Circadian clock, carcinogenesis, chronochemotherapy connections

Journal of Biological Chemistry, Год журнала: 2021, Номер 297(3), С. 101068 - 101068

Опубликована: Авг. 8, 2021

The circadian clock controls the expression of nearly 50% protein coding genes in mice and most likely humans as well. Therefore, disruption is presumed to have serious pathological effects including cancer. However, epidemiological studies on individuals with because night shift or rotating work produced contradictory data not conducive scientific consensus whether increases incidence breast, ovarian, prostate, colorectal cancers. Similarly, genetically engineered do exhibit spontaneous radiation-induced cancers at higher than wild-type controls. Because many cellular functions cell cycle division are, least part, controlled by molecular components (CLOCK, BMAL1, CRYs, PERs), it has also been expected that appropriate timing chemotherapy may increase efficacy chemotherapeutic drugs ameliorate their side effect. empirical attempts chronochemotherapy beneficial outcomes. Using without human tumor xenografts, sites DNA damage repair following treatment anticancer drug cisplatin mapped genome-wide single nucleotide resolution a function time. indicate mechanism-based such these provide information necessary for devising rational regimens.

Язык: Английский

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Loss of Timeless Underlies an Evolutionary Transition within the Circadian Clock

Molecular Biology and Evolution, Год журнала: 2021, Номер 39(1)

Опубликована: Дек. 2, 2021

Abstract Most organisms possess time-keeping devices called circadian clocks. At the molecular level, clocks consist of transcription–translation feedback loops (TTFLs). Although some components negative TTFL are conserved across animals, important differences exist between typical models, such as mouse and fruit fly. In Drosophila, key PERIOD (PER) TIMELESS (TIM-d) proteins, whereas mammalian clock relies on PER CRYPTOCHROME (CRY-m). Importantly, how has maintained functionality during evolutionary transitions different states remains elusive. Therefore, we systematically described gene setup in major bilaterian lineages identified marked lineage-specific their constitution. Then performed a thorough functional analysis linden bug Pyrrhocoris apterus, an insect species comprising features characteristic both Drosophila Unexpectedly, knockout timeless-d, essential for ticking did not compromise rhythmicity P. it only accelerated its pace. Furthermore, silencing timeless-m, ancestral timeless type ubiquitously present resulted mild gradual loss rhythmicity, supporting possible participation clock, which is consistent with timeless-m role suggested by research models. The dispensability timeless-d apterus allows drawing scenario remained at each step transition from state to TIM-d-independent + CRY-m system operating extant vertebrates, including humans.

Язык: Английский

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PERspectives on circadian cell biology

Philosophical Transactions of the Royal Society B Biological Sciences, Год журнала: 2025, Номер 380(1918)

Опубликована: Янв. 23, 2025

Daily rhythms in the activities of PERIOD proteins are critical to temporal regulation mammalian physiology. While molecular partners and genetic circuits that allow effect auto-repression regulate transcriptional programmes increasingly well understood, comprehension time-resolved mechanisms conduct this daily dance is incomplete. Here, we consider character controversies central clock protein with a focus on its intrinsically disordered nature. This article part Theo Murphy meeting issue ‘Circadian infection immunity’.

Язык: Английский

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Chronic CRYPTOCHROME deficiency enhances cell-intrinsic antiviral defences

Philosophical Transactions of the Royal Society B Biological Sciences, Год журнала: 2025, Номер 380(1918)

Опубликована: Янв. 23, 2025

The within-host environment changes over circadian time and influences the replication severity of viruses. Genetic knockout transcription factors CRYPTOCHROME 1 2 ( CRY1 −/− / CRY2 ; CKO) leads to altered protein homeostasis chronic activation integrated stress response (ISR). adaptive ISR signalling pathways help restore cellular by downregulating synthesis in endoplasmic reticulum overloading or viral infections. By quantitative mass spectrometry analysis, we reveal that many recognition proteins type I interferon (IFN) effectors are significantly upregulated lung fibroblast cells from CKO mice compared with wild-type (WT) mice. This basal ‘antiviral state’ restricts growth influenza A virus is governed interaction between proteotoxic constitutive IFN signalling. proteome composition signature were partially phenocopied upon sustained depletion (CRY) using a small-molecule CRY degrader, modest differential gene expression consistent differences seen WT cells. Our results highlight crosstalk rhythms, cell-intrinsic antiviral defences homeostasis, providing tractable molecular model investigate interface these key contributors human health disease. article part Theo Murphy meeting issue ‘Circadian rhythms infection immunity’.

