Nuclear localization of Argonaute 2 is affected by cell density and may relieve repression by microRNAs Journal Article
| Authors: | Johnson, K. C.; Kilikevicius, A.; Hofman, C.; Hu, J.; Liu, Y.; Aguilar, S.; Graswich, J.; Han, Y.; Wang, T.; Westcott, J. M.; Brekken, R. A.; Peng, L.; Karagkounis, G.; Corey, D. R. |
| Article Title: | Nuclear localization of Argonaute 2 is affected by cell density and may relieve repression by microRNAs |
| Abstract: | Argonaute protein is associated with post-transcriptional control of cytoplasmic gene expression through miRNA-induced silencing complexes (miRISC). Specific cellular and environmental conditions can trigger AGO protein to accumulate in the nucleus. Localization of AGO is central to understanding miRNA action, yet the consequences of AGO being in the nucleus are undefined. We show nuclear enrichment of AGO2 in HCT116 cells grown in two-dimensional culture to high density, HCT116 cells grown in three-dimensional tumor spheroid culture, and human colon tumors. The shift in localization of AGO2 from cytoplasm to nucleus de-represses cytoplasmic AGO2-eCLIP targets that were candidates for canonical regulation by miRISC. Constitutive nuclear localization of AGO2 using an engineered nuclear localization signal increases cell migration. Critical RNAi factors also affect the localization of AGO2. Knocking out an enzyme essential for miRNA biogenesis, DROSHA, depletes mature miRNAs and restricts AGO2 localization to the cytoplasm, while knocking out the miRISC scaffolding protein, TNRC6, results in nuclear localization of AGO2. These data suggest that AGO2 localization and miRNA activity can be regulated depending on environmental conditions, expression of mature miRNAs, and expression of miRISC cofactors. Localization and expression of core miRISC protein machinery should be considered when investigating the roles of miRNAs. © The Author(s) 2023. |
| Keywords: | controlled study; human cell; genetics; flow cytometry; metabolism; cell viability; reverse transcription polymerase chain reaction; apoptosis; microrna; gene expression; confocal microscopy; rna interference; transcriptomics; fluorescence in situ hybridization; amino acid sequence; colon tumor; western blotting; immunoprecipitation; dactinomycin; cell migration; cytoplasm; cell count; real time polymerase chain reaction; cell density; micrornas; nuclear localization signal; argonaute protein; fluorescence activated cell sorting; transcriptome; rna splicing; student t test; rna extraction; argonaute 1 protein; biogenesis; argonaute 2 protein; rna sequence; stoichiometry; arabidopsis thaliana; argonaute proteins; tumor spheroid; humans; human; article; hela cell line; defense mechanism; coimmunoprecipitation; crispr-cas9 system; hct 116 cell line; fibroblast cell line |
| Journal Title: | Nucleic Acids Research |
| Volume: | 52 |
| Issue: | 4 |
| ISSN: | 0305-1048 |
| Publisher: | Oxford University Press |
| Date Published: | 2024-02-25 |
| Start Page: | 1930 |
| End Page: | 1952 |
| Language: | English |
| DOI: | 10.1093/nar/gkad1155 |
| PUBMED: | 38109320 |
| PROVIDER: | scopus |
| PMCID: | PMC10899759 |
| DOI/URL: | |
| Notes: | Source: Scopus |
