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J. Virol. doi:10.1128/JVI.00885-07
Copyright (c) 2007, American Society for Microbiology and/or the Listed Authors/Institutions. All Rights Reserved.

Adenovirus VA RNAII derived small RNAs are efficiently incorporated into RISC and associate with polyribosomes

Department of Medical Biochemistry and Microbiology, Uppsala Biomedical Center, Husargatan 3, S-751 23 Uppsala, Sweden

^* To whom correspondence should be addressed. Email: goran.akusjarvi{at}imbim.uu.se.

	Abstract

Adenovirus type 5 encodes two highly structured short RNAs, the virus-associated RNAs, VA RNAI and VA RNAII. Both are processed by Dicer into small RNAs that are incorporated into the RNA-induced silencing complex (RISC). We here show, by cloning of small RNAs, that approximately 80% of Ago2-containing RISC immunopurified from late infected cells, is associated with VA RNA-derived small RNAs (mivaRNAs). Most surprisingly, VA RNAII, which is expressed at 20-fold lower levels compared to VA RNAI, appears to be the preferred substrate for Dicer and accounts for approximately 60% of all small RNAs in RISC. The mivaRNAs are derived from the 3' strand of the terminal stems of the VA RNAs, with the major fraction of VA RNAII starting at position 138. The small RNAs derived from VA RNAI were more heterogeneous in size with the two predominant small RNAs starting at position 137 and 138. Collectively, our results suggest that the mivaRNAs are efficiently used for RISC assembly in late infected cells. Potentially they function as miRNAs regulating translation of cellular mRNAs. In support of this hypothesis, we detect a fraction of the VA RNAII derived mivaRNAs on polyribosomes