Evidence that siRNAs function as guides, not primers, in the Drosophila and human RNAi pathways
Graduate School of Biomedical Sciences; Department of Biochemistry and Molecular Pharmacology
Medical Subject Headings
Animals; Drosophila melanogaster; Endoribonucleases; *Gene Expression Regulation; Genes, Insect; Hela Cells; Humans; Phosphates; RNA Interference; RNA, Double-Stranded; RNA, Small Interfering; Ribonuclease III
Life Sciences | Medicine and Health Sciences
In Drosophila, two features of small interfering RNA (siRNA) structure--5' phosphates and 3' hydroxyls--are reported to be essential for RNA interference (RNAi). Here, we show that as in Drosophila, a 5' phosphate is required for siRNA function in human HeLa cells. In contrast, we find no evidence in flies or humans for a role in RNAi for the siRNA 3' hydroxyl group. Our in vitro data suggest that in both flies and mammals, each siRNA guides endonucleolytic cleavage of the target RNA at a single site. We conclude that the underlying mechanism of RNAi is conserved between flies and mammals and that RNA-dependent RNA polymerases are not required for RNAi in these organisms.
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Citation: Mol Cell. 2002 Sep;10(3):537-48.
Schwarz, Dianne S.; Hutvagner, Gyorgy; Haley, Benjamin; and Zamore, Phillip D., "Evidence that siRNAs function as guides, not primers, in the Drosophila and human RNAi pathways" (2002). GSBS Student Publications. 1085.