UMMS Affiliation

Program in Molecular Medicine

Publication Date

2014-04-24

Document Type

Article

Subjects

Alleles; Amino Acid Sequence; Animals; Argonaute Proteins; Caenorhabditis elegans; Caenorhabditis elegans Proteins; Carrier Proteins; Conserved Sequence; Gene Expression Regulation, Developmental; MicroRNAs; Molecular Sequence Data; Mutation; Repressor Proteins; Sequence Alignment

Disciplines

Biochemistry | Developmental Biology | Molecular Biology | Molecular Genetics

Abstract

microRNAs function in diverse developmental and physiological processes by regulating target gene expression at the post-transcriptional level. ALG-1 is one of two Caenorhabditis elegans Argonautes (ALG-1 and ALG-2) that together are essential for microRNA biogenesis and function. Here, we report the identification of novel antimorphic (anti) alleles of ALG-1 as suppressors of lin-28(lf) precocious developmental phenotypes. The alg-1(anti) mutations broadly impair the function of many microRNAs and cause dosage-dependent phenotypes that are more severe than the complete loss of ALG-1. ALG-1(anti) mutant proteins are competent for promoting Dicer cleavage of microRNA precursors and for associating with and stabilizing microRNAs. However, our results suggest that ALG-1(anti) proteins may sequester microRNAs in immature and functionally deficient microRNA Induced Silencing Complexes (miRISCs), and hence compete with ALG-2 for access to functional microRNAs. Immunoprecipitation experiments show that ALG-1(anti) proteins display an increased association with Dicer and a decreased association with AIN-1/GW182. These findings suggest that alg-1(anti) mutations impair the ability of ALG-1 miRISC to execute a transition from Dicer-associated microRNA processing to AIN-1/GW182 associated effector function, and indicate an active role for ALG/Argonaute in mediating this transition.

Keywords

MicroRNAs, Phenotypes, Alleles, Mutation, RNA extraction, Immunoprecipitation, Larvae, Biosynthesis

Rights and Permissions

Copyright: © 2014 Zinovyeva et al. This is an open-access article distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited.

DOI of Published Version

10.1371/journal.pgen.1004286

Source

PLoS Genet. 2014 Apr 24;10(4):e1004286. doi: 10.1371/journal.pgen.1004286. eCollection 2014. Link to article on publisher's site

Journal/Book/Conference Title

PLoS genetics

Related Resources

Link to Article in PubMed

PubMed ID

24763381

Creative Commons License

Creative Commons Attribution 4.0 License
This work is licensed under a Creative Commons Attribution 4.0 License.

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