Cell stress is related to re-localization of Argonaute 2 and to decreased RNA interference in human cells
| Autor: | Georg Sczakiel, Christina Engel, Anke Detzer, Winfried Wünsche |
|---|---|
| Rok vydání: | 2010 |
| Předmět: |
Small interfering RNA
RNA-induced silencing complex Eukaryotic Initiation Factor-2 Oligonucleotides Argonaute Biology Cytoplasmic Granules Molecular biology Cell biology Cell Line RNA silencing Protein Transport Stress granule Lipofectamine RNA interference Stress Physiological Argonaute Proteins Genetics Humans RNA Interference Molecular Biology Intracellular |
| Zdroj: | Nucleic Acids Research |
| ISSN: | 1362-4962 |
| Popis: | Various kinds of stress on human cells induce the formation of endogenous stress granules (SGs). Human Argonaute 2 (hAgo2), the catalytic core component of the RNA-induced silencing complex (RISC), can be recruited to SGs as well as P-bodies (PBs) indicating that the dynamic intracellular distribution of hAgo2 in SGs, in PBs or at other sub-cellular sites could be related to the efficiency of the RNA interference (RNAi) machinery. Here, we studied the influence of heat shock, sodium arsenite (NaAsO2), cycloheximide (CHX) and LipofectamineTM 2000-mediated transfection of phosphorothioate (PS)-modified oligonucleotides (ON) on the intracellular localization of hAgo2 and the efficiency of RNAi. Fluorescence microscopy and sedimentation analysis of cell fractions indicate stress-induced accumulation of hAgo2 in SGs and the loss of distinctly composed complexes containing hAgo2 or their sub-cellular context. Transfection of cells with PS-ON induces cell stress that is phenotypically similar to the established inducers heat shock and NaAsO2. The intracellular re-distribution of hAgo2 is related to its increased metabolic stability and to decreased RNAi directed by microRNA or by short interfering RNA. Here, we propose a functional model of the relationship between cell stress, translocation of hAgo2 to SGs providing a depot function, and loss of RNAi activity. |
| Databáze: | OpenAIRE |
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