Minimal upstream open reading frame of Per2 mediates phase fitness of the circadian clock to day/night physiological body temperature rhythm

Miyake, Takahito; Inoue, Yuichi; Shao, Xinyan; Seta, Takehito; Aoki, Yuriko; Pham, Khanh Tien Nguyen; Shichino, Yuichi; Sasaki, Junko; Sasaki, Takehiko; Ikawa, Masahito; Yamaguchi, Yoshiaki; Okamura, Hitoshi; Iwasaki, Shigeo; Doi, Masao

doi:10.1016/j.celrep.2023.112157

Cited by 12 publications

(19 citation statements)

References 62 publications

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“…However, Per2 mRNA expression does not increase upon knockdown of the NMD component Smg6 ( 61 ), which suggests that other proteins in NMD independent of SMG6 may be involved ( 62 ). Another group deleted the start codon of the Per2 uORF but found no effect on Per2 mRNA levels and did not perform ribosome profiling ( 63 ), so it is unclear whether ribosome binding in Per2 is elevated in their Per2 uORF mutant mice as it is in our mutant mice. In both studies, PER2 protein levels in the mutant mice were similar to that in wild-type mice, indicating that circadian proteostasis pathways compensate for the increase in PER2 protein production.…”

Section: Discussionmentioning

confidence: 89%

Circadian ribosome profiling reveals a role for the Period2 upstream open reading frame in sleep

Millius,

Yamada,

Fujishima

et al. 2023

Proc. Natl. Acad. Sci. U.S.A.

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Many mammalian proteins have circadian cycles of production and degradation, and many of these rhythms are altered posttranscriptionally. We used ribosome profiling to examine posttranscriptional control of circadian rhythms by quantifying RNA translation in the liver over a 24-h period from circadian-entrained mice transferred to constant darkness conditions and by comparing ribosome binding levels to protein levels for 16 circadian proteins. We observed large differences in ribosome binding levels compared to protein levels, and we observed delays between peak ribosome binding and peak protein abundance. We found extensive binding of ribosomes to upstream open reading frames (uORFs) in circadian mRNAs, including the core clock gene Period2 (Per2) . An increase in the number of uORFs in the 5′UTR was associated with a decrease in ribosome binding in the main coding sequence and a reduction in expression of synthetic reporter constructs. Mutation of the Per2 uORF increased luciferase and fluorescence reporter expression in 3T3 cells and increased luciferase expression in PER2:LUC MEF cells. Mutation of the Per2 uORF in mice increased Per2 mRNA expression, enhanced ribosome binding on Per2 , and reduced total sleep time compared to that in wild-type mice. These results suggest that uORFs affect mRNA posttranscriptionally, which can impact physiological rhythms and sleep.

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Section: Discussionmentioning

confidence: 89%

Circadian ribosome profiling reveals a role for the Period2 upstream open reading frame in sleep

Millius,

Yamada,

Fujishima

et al. 2023

Proc. Natl. Acad. Sci. U.S.A.

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“…[1][2][3][4] Recently, we reported the identification of a translation-based mechanism of temperature-dependent circadian clock entrainment. 5) Particularly, using mouse embryonic fibroblast cells (MEFs) as a model system of peripheral tissue clock, we demonstrated that a physiological warm temperature shift, which we call warm temperature shift (WTS), within the physiologic range (a shift from 35 to 38.5 °C in culture) 5) has the ability to increase protein synthesis of the core clock gene Per2. This WTS-dependent upregulation of Per2 protein expression does not require transcriptional de novo synthesis of Per2 mRNA but it does require upregulation of Per2 translation through its minimal upstream open reading frame (m-uORF) located in the 5′ UTR of Per2.…”

Section: Introductionmentioning

confidence: 99%

“…This WTS-dependent upregulation of Per2 protein expression does not require transcriptional de novo synthesis of Per2 mRNA but it does require upregulation of Per2 translation through its minimal upstream open reading frame (m-uORF) located in the 5′ UTR of Per2. 5) Previously, we found that WTS can cause upregulation of phosphatidylinositol 3-kinase (PI3K) activity and reported its involvement in mediating WTS-dependent Per2 protein expression. 5) However, signaling pathway mediating the Per2 protein expression in response to WTS is thus far only sparsely understood.…”

Section: Introductionmentioning

confidence: 99%

Temperature-Dependent Upregulation of Per2 Protein Expression Is Mediated by eIF2α Kinases PERK and PKR through PI3K Activation

