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10:34
Detection of RNA-binding Proteins by In Vitro RNA Pull-down in Adipocyte Culture
Published on: July 22, 2016
Cold-induced RNA-binding proteins regulate circadian gene expression by controlling alternative polyadenylation
Yuting Liu1, Wenchao Hu, Yasuhiro Murakawa
1CAS-MPG Partner Institute for Computational Biology, Shanghai Institutes of Biological Sciences, Chinese Academy of Sciences, Shanghai, 200031, China.
Scientific Reports
|June 25, 2013
Summary
Two cold-induced RNA-binding proteins, Cirbp and Rbm3, control circadian gene expression by regulating alternative polyadenylation. Their depletion disrupts circadian rhythms by altering 3'UTR length and polyadenylation site selection.
Area of Science:
- Chronobiology
- Molecular Biology
- Gene Regulation
Background:
- Body temperature synchronizes mammalian circadian rhythms via unknown mechanisms.
- Circadian gene expression is crucial for physiological processes.
- RNA-binding proteins (RBPs) play roles in post-transcriptional regulation.
Purpose of the Study:
- To investigate the role of cold-induced RBPs, Cirbp and Rbm3, in temperature-entrained circadian gene expression.
- To elucidate the mechanism by which these RBPs regulate circadian rhythms.
Main Methods:
- Depletion of Cirbp and Rbm3 in mammalian cells.
- Poly(A)-tail-specific RNA immunoprecipitation followed by sequencing (PAR-CLIP) to identify RBP binding sites.
- Analysis of 3'UTR length and polyadenylation site (PAS) usage.
- Measurement of core circadian gene expression amplitudes.
Main Results:
- Depletion of Cirbp or Rbm3 significantly reduced the amplitude of core circadian genes.
- Cirbp and Rbm3 bind to 3'UTRs near polyadenylation sites.
- Cirbp/Rbm3 depletion shortened 3'UTRs; low temperature (upregulating Cirbp/Rbm3) lengthened them.
- Cirbp and Rbm3 repress proximal PAS usage, influencing circadian oscillations in PAS selection.
Conclusions:
- Cirbp and Rbm3 are key regulators of temperature-entrained circadian gene expression.
- These RBPs control circadian rhythms by modulating alternative polyadenylation (APA).
- APA is a critical mechanism for synchronizing peripheral clocks to body temperature cues.
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