RDE-1 slicer activity is required only for passenger-strand cleavage during RNAi in Caenorhabditis elegans.
RNA interference (RNAi) is a process in which double-stranded RNA is cleaved into small interfering RNAs (siRNAs) that induce the destruction of homologous single-stranded mRNAs. Argonaute proteins are essential components of this silencing process; they bind siRNAs directly and can cleave RNA targets using a conserved RNase H motif. In Caenorhabditis elegans, the Argonaute protein RDE-1 has a central role in RNAi. In animals lacking RDE-1, the introduction of double-stranded RNA does not trigger any detectable level of RNAi. Here we show that RNase H activity of RDE-1 is required only for efficient removal of the passenger strand of the siRNA duplex and not for triggering the silencing response at the target-mRNA level. These results uncouple the role of the RDE-1 RNase H activity in small RNA maturation from its role in target-mRNA silencing in vivo.
Steiner
Florian A
FA
Hubrecht Institute, Royal Netherlands Academy of Arts and Sciences & University Medical Centre Utrecht, Uppsalalaan 8, 3584 CT, Utrecht, The Netherlands.
Okihara
Kristy L
KL
Hoogstrate
Suzanne W
SW
Sijen
Titia
T
Ketting
René F
RF
D000818:Animals; D017173:Caenorhabditis elegans / Q000378:metabolism*; D029742:Caenorhabditis elegans Proteins / Q000378:metabolism*; D034622:RNA Interference; D020871:RNA Stability; D012330:RNA, Double-Stranded / Q000378:metabolism; D034741:RNA, Small Interfering / Q000378:metabolism; D016914:Ribonuclease H / Q000378:metabolism
D029742:Caenorhabditis elegans Proteins; D012330:RNA, Double-Stranded; D034741:RNA, Small Interfering; C122185:rde-1 protein, C elegans; D016914:Ribonuclease H