Transcriptomics

Dataset Information

4

Caenorhabditis elegans ALG-1 antimorphic mutations uncover functions for Argonaute in microRNA guide strand selection and passenger strand disposal


ABSTRACT: MicroRNAs are regulators of gene expression whose functions are critical for normal development and physiology. We have previously characterized mutations in a Caenorhabditis elegans microRNA-specific Argonaute ALG-1 (Argonaute-like gene) that are antimorphic [alg-1(anti)]. alg-1(anti) mutants have dramatically stronger microRNA-related phenotypes than animals with a complete loss of ALG-1. ALG-1(anti) miRISC (microRNA induced silencing complex) fails to undergo a functional transition from microRNA processing to target repression. To better understand this transition, we characterized the small RNA population associated with ALG-1(anti) complexes in vivo. alg-1(anti) mutants dramatically overaccumulated microRNA* (passenger) strands, and immunoprecipitated ALG-1(anti) complexes contained nonstoichiometric yields of mature microRNA and microRNA* strands, with some microRNA* strands present in the ALG-1(anti) Argonaute far in excess of the corresponding mature microRNAs. We show complex and microRNA-specific defects in microRNA strand selection and microRNA* strand disposal. For certain microRNAs (for example mir-58), microRNA guide strand selection by ALG-1(anti) appeared normal, but microRNA* strand release was inefficient. For other microRNAs (such as mir-2), both the microRNA and microRNA* strands were selected as guide by ALG-1(anti), indicating a defect in normal specificity of the strand choice. Our results suggest that wild-type ALG-1 complexes recognize structural features of particular microRNAs in the context of conducting the strand selection and microRNA* ejection steps of miRISC maturation. Deep-sequencing was performed on cDNA libraries made from total RNA and RNA immunoprecipitated with ALG-1 from mixed-staged populations of three strains: three biological replicates from wild-type animals and two biological replicates each from alg-1(ma192) and alg-1(ma202) mutant animals. In addition, deep-sequencing was performed on cDNA libraries made from L2-staged total RNA in two biological replicates from wildtype and alg-1(ma202) animals and one biological replicate of alg-1(ma192).

ORGANISM(S): Caenorhabditis elegans  

SUBMITTER: Victor R Ambros   Anna Y Zinovyeva  Isana Veksler-Lublinsky 

PROVIDER: E-GEOD-72659 | ArrayExpress | 2015-09-07

SECONDARY ACCESSION(S): GSE72659SRP063111PRJNA294604

REPOSITORIES: GEO, ArrayExpress, ENA

Dataset's files

Source:
Action DRS
E-GEOD-72659.idf.txt Idf
E-GEOD-72659.processed.1.zip Processed
E-GEOD-72659.processed.2.zip Processed
E-GEOD-72659.sdrf.txt Txt
Items per page:
1 - 4 of 4

Similar Datasets

2015-01-01 | S-EPMC4586838 | BioStudies
2016-01-01 | S-EPMC5147811 | BioStudies
2009-01-01 | S-EPMC2795325 | BioStudies
2015-09-01 | MSV000079287 | MassIVE
1000-01-01 | S-EPMC3866245 | BioStudies
2019-01-01 | S-EPMC6559381 | BioStudies
2009-11-28 | E-GEOD-19138 | ArrayExpress
2017-01-01 | S-EPMC5510005 | BioStudies
2013-01-01 | S-EPMC3820791 | BioStudies
2010-01-01 | S-EPMC2834287 | BioStudies