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Series GSE201732 Query DataSets for GSE201732
Status Public on May 05, 2023
Title Circadian ribosome profiling reveals a role for the Period2 upstream opening reading frame in sleep
Organism Mus musculus
Experiment type Other
Summary Many mammalian proteins have circadian cycles of production and degradation, but de novo transcription is only responsible for a small fraction of this rhythmicity. We used ribosome profiling to quantify RNA translation in liver from circadian-entrained mice transferred to constant darkness conditions over a 24-h period and compared translation levels to absolute protein production for 16 circadian proteins. We observed a delay between translation and peak protein levels for several circadian genes covering a wide range of translation efficiencies. We found extensive binding of ribosomes to upstream open reading frames (uORFs) in circadian mRNAs, including the core clock gene Period2 (Per2). Increased uORFs in 5' UTRs was associated with decreased ribosome binding in downstream ORFs and reduced expression of synthetic reporter constructs in vitro and in single cells. Mutation of the Per2 uORF increased luciferase and fluorescence reporter expression in 3T3 cells without altering phase or period. Genomic Per2 uORF mutation using CRISPR/Cas9 increased PER2 expression in PER2:Luc MEFs and reduced sleep in Per2 uORF mutant mice. These results suggest that post-transcriptional processes shape translation of mRNA transcripts, which can impact physiological rhythms and sleep.
 
Overall design Circadian ribosome profiling of two C57BL/6N mouse livers from each timepoint CT00, CT04, CT08, CT12, CT16, CT20, and CT24
Web link https://www.pnas.org/doi/10.1073/pnas.2214636120
 
Contributor(s) Millius A, Perrin D, Ueda HR
Citation(s) 37769257
Submission date Apr 27, 2022
Last update date Dec 13, 2023
Contact name Arthur Millius
E-mail(s) arthur.millius@ifrec.osaka-u.ac.jp
Phone +81-06-6879-8366
Organization name Osaka University
Department iFReC
Lab Systems Immunology
Street address iFReC 6F
City Suita
State/province Osaka
ZIP/Postal code 565-0871
Country Japan
 
Platforms (1)
GPL18480 Illumina HiSeq 1500 (Mus musculus)
Samples (15)
GSM6070353 A1_CT00_130516liver_8am (CT00_rep1)
GSM6070354 A2_CT04_130516liver_12pm (CT04_rep1)
GSM6070355 A3_CT08_130516liver_4pm (CT08_rep1)
Relations
BioProject PRJNA832792

Download family Format
SOFT formatted family file(s) SOFTHelp
MINiML formatted family file(s) MINiMLHelp
Series Matrix File(s) TXTHelp

Supplementary file Size Download File type/resource
GSE201732_RAW.tar 4.2 Mb (http)(custom) TAR (of TXT)
GSE201732_Supplementary_Table_2_JTK_RiboProf_v4metaAlloc_.xlsx 4.5 Mb (ftp)(http) XLSX
SRA Run SelectorHelp
Raw data are available in SRA
Processed data provided as supplementary file
Processed data are available on Series record

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