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| Status |
Public on Jan 20, 2021 |
| Title |
Antisense ribosomal siRNAs inhibit RNA polymerase I-directed transcription in C. elegans |
| Organism |
Caenorhabditis elegans |
| Experiment type |
Non-coding RNA profiling by high throughput sequencing
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| Summary |
Eukaryotic cells express a wide variety of endogenous small regulatory RNAs that function in the nucleus. We previously found that erroneous rRNAs induce the generation of antisense ribosomal siRNAs (risiRNAs) which silence the expression of rRNAs via the nuclear RNAi defective (Nrde) pathway. To further understand the biological roles and mechanisms of this class of small regulatory RNAs, we conducted forward genetic screening to identify factors involved in risiRNA generation in Caenorhabditis elegans. We found that risiRNAs accumulated in the RNA exosome mutants. risiRNAs directed a NRDE-dependent silencing of pre-rRNAs in the nucleolus. In the presence of risiRNAs, NRDE-2 accumulated in the nucleolus and colocalized with RNA polymerase I. risiRNAs inhibited the transcription elongation of RNA polymerase I by decreasing RNAP I occupancy downstream of the RNAi-targeted site. Meanwhile, exosomes mislocalized from the nucleolus to nucleoplasm in suppressor of siRNA (susi) mutants, in which erroneous rRNAs accumulated. These results established a novel model of rRNA surveillance by combining ribonuclease-mediated RNA degradation with small RNA-directed nucleolar RNAi system.
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| Overall design |
In this study, we compared total small RNAs from (1) wild type animal; (2) dis-3(ust56) mutant; (3) exos-1(ust57) mutant. And we compared total small RNAs from (4) eri-1(mg366) animal; (5) eri-1(mg366);dis-3(ust56) mutant; (6) eri-1(mg366);dis-3(ust56);rrf-1(pk1417);rrf-2(ok210) mutant;(7) eri-1(mg366);exos-1(ust57) mutant; (8) eri-1(mg366);exos-10(ok2269) mutant; And we compared the NRDE-3 associated small RNAs in (9) control animals; (10) dis-3(ust56) mutant; and (11) exos-1(ust57) mutant; (12) exos-10(ok2269); (13) dis-3(ust56);rrf-1(pk1417);rrf-2(ok210) mutant. And we also compared total small RNAs from (14) control animals; (15) fib-1 RNAi animals; (16) mtr-4 RNAi animals; (17) nol-56 RNAi animals;
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| Contributor(s) |
Guang S |
| Citation missing |
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| Submission date |
Jan 19, 2021 |
| Last update date |
Jan 23, 2021 |
| Contact name |
Shouhong Guang |
| E-mail(s) |
sguang@ustc.edu.cn
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| Organization name |
University of Science & Technology of China
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| Department |
School of Life Sciences
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| Street address |
443 HuangShan Road
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| City |
Hefei |
| State/province |
Anhui |
| ZIP/Postal code |
230027 |
| Country |
China |
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| Platforms (1) |
| GPL18245 |
Illumina HiSeq 2500 (Caenorhabditis elegans) |
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| Samples (17)
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| GSM5025700 |
Total_small_RNA_eri1(mg366) |
| GSM5025701 |
Total_small_RNA_eri1(mg366)_dis3(ust56) |
| GSM5025702 |
Total_small_RNA_eri1(mg366)_dis3(ust56)_rrf1(pk1417)_rrf2(ok210) |
| GSM5025704 |
Total_small_RNA_eri1(mg366)_exos1(ust57) |
| GSM5025705 |
Total_small_RNA_eri1(mg366)_exos10(ok2269) |
| GSM5025707 |
NRDE3_IP_small_RNA_eri1(mg366)_GFPNRDE3 |
| GSM5025708 |
NRDE3_IP_small_RNA_eri1(mg366)_dis3(ust56)_GFPNRDE3 |
| GSM5025709 |
NRDE3_IP_small_RNA_eri1(mg366)_exos1(ust57)_GFPNRDE3 |
| GSM5025711 |
NRDE3_IP_small_RNA_eri1(mg366)_exos10(ok2269)_GFPNRDE3 |
| GSM5025712 |
NRDE3_IP_small_RNA_eri1(mg366)_dis3(ust56)_rrf1(pk1417)_rrf2(ok210)_GFPNRDE3 |
| GSM5025714 |
Total_small_RNA_eri1(mg366)_L4440_RNAi |
| GSM5025715 |
Total_small_RNA_eri1(mg366)_fib1_RNAi |
| GSM5025717 |
Total_small_RNA_eri1(mg366)_mtr4_RNAi |
| GSM5025718 |
Total_small_RNA_eri1(mg366)_nol56_RNAi |
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| Relations |
| BioProject |
PRJNA693200 |
| SRA |
SRP302336 |