A specific Argonaute phosphorylation regulates the binding to microRNAs during C. elegans development.
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ABSTRACT:
PROVIDER: PRJNA814677 | ENA |
REPOSITORIES: ENA
Dataset's files
| Action | DRS | |||
|---|---|---|---|---|
| SRR18292078.fastq.gz | Fastqsanger.gz | |||
| SRR18292079.fastq.gz | Fastqsanger.gz | |||
| SRR18292080.fastq.gz | Fastqsanger.gz | |||
| SRR18292081.fastq.gz | Fastqsanger.gz | |||
| SRR18292082.fastq.gz | Fastqsanger.gz |
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Project description:Argonaute proteins are at the core of the microRNA-mediated gene silencing pathway essential for animals. In C. elegans, the microRNA-specific Argonautes ALG-1 and ALG-2 regulate multiple processes required for proper animal developmental timing and viability. Here, we identified a new phosphorylation site, serine 642, on ALG-1 that modulates microRNA association. Mutating ALG-1 serine 642 into a phospho-mimicking residue impairs microRNA binding and causes embryonic lethality and post-embryonic phenotypes that are common with alteration of microRNA functions. Monitoring microRNA levels in alg-1 phosphorylation mutant animals reveal that miRNA passenger strands strongly increase but are not preferentially loaded into ALG-1, indicating that the miRNA binding defects could also lead to miRNA duplexes accumulation. Our genetic and biochemical experiments support the protein kinase A KIN-1 as the putative kinase that phosphorylates ALG-1 serine 642. Altogether, our data indicate that PKA triggers the ALG-1 phosphorylation to regulate its microRNAs association during C. elegans development.
2022-09-10 | GSE198352 | GEO
Project description:Phosphorylation of the Argonaute ALG-1 regulates microRNA binding during C. elegans development
| PRJNA729562 | ENA
Project description:Argonaute proteins are at the core of the microRNA-mediated gene silencing pathway essential for animals. In C. elegans, the microRNA-specific Argonautes ALG-1 and ALG-2 regulate multiple processes required for proper animal developmental timing and viability. Here we identified a phosphorylation site on ALG-1 that modulates microRNA association. Mutating ALG-1 serine 642 into a phospho-mimicking residue impairs microRNA binding and causes embryonic lethality and post-embryonic phenotypes that are consistent with alteration of microRNA functions. Monitoring microRNA levels in alg-1 phosphorylation mutant animals shows that microRNA passenger strands increase in abundance but are not preferentially loaded into ALG-1, indicating that the miRNA binding defects could lead to microRNA duplex accumulation. Our genetic and biochemical experiments support protein kinase A (PKA) KIN-1 as the putative kinase that phosphorylates ALG-1 serine 642. Our data indicate that PKA triggers ALG-1 phosphorylation to regulate its microRNA association during C. elegans development.
| S-EPMC10436268 | biostudies-literature
Project description:Argonaute proteins associate with small non-coding RNAs such as microRNAs, piRNAs and siRNAs to regulate gene expression required for cell proliferation and differentiation. Thse Argonaute proteins are the major components of effector complexes of non-coding RNA pathways. In C. elegans, we identified PPM-2, a Mg2+/Mn2+-dependent serine/threonine phosphatase, as a new regulator of the small RNA pathways in C. elegans. We demonstrate that PPM-2 is involved in the regulation of the let-7 microRNA family but also in the regulation of nuclear RNAi as well as it contributes to the expression of piRNAs. The loss of ppm-2 affects the expression level of Type 1 piRNAs and the stability of ALG-1, PRG-1 and NRDE-3 Argonaute proteins by sending them to the proteasomal degradation pathway. Altogether, our findings suggest that PPM-2 is regulating the function of small non-coding RNA pathways by preventing Argonaute proteins to be degraded via the proteasomal pathway.
2021-08-01 | GSE136462 | GEO
Project description:Comprehensive discovery of endogenous Argonaute binding sites in C. elegans
2009-11-24 | GSE19138 | GEO
Project description:Here, we report a new phosphorylation site on ALG-1 that modulates its ability to bind miRNAs. Mutating ALG-1 S642 into a phosphomimetic residue strongly impairs binding to miRNAs. Furthermore, this mutation causes embryonic lethality which are not observed in animals depleted of alg-1 suggesting that it may consequently impair the normal function of its homolog alg-2. Quantification of miRNAs in the phosphorylation mutants of alg-1 reveals that the miRNA passenger strands are strongly increased but not preferentially loaded into ALG-1, indicating that the defects in miRNA binding may also lead to an accumulation of miRNA duplexes.
