|
Status |
Public on Oct 02, 2009 |
Title |
High-throughput pyrosequencing of endogenous small RNAs from CSR-1 IP complexes and csr-1(tm892) and ego-1(om97) mutants |
Organism |
Caenorhabditis elegans |
Experiment type |
Non-coding RNA profiling by high throughput sequencing
|
Summary |
High-throughput pyrosequencing of endogenous small RNAs from CSR-1 IP complexes and csr-1(tm892) and ego-1(om97) mutants with corresponding controls. RNAi-related pathways regulate diverse processes, from developmental timing to transposon silencing. Here, we show that in C. elegans the Argonaute CSR-1, the RNA-dependent RNA polymerase EGO-1, the Dicer-related helicase DRH-3, and the Tudor-domain protein EKL-1 localize to chromosomes and are required for proper chromosome segregation. In the absence of these factors chromosomes fail to align at the metaphase plate and kinetochores do not orient to opposing spindle poles. Surprisingly, the CSR-1 interacting small RNAs (22G-RNAs) are antisense to thousands of germline-expressed protein-coding genes. Nematodes assemble holocentric chromosomes in which continuous kinetochores must span the expressed domains of the genome. We show that CSR-1 interacts with chromatin at target loci, but does not down-regulate target mRNA or protein levels. Instead, our findings support a model in which CSR-1 complexes target protein-coding domains to promote their proper organization within the holocentric chromosomes of C. elegans.
|
|
|
Overall design |
5 samples examined. Small RNAs that co-immunopercipitate with CSR-1 protein and input sample. Small RNAs from csr-1(tm892) and ego-1(om97) mutants and corresponding congenic wild type strain.
|
|
|
Contributor(s) |
Claycomb JM, Mello CC |
Citation(s) |
19804758 |
|
Submission date |
Sep 18, 2009 |
Last update date |
May 15, 2019 |
Contact name |
Julie M. Claycomb |
Organization name |
University of Massachusetts Medical School
|
Department |
Program in Molecular Medicine
|
Lab |
Craig Mello
|
Street address |
373 Plantation Street Biotech2 Suite 219
|
City |
Worcester |
State/province |
MA |
ZIP/Postal code |
01605 |
Country |
USA |
|
|
Platforms (1) |
GPL9269 |
Illumina Genome Analyzer II (Caenorhabditis elegans) |
|
Samples (5)
|
GSM454000 |
small RNAs that co-immunopercepitate with the argonaute CSR-1 |
GSM454001 |
small RNAs from wild type worms input control for the IP experiment |
GSM454002 |
small RNAs from csr-1 mutant animals |
GSM454003 |
small RNAs from ego-1 mutant animals |
GSM454004 |
small RNAs from wild type control for csr-1(tm892) and ego-1(om97) |
|
This SubSeries is part of SuperSeries: |
GSE18167 |
Gene expression in csr-1 mutants and sequencing of sRNAs from CSR-1 IP complexes and csr-1 and ego-1 mutants |
|
Relations |
SRA |
SRP001450 |
BioProject |
PRJNA123485 |