Abstract
Expression of dsRNA complementary to small nucleolar RNAs (snoRNAs) in Trypanosoma brucei results in snoRNA silencing, termed snoRNAi. Here, we demonstrate that snoRNAi requires the nuclear TbDCL2 protein, but not TbDCL1, which is involved in RNA interference (RNAi) in the cytoplasm. snoRNAi depends on Argonaute1 (Slicer), and on TbDCL2, suggesting that snoRNA dicing and slicing takes place in the nucleus, and further suggesting that AGO1 is active in nuclear silencing. snoRNAi was next utilized to elucidate the function of an abundant snoRNA, TB11Cs2C2 (92nt), present in a cluster together with the spliced leader associated RNA (SLA1) and snR30, which are both H/ACA RNAs with special nuclear functions. Using AMT-UV cross-linking and RNaseH cleavage, we provide evidence for the interaction of TB11Cs2C2 with the small rRNAs, srRNA-2 and srRNA-6, which are part of the large subunit (LSU) rRNA. snoRNAi of TB11Cs2C2 resulted in defects in generating srRNA-2 and LSUβ rRNA. This is the first snoRNA described so far to engage in trypanosome-specific processing events.
| Original language | English |
|---|---|
| Pages (from-to) | 7236-7247 |
| Number of pages | 12 |
| Journal | Nucleic Acids Research |
| Volume | 38 |
| Issue number | 20 |
| DOIs | |
| State | Published - Nov 2010 |
Bibliographical note
Funding Information:Funding for open access charge: Israel-US Binational Science Foundation (BSF), by an International Research Scholar’s Grant from the Howard Hughes Foundation to SM; Public Health Service Grant (grant number AI28798) to EU. S.M. holds the David and Inez Myers Chair in RNA silencing of diseases.