Shao,

Miyake,

Inoue

et al. 2024

Biological & Pharmaceutical Bulletin

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Temperature-dependent translational control of the core clock gene Per2 plays an important role in establishing entrainment of the circadian clock to physiological body temperature cycles. Previously, we found an involvement of the phosphatidylinositol 3-kinase (PI3K) in causing Per2 protein expression in response to a warm temperature shift (WTS) within a physiological range (from 35 to 38.5 °C). However, signaling pathway mediating the Per2 protein expression in response to WTS is only sparsely understood. Additional factor(s) other than PI3K remains unknown. Here we report the identification of eukaryotic initiation factor 2α (eIF2α) kinases, protein kinase R (PKR) and PKR-like endoplasmic reticulum kinase (PERK), as a novel mediator of WTS-dependent Per2 protein expression. Canonically, eIF2α has been regarded as a major downstream target of PERK and PKR. However, we found that PERK and PKR mediate WTS response of Per2 in a manner not involving eIF2α. We observed that PERK and PKR serve as an upstream regulator of PI3K rather than eIF2α in the context of WTS-dependent Per2 protein expression. There have been studies reporting PI3K activation occurring depending on PERK and PKR, while its physiological contribution has remained elusive. Our finding therefore not only helps to enrich the knowledge of how WTS affects Per2 protein expression but also extends the region of cellular biology involving the PERK/PKR-mediated PI3K activation to include entrainment-mechanism of the circadian clock.

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“…Although Neurospora and Arabidopsis share similar design principles in circadian rhythms as humans and flies, the proteins involved differ significantly in both structure and conservation [35][36][37] . Recent studies have shown that uORFs in the mouse core clock gene Period2 (Per2) play a role in temperature entrainment of the circadian clock 38 and in regulating sleep 39 . However, no significant differences in the period and amplitude of circadian rhythms were observed between wild-type and Per2 uORF mutant mice 39 .…”

Section: Introductionmentioning

confidence: 99%

“…By generating transgenic plants expressing core clock gene TIMING OF CAB EXPRESSION 1 with WT or mutant uORFs, Wu and colleagues identified a role for these uORFs in contributing to the robust operation of the circadian clock ( 18 ). Two other groups reported uORFs in the mouse core clock gene Period2 to be involved in temperature entrainment of the circadian clock ( 19 ) and regulating sleep ( 20 ). While the regulatory potential of uORFs in circadian rhythm is established, a detailed molecular delineation remains a coveted objective.…”

Section: Introductionmentioning

confidence: 99%

Upstream open reading frames dynamically modulate CLOCK protein translation to regulate circadian rhythm and sleep

Sun,

Shui,

et al. 2023

Preprint

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SummaryThe circadian rhythm is an evolutionarily conserved mechanism with translational regulation increasingly recognized as pivotal in its modulation. In this study, we show evidence that the upstream open reading frames (uORFs) of the core circadian clock gene,Clock(Clk), rhythmically and substantially attenuate CLK protein translation inDrosophila, with pronounced suppression occurring during daylight hours. EliminatingClkuORFs results in elevated CLK protein levels during the day and a compressed circadian cycle, along with a broad shift in clock gene expression rhythms. Interestingly,ClkuORF deletion also augments morning sleep by reducing dopaminergic activity. Beyond daily circadian adjustments,ClkuORFs play a role in modulating sleep patterns in response to the varying day lengths of different seasons, inhibiting translation in a day-length contingent manner. Furthermore, theClkuORFs act as a master regulator to shape the rhythmic expression of a vast array of genes and influencing multifaceted physiological outcomes. Collectively, our research sheds light on the intricate ways uORFs dynamically adjust downstream coding sequences to acclimate to environmental shifts.

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Minimal upstream open reading frame of Per2 mediates phase fitness of the circadian clock to day/night physiological body temperature rhythm

Cited by 12 publications

References 62 publications

Circadian ribosome profiling reveals a role for the Period2 upstream open reading frame in sleep

Circadian ribosome profiling reveals a role for the Period2 upstream open reading frame in sleep

Temperature-Dependent Upregulation of Per2 Protein Expression Is Mediated by eIF2α Kinases PERK and PKR through PI3K Activation

Upstream open reading frames dynamically modulate CLOCK protein translation to regulate circadian rhythm and sleep

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