Язык: Английский

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CRYPTOCHROMES confer robustness, not rhythmicity, to circadian timekeeping

The EMBO Journal, Год журнала: 2021, Номер 40(7)

Опубликована: Янв. 25, 2021

Article25 January 2021Open Access Source DataTransparent process CRYPTOCHROMES confer robustness, not rhythmicity, to circadian timekeeping Marrit Putker orcid.org/0000-0001-9290-408X MRC Laboratory of Molecular Biology, Cambridge, UK Search for more papers by this author David C S Wong orcid.org/0000-0002-1712-9527 Estere Seinkmane orcid.org/0000-0002-3636-4709 Nina M Rzechorzek orcid.org/0000-0003-3209-5019 Aiwei Zeng orcid.org/0000-0003-0354-2529 Nathaniel P Hoyle Johanna E Chesham orcid.org/0000-0002-8981-2667 Mathew D Edwards orcid.org/0000-0002-3573-0025 Kevin A Feeney orcid.org/0000-0003-3143-818X Robin Fischer Biozentrum Universität, Würzburg, Germany Nicolai Peschel orcid.org/0000-0003-3488-832X Ko-Fan Chen orcid.org/0000-0002-7305-6254 Institute Neurology, University College London, Michael Vanden Oever orcid.org/0000-0002-2725-6300 Faculty Medicine, Imperial Rachel Edgar orcid.org/0000-0002-3348-0851 Christopher Selby Department Biochemistry and Biophysics, North Carolina School Chapel Hill, NC, USA Aziz Sancar orcid.org/0000-0001-6469-4900 John O'Neill Corresponding Author [email protected] orcid.org/0000-0003-2204-6096 Information Putker1, Wong1, Seinkmane1, Rzechorzek1, Zeng1, Hoyle1, Chesham1, Edwards1,6, Feeney1, Fischer2, Peschel2, Chen3,7, Oever4, Edgar4, Selby5, Sancar5 *,1 1MRC 2Biozentrum 3Institute 4Faculty 5Department 6Present address: UCL Sainsbury Wellcome Centre Neural Circuits Behaviour, 7Present Genetics Genome Leicester, *Corresponding author. Tel: +44 7739 729425; E-mail: The EMBO Journal (2021)40:e106745https://doi.org/10.15252/embj.2020106745 PDFDownload PDF article text main figures. Peer ReviewDownload a summary the editorial decision including letters, reviewer comments responses feedback. ToolsAdd favoritesDownload CitationsTrack CitationsPermissions ShareFacebookTwitterLinked InMendeleyWechatReddit Figures & Info Abstract Circadian rhythms are pervasive property mammalian cells, tissues behaviour, ensuring physiological adaptation solar time. Models cellular revolve around transcriptional feedback repression, whereby CLOCK BMAL1 activate expression PERIOD (PER) CRYPTOCHROME (CRY), which in turn repress CLOCK/BMAL1 activity. CRY proteins therefore considered essential components clock mechanism, supported behavioural arrhythmicity CRY-deficient (CKO) mice under constant conditions. Challenging interpretation, we find locomotor adult CKO specific environmental conditions PER2 levels when is absent. CRY-less oscillations variable their have shorter periods than wild-type controls. Importantly, classic hallmarks such as temperature compensation period determination CK1δ/ε activity be maintained. In absence CRY-mediated repression rhythmic Per2 transcription, protein sustained several cycles, accompanied variation stability. We suggest that, whereas imparts robustness functionality onto biological clocks, core mechanism post-translational. SYNOPSIS (CRY) central regulators transcription/translation loop. finding that persists, albeit with reduced cells suggests gene determined evolutionarily-conserved post-translational timing mechanisms. dispensable but functions make robust. knockout exhibit rhythmicity abundance component amplified by, does require, transcription. CK1 GSK3 kinases regulate stability