Non-transcriptional processes in circadian rhythm generation

Wong, David; O’Neill, John S.

doi:10.1016/j.cophys.2018.10.003

Cited by 39 publications

(57 citation statements)

References 241 publications

(307 reference statements)

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“…These results show that cell cycle arrest at any stage suppresses the 4 hr rhythm. Therefore, 15 even though the dry mass oscillations we observed were not a direct consequence of the cell cycle, the oscillations seem to be a feature of proliferating cells.…”

Section: Role Of Cell Cycle and Circadian Oscillatorsmentioning

confidence: 60%

“…S7A). Despite the variability in cell cycle time, ranging in our experiments between 12.1h 15 and 21.3h, all analyzed cell lines/types showed the same oscillation period of 4 hours ( Fig. 1E), suggesting that the rhythm was not a cell-cycle harmonic.…”

Section: Role Of Cell Cycle and Circadian Oscillatorsmentioning

confidence: 67%

“…1A. We used lens-free microscopy to 15 analyze a large number of cells over several days by taking holographic images every 5 minutes.…”

Section: The 4 Hr Rhythmmentioning

confidence: 99%

“…1B). Excluding the 1/f noise, reconstruction of the raw signal by applying an inverse Fourier transform suggests that the cell dry mass shows periodic symmetric 15 Using this procedure, we studied several cell lines: Hela, U2OS, MEF, CHO-K1, NHEK and NHDF cells, which were immortalized from donors. In all of the cells, we observed the same principal frequency of 0.004 min -1 with the higher harmonics (Fig.…”

Section: The 4 Hr Rhythmmentioning

confidence: 99%

“…Nevertheless, the basic pacemaker of biological clocks appears to reside on a cellular level and is integrated into molecular networks of the cell. Studies in cultured cells have significantly advanced 15 our understanding of the molecular mechanisms of the cell cycle and, more recently, of other cellautonomous biological clocks. To detect periodicity in cell behavior, the expression of specific genes is usually measured by biochemical sampling or by using specific luminescent markers.…”

Section: Introductionmentioning

confidence: 99%

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A New Ultradian Rhythm Linked to Protein Degradation and Synthesis in Mammalian Cells

Ghenim

Allier

Obeïd

et al. 2019

Preprint

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We describe a new ultradian rhythm that occurs during the interphase of the cell cycle in a wide range of mammalian cells, including both primary and transformed cells. The rhythm 15 was detected by holographic lens-free microscopy that follows the individual histories of the dry mass of thousands of live cells simultaneously, each at a resolution of five minutes. Importantly, the rhythm was observed in inherently heterogeneous cell populations, thus eliminating synchronization and labeling bias. The rhythm is independent of circadian rhythm, has a period of 4 hours and is temperature-compensated. We demonstrated that the 4 hr rhythm is suppressed by 20 2 proteostasis disruptors and is detected only in proliferating cells, suggesting that it represents the periodic dynamics of protein mass in growing cells. Brief teaser:We have revealed a 4 hr rhythm in cell dry mass dynamics that seems to be general in proliferating mammalian cells.

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Section: Role Of Cell Cycle and Circadian Oscillatorsmentioning

confidence: 60%

Section: Role Of Cell Cycle and Circadian Oscillatorsmentioning

confidence: 67%

“…1A. We used lens-free microscopy to 15 analyze a large number of cells over several days by taking holographic images every 5 minutes.…”

Section: The 4 Hr Rhythmmentioning

confidence: 99%

Section: The 4 Hr Rhythmmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

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A New Ultradian Rhythm Linked to Protein Degradation and Synthesis in Mammalian Cells

Ghenim

Allier

Obeïd

et al. 2019

Preprint

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Protocols for in vitro reconstitution of the cyanobacterial circadian clock

et al. 2023

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Circadian clocks are intracellular systems that orchestrate metabolic processes in anticipation of sunrise and sunset by providing an internal representation of local time. Because the ~24‐h metabolic rhythms they produce are important to health across diverse life forms there is growing interest in their mechanisms. However, mechanistic studies are challenging in vivo due to the complex, that is, poorly defined, milieu of live cells. Recently, we reconstituted the intact circadian clock of cyanobacteria in vitro. It oscillates autonomously and remains phase coherent for many days with a fluorescence‐based readout that enables real‐time observation of individual clock proteins and promoter DNA simultaneously under defined conditions without user intervention. We found that reproducibility of the reactions required strict adherence to the quality of each recombinant clock protein purified from Escherichia coli. Here, we provide protocols for preparing in vitro clock samples so that other labs can ask questions about how changing environments, like temperature, metabolites, and protein levels are reflected in the core oscillator and propagated to regulation of transcription, providing deeper mechanistic insights into clock biology.

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