Citation | Zhang L, Ding L, Cheung TH, Dong MQ, Chen J, Sewell AK, Liu X, Yates JR, Han M. Systematic identification of C. elegans miRISC proteins, miRNAs, and mRNA targets by their interactions with GW182 proteins AIN-1 and AIN-2. Mol Cell, 2007. |
PubMed ID | 18042455 |
Short Description | Systematic identification of C. elegans miRISC proteins, miRNAs, and mRNA targets by their interactions with GW182 proteins AIN-1 and AIN-2. GEO Record: GSE9073 Platform: GPL5883 Download gene-centric, log2 transformed data: WBPaper00031252.ce.mr.csv |
# of Conditions | 20 |
Full Description | MicroRNAs (miRNAs) regulate gene expression for diverse functions, but only a limited number of mRNA targets have been experimentally identified. We show that GW182 family proteins AIN-1 and AIN-2 act redundantly to regulate the expression of miRNA targets, but not miRNA biogenesis. Immunoprecipitation (IP) and mass spectrometry indicate that AIN-1 and AIN-2 interact only with miRNA-specific Argonaute proteins ALG-1 and ALG-2 and with components of the core translational initiation complex. Known miRNA targets are enriched in AIN-2 complexes, correlating with the expression of corresponding miRNAs. Combining IP with pyrosequencing and microarray analysis of RNAs associated with AIN-1/AIN-2, we identified 106 previously annotated miRNAs plus nine new candidate miRNAs, but nearly no siRNAs, and more than 3500 potential miRNA targets, including nearly all known ones. Our results demonstrate an effective biochemical approach to systematically identify miRNA targets and provide valuable insights regarding the properties of miRNA effector complexes. Experimental Details: WBPaper00031252:AIN-1_IP_rep1 WBPaper00031252:AIN-1_IP_rep2 WBPaper00031252:AIN-1_IP_rep3 WBPaper00031252:Preimmune_IP_rep1 WBPaper00031252:Preimmune_IP_rep2 WBPaper00031252:Preimmune_IP_rep3 WBPaper00031252:AIN-2-GFP_IP_rep1 WBPaper00031252:AIN-2-GFP_IP_rep2 WBPaper00031252:AIN-2-GFP_IP_rep3 WBPaper00031252:GFP_IP_rep1 WBPaper00031252:GFP_IP_rep2 WBPaper00031252:GFP_IP_rep3 WBPaper00031252:N2_totalRNA_rep1 WBPaper00031252:N2_totalRNA_rep2 WBPaper00031252:AIN-2-GFP_totalRNA_rep1 WBPaper00031252:AIN-2-GFP_totalRNA_rep2 WBPaper00031252:AIN-2-GFP_totalRNA_rep3 WBPaper00031252:AIN-2_promoter_GFP_totalRNA_rep1 WBPaper00031252:AIN-2_promoter_GFP_totalRNA_rep2 WBPaper00031252:AIN-2_promoter_GFP_totalRNA_rep3. |
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