates gene expression by the cleavage of messenger RNA, by mRNA degradation and by preventing protein synthesis. These effects are mediated by a ribonucleoprotein complex known as RISC (RNA-induced silencing complex)1. We have previously identified four Drosophila components (short interfering RNAs1, Argonaute 2 (ref. 2), VIG and FXR3) of a RISC enzyme that degrades specific mRNAs in response to a double-stranded-RNA trigger. Here we show that Tudor-SN (tudor staphylococcal nuclease)—a protein containing five staphylococcal/micrococcal nuclease domains and a tudor domain—is a component of the RISC enzyme in Caenorhabditis elegans, Drosophila and mammals. Although Tudor-SN contains non-canonical active-site sequences, we show that purified Tudor-SN exhibits nuclease activity similar to that of other staphylococcal nucleases. Notably, both purified Tudor-SN and RISC are inhibited by a specific competitive inhibitor of micrococcal nuclease. Tudor-SN is the first RISC subunit to be identified that contains a recognizable nuclease domain, and could therefore contribute to the RNA degradation observed in RNAi.

Date: 2003
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DOI: 10.1038/nature01956

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