RRC ID 68936
Author Takahashi T, Zenno S, Ishibashi O, Takizawa T, Saigo K, Ui-Tei K.
Title Interactions between the non-seed region of siRNA and RNA-binding RLC/RISC proteins, Ago and TRBP, in mammalian cells.
Journal Nucleic Acids Res
Abstract Small interfering RNA (siRNA)-based RNA interference (RNAi) is widely used for target gene silencing in various organisms. We previously showed that 8-nt-long 5' proximal nucleotides, which include seed sequence (positions 2-8 from the 5' end of guide strand), and the complementary sequence of the passenger strand are capable of being simultaneously replaced with cognate deoxyribonucleotides without any substantial loss of gene silencing. In the present study, examination was made of RNA requirements in the non-seed region of siRNA. The non-seed region of siRNA was found to be subdivided into four domains, in which two nucleotide pairs (positions 13 and 14) were replaceable with cognate deoxyribonucleotides without reducing RNAi activity. However, RNA sequences at positions 9-12 and 15-18 were essential for effective gene silencing, and these two double-stranded RNA cores are required for binding of the trans-activation response RNA-binding protein (TRBP). The terminal RNA (positions 19-21) provided Argonaute protein binding sites. Argonaute binding was enhanced by the presence of RNAs at positions 15-18. Knockdown experiments showed that, unlike Argonaute and TRBP, Dicer was dispensable for RNAi. Based on these observations, we discuss possible RNA/protein and protein/protein interactions in RNA-induced silencing complex formation.
Volume 42(8)
Pages 5256-69
Published 2014-4-1
DOI 10.1093/nar/gku153
PII gku153
PMID 24561616
PMC PMC4005638
MeSH Animals Argonaute Proteins / metabolism* CHO Cells Cricetinae Cricetulus DNA / chemistry HeLa Cells Humans RNA Interference* RNA, Small Interfering / chemistry* RNA, Small Interfering / metabolism RNA-Binding Proteins / metabolism* Ribonuclease III / metabolism
IF 11.502
Human and Animal Cells CHO-K1