|
| Status |
Public on Dec 19, 2016 |
| Title |
Distinct Functions of Argonaute Slicer in siRNA Maturation and Heterochromatin Formation |
| Organism |
Schizosaccharomyces pombe |
| Experiment type |
Genome binding/occupancy profiling by high throughput sequencing
Non-coding RNA profiling by high throughput sequencing
|
| Summary |
Small-RNA (sRNA)-guided transcriptional gene silencing by Argonaute (Ago)-containing complexes is fundamental to genome integrity and epigenetic inheritance. The RNA cleavage (“Slicer”) activity of Argonaute has been implicated in both sRNA maturation and target RNA cleavage. Typically, Argonaute slices and releases the passenger strand of duplex sRNA to generate active silencing complexes, but it remains unclear whether slicing of target nascent RNAs, or other RNAi components, also contributes to downstream transcriptional silencing. Here, we develop a strategy for loading the fission yeast Ago1 with a single-stranded sRNA guide, which bypasses the requirement for slicer activity in generation of active silencing complexes. We show that slicer-defective Ago1 can mediate secondary sRNA generation, H3K9 methylation, and silencing similar to or better than wild-type and associates with chromatin more efficiently. The results define an ancient and minimal sRNA-mediated chromatin silencing mechanism, which resembles the germline-specific sRNA-dependent transcriptional silencing pathways in Drosophila and mammals.
This SuperSeries is composed of the SubSeries listed below.
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| |
|
| Overall design |
Refer to individual Series
|
| |
|
| Citation(s) |
27397687 |
| NIH grant(s) |
| Grant ID |
Grant title |
Affiliation |
Name |
| R01 GM072805 |
Epigenetic Inheritance of Heterochromatin |
HARVARD UNIVERSITY (MEDICAL SCHOOL) |
DANESH MOAZED |
|
| |
| Submission date |
May 23, 2016 |
| Last update date |
May 15, 2019 |
| Contact name |
Nahid Iglesias |
| E-mail(s) |
nahid.iglesias@duke.edu
|
| Phone |
919-613-3385
|
| Organization name |
Duke University
|
| Department |
Biomedical Engineering
|
| Lab |
Charles Gersbach
|
| Street address |
101 Science Drive, CIEMAS Rm. 2323
|
| City |
Durham |
| State/province |
NC |
| ZIP/Postal code |
27708 |
| Country |
USA |
| |
|
| Platforms (1) |
| GPL13988 |
Illumina HiSeq 2000 (Schizosaccharomyces pombe) |
|
| Samples (10)
|
| GSM2174859 |
ura4+::KAN + OE Ago1 WT (spy3442 + pDM817) |
| GSM2174860 |
ura4+::priRNA1-KAN ago1∆ + empty vector (spy3473 + pDM829) |
| GSM2174861 |
ura4+::priRNA1-KAN ago1∆ + OE Ago1 WT (spy3473 + pDM817) |
| GSM2174862 |
ura4+::priRNA1-KAN ago1∆ + OE Ago1 D580A (spy3473+ pDM831) |
| GSM2174863 |
ura4+::KAN leo1∆ + empty vector (spy6421 + pDM829) |
| GSM2174864 |
ura4+::priRNA1-KAN leo1∆ + empty vector (spy6419 + pDM829) |
| GSM2174865 |
Untagged WT (spy137) |
| GSM2174866 |
3xFlag-Ago1 WT (spy797) |
| GSM2174867 |
ura4+::priRNA1 ago1∆ + OE Flag-Ago1 WT (spy3473 + pDM817) - sRNA sense |
| GSM2174868 |
ura4+::priRNA1 ago1∆ + OE Flag-Ago1 D580A (spy3473 + pDM831) - sRNA sense |
|
| This SuperSeries is composed of the following SubSeries: |
| GSE81731 |
Distinct Functions of Argonaute Slicer in siRNA Maturation and Heterochromatin Formation [ChIP-Seq] |
| GSE81732 |
Distinct Functions of Argonaute Slicer in siRNA Maturation and Heterochromatin Formation [ncRNA-seq 1] |
| GSE81733 |
Distinct Functions of Argonaute Slicer in siRNA Maturation and Heterochromatin Formation [ncRNA-seq 2] |
|
| Relations |
| BioProject |
PRJNA322453 |
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