Abstract
Caenorhabditis elegans contains 25 Argonautes, of which, ALG-1 and ALG-2 are known to primarily interact with miRNAs. ALG-5 belongs to the AGO subfamily of Argonautes that includes ALG-1 and ALG-2, but its role in small RNA pathways is unknown. We analyzed by high-throughput sequencing the small RNAs associated with ALG-5, ALG-1 and ALG-2, as well as changes in mRNA expression in alg-5, alg-1 and alg-2 mutants. We show that ALG-5 defines a distinct branch of the miRNA pathway affecting the expression of genes involved in immunity, defense, and development. In contrast to ALG-1 and ALG-2, which associate with most miRNAs and have general roles throughout development, ALG-5 interacts with only a small subset of miRNAs and is specifically expressed in the germline where it localizes alongside the piRNA and siRNA machinery at P granules. alg-5 is required for optimal fertility and mutations in alg-5 lead to a precocious transition from spermatogenesis to oogenesis. Our results provide a near-comprehensive analysis of miRNA-Argonaute interactions in C. elegans and reveal a new role for miRNAs in the germline.
© The Author(s) 2017. Published by Oxford University Press on behalf of Nucleic Acids Research.
MeSH terms
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Animals
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Argonaute Proteins / genetics*
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Argonaute Proteins / metabolism
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Caenorhabditis elegans / classification
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Caenorhabditis elegans / genetics*
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Caenorhabditis elegans / growth & development
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Caenorhabditis elegans / metabolism
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Caenorhabditis elegans Proteins / genetics*
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Caenorhabditis elegans Proteins / metabolism
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Gene Expression Regulation, Developmental*
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Germ Cells / growth & development
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Germ Cells / metabolism*
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Hermaphroditic Organisms / genetics
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Hermaphroditic Organisms / growth & development
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Hermaphroditic Organisms / metabolism
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High-Throughput Nucleotide Sequencing
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MicroRNAs / genetics
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MicroRNAs / metabolism
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Mutation
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Oogenesis / genetics
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Phylogeny
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Protein Isoforms / genetics
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Protein Isoforms / metabolism
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RNA, Helminth / genetics*
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RNA, Helminth / metabolism
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RNA, Messenger / genetics
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RNA, Messenger / metabolism
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RNA, Small Interfering / genetics
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RNA, Small Interfering / metabolism
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RNA-Binding Proteins / genetics*
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RNA-Binding Proteins / metabolism
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Spermatogenesis / genetics
Substances
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ALG-1 protein, C elegans
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ALG-2 protein, C elegans
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ALG-5 protein, C elegans
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Argonaute Proteins
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Caenorhabditis elegans Proteins
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MicroRNAs
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Protein Isoforms
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RNA, Helminth
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RNA, Messenger
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RNA, Small Interfering
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RNA-Binding Proteins