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Dynamic recruitment of single RNAs to processing bodies depends on RNA functionality
- Publication Year :
- 2018
- Publisher :
- Cold Spring Harbor Laboratory, 2018.
-
Abstract
- SUMMARYCellular RNAs often colocalize with cytoplasmic, membrane-less ribonucleoprotein (RNP) granules enriched for RNA processing enzymes, termed processing bodies (PBs). Here, we track the dynamic localization of individual miRNAs, mRNAs and long non-coding RNAs (lncRNAs) to PBs using intracellular single-molecule fluorescence microscopy. We find that unused miRNAs stably bind to PBs, whereas functional miRNAs, repressed mRNAs and lncRNAs both transiently and stably localize within either the core or periphery of PBs, albeit to different extents. Consequently, translation potential and positioning of cis-regulatory elements significantly impact PB-localization dynamics of mRNAs. Using computational modeling and supporting experimental approaches we show that phase separation into large PBs attenuates mRNA silencing, suggesting that physiological mRNA turnover predominantly occurs outside of PBs. Instead, our data support a role for PBs in sequestering unused miRNAs to regulate their surveillance and provides a framework for investigating the dynamic assembly of RNP granules by phase separation at single-molecule resolution.
- Subjects :
- RNA, Untranslated
Biology
Cytoplasmic Granules
03 medical and health sciences
0302 clinical medicine
microRNA
Fluorescence microscope
Humans
Gene silencing
Gene Silencing
RNA, Messenger
RNA Processing, Post-Transcriptional
Molecular Biology
030304 developmental biology
Ribonucleoprotein
0303 health sciences
Messenger RNA
Chemistry
030302 biochemistry & molecular biology
Colocalization
RNA
Translation (biology)
Cell Biology
Single Molecule Imaging
Cell biology
MicroRNAs
Ribonucleoproteins
Cytoplasm
RNA, Long Noncoding
030217 neurology & neurosurgery
HeLa Cells
Subjects
Details
- Language :
- English
- Database :
- OpenAIRE
- Accession number :
- edsair.doi.dedup.....78d1b5015c996a190fa1e2ffb4db71fa
- Full Text :
- https://doi.org/10.1101/375295