2022-12-19 | GSE174368 | GEO
Project description:MicroRNAs (miRNAs) perform critical functions in normal physiology and disease by associating with Argonaute proteins and downregulating partially complementary messenger RNAs (mRNAs). To identify new regulators of the miRNA pathway, we employed CRISPR-Cas9 genome-wide loss-of-function screening coupled with a fluorescent reporter of miRNA activity. Iterative rounds of screening revealed a novel mechanism whereby target engagement by Argonaute 2 (AGO2) triggers its hierarchical, multi-site phosphorylation by CSNK1A1 on a set of highly conserved residues (S824-S834), followed by rapid dephosphorylation by the ANKRD52-PPP6C phosphatase complex. Although genetic and biochemical studies demonstrated that AGO2 phosphorylation on these residues inhibits target mRNA binding, inactivation of this phosphorylation cycle globally impairs miRNA-mediated silencing. Analysis of the transcriptome-wide binding profile of non-phosphorylatable AGO2 revealed a dramatic expansion of the target repertoire bound at steady-state, effectively reducing the active pool of AGO2 on a per target basis. These findings support a model in which an AGO2 phosphorylation cycle stimulated by target engagement regulates miRNA:target interactions to maintain the global efficiency of miRNA-mediated silencing.
2017-01-17 | GSE89943 | GEO
Project description:MicroRNAs (miRNAs) perform critical functions in normal physiology and disease by associating with Argonaute proteins and downregulating partially complementary messenger RNAs (mRNAs). To identify new regulators of the miRNA pathway, we employed CRISPR-Cas9 genome-wide loss-of-function screening coupled with a fluorescent reporter of miRNA activity. Iterative rounds of screening revealed a novel mechanism whereby target engagement by Argonaute 2 (AGO2) triggers its hierarchical, multi-site phosphorylation by CSNK1A1 on a set of highly conserved residues (S824-S834), followed by rapid dephosphorylation by the ANKRD52-PPP6C phosphatase complex. Although genetic and biochemical studies demonstrated that AGO2 phosphorylation on these residues inhibits target mRNA binding, inactivation of this phosphorylation cycle globally impairs miRNA-mediated silencing. Analysis of the transcriptome-wide binding profile of non-phosphorylatable AGO2 revealed a dramatic expansion of the target repertoire bound at steady-state, effectively reducing the active pool of AGO2 on a per target basis. These findings support a model in which an AGO2 phosphorylation cycle stimulated by target engagement regulates miRNA:target interactions to maintain the global efficiency of miRNA-mediated silencing.
2017-01-17 | GSE89944 | GEO
Project description:MicroRNAs (miRNAs) perform critical functions in normal physiology and disease by associating with Argonaute proteins and downregulating partially complementary messenger RNAs (mRNAs). To identify new regulators of the miRNA pathway, we employed CRISPR-Cas9 genome-wide loss-of-function screening coupled with a fluorescent reporter of miRNA activity. Iterative rounds of screening revealed a novel mechanism whereby target engagement by Argonaute 2 (AGO2) triggers its hierarchical, multi-site phosphorylation by CSNK1A1 on a set of highly conserved residues (S824-S834), followed by rapid dephosphorylation by the ANKRD52-PPP6C phosphatase complex. Although genetic and biochemical studies demonstrated that AGO2 phosphorylation on these residues inhibits target mRNA binding, inactivation of this phosphorylation cycle globally impairs miRNA-mediated silencing. Analysis of the transcriptome-wide binding profile of non-phosphorylatable AGO2 revealed a dramatic expansion of the target repertoire bound at steady-state, effectively reducing the active pool of AGO2 on a per target basis. These findings support a model in which an AGO2 phosphorylation cycle stimulated by target engagement regulates miRNA:target interactions to maintain the global efficiency of miRNA-mediated silencing.
2017-01-17 | GSE89940 | GEO
Project description:Small RNAs associate with Argonaute proteins to control gene expression, impacting a wide range of cellular processes, such as antiviral defense, transposon silencing, and development. Plants and animals typically have several classes of small RNAs, along with multiple Argonautes. These Argonautes often confer distinct functionality to the various classes of small RNAs. But how small RNAs are selectively loaded into the appropriate Argonaute is not well understood. Small interfering RNAs (siRNAs) and microRNAs (miRNAs) are two classes of small RNAs commonly generated from double-stranded RNA (dsRNA) precursors by the endoribonuclease Dicer, although not all siRNAs or miRNAs are produced in this manner. dsRNA-derived siRNAs are often cleaved from fully base-paired precursors, whereas miRNAs typically originate from partially base-paired hairpins. In C. elegans, Dicer/DCR-1 processing of siRNAs and a related dsRNA-derived small RNA class known as 26G-RNAs is mediated by the dsRNA-binding protein RDE-4. In the study associated with this data, we used small RNA high-throughput sequencing to show that RDE-4 also promotes loading of siRNAs but not miRNAs into the Argonaute RDE-1, but not ALG-1, and loading of 26G-RNAs into the Argonaute ERGO-1, but not into the two paralogous Argonautes ALG-3 and ALG-4. Although ALG-3/4 associated 26G-RNAs do not require RDE-4 for Argonaute loading, the levels of these small RNAs are strongly reduced in rde-4 mutants. Therefore, RDE-4 functions in both dsRNA processing and selective loading of small RNAs into specific Argonautes. Our findings reveal a role for RDE-4 as a critical determinant of small RNA loading specificity and provide insight into the mechanisms by which small RNAs are selectively paired with their corresponding Argonaute proteins.
2025-04-08 | GSE293782 | GEO