CRY. Introduction adaptive advantage conferred on organisms anticipation 24-h cycle day night has selected evolution clocks different molecular forms, present throughout all kingdoms life (Rosbash, 2009; et al, 2012). robust, they "capable performing without failure wide range conditions" (Merriam-Webster Dictionary, 2020). proposed generate daily delayed transcriptional–translational loop (TTFL) consists activating transcription factor complexes containing repressive complexes, BMAL1:CLOCK targets (reviewed Dunlap, 1999; Reppert Weaver, 2002; Takahashi, 2016). Various coupled, non-essential, auxiliary mechanisms thought fine-tune TTFL co-ordinate cell-type-specific temporal organisation programs; best characterised being effected E-box mediated REV-ERBα/β, encoded Nr1d1/2 genes (Preitner Ueda, 2007; Liu 2008; These loops sufficient 2008). CRY1 CRY2 operate semi-redundantly repressors (Ye 2014; Chiou 2016), required nuclear import PER proteins, together indispensable regulation vivo well cultured ex (Kume Sato 2006; 2016; Ode 2017). Certainly, homozygous null Cry1 Cry2 do express rest/activity cycles standard experimental (Thresher 1998; Horst Muijtjens, Vitaterna 1999). hypothalamic suprachiasmatic nucleus (SCN) locus co-ordination behaviour physiology, research over last two decades stressed strong correlation between SCN its (Welsh 2010; Anand 2013). were intrigued observation roughly half organotypic slices prepared from Cry1−/−,Cry2−/− (CRY knockout; CKO) mouse neonates continue ~ 20 h (short period) rhythms, observed using genetically PER2::LUC protein::luciferase fusion reporter (Maywood 2011; Ono 2013b), despite having previously been described arrhythmic (Liu 2007). Moreover, short reported raised birth light (Ono 2013a). As only neonatal vivo, suggested network-level SCN-specific rescue neuronal circuits, desynchronise during post-natal development 2013b). our view, however, these observations difficult reconcile an requirement generation rhythms. Rather, consistent making important contribution rhythm functional outputs, rather per se, recently shown Bmal1 Clock (Landgraf Ray 2020), had both individual (Bunger 2000; DeBruyne This further reports constitutive perturbs abolish (Fan Nangle Recent questioned need enable timekeeping. For example, translation regulated cytosolic through transcription-independent (Lipton 2015), isolated erythrocytes lacking any DNA (O'Neill Reddy, Cho 2014). some species eukaryotic alga prokaryotic cyanobacteria can occur entirely post-translationally (Sweeney Haxo, 1961; Nakajima 2005; Tomita 2011). Whether non-transcriptional other (nucleated) unknown hence relationship TTFL-mediated open question. Here, used mice, widely accepted Ukai-Tadenuma 2016) test whether function remained might begin dissect postulated oscillator or "cytoscillator" (Hastings Results Cell-autonomous Consistent previous observations, found no significant following entrainment 12 h:12 light:dark (LD) (LL). Upon transition darkness (DD) [described stronger zeitgeber (Chen 2008)] expressed bouts consolidated average 17 greater variance WT controls (Figs 1A B, EV1A–C). Fig 1, representative actograms plotted endogenous tau (τ) allow periodic rest–activity readily observed; 24-h-plotted Figs EV1A EV2A. showed significantly amplitude compared (WT) controls, persisted > 2 weeks, possessing residual oscillation entrained (Fig EV2-EV5 Appendix). support accordance longitudinal bioluminescence recordings revealed approximately 40% 1C). line data reports, exhibited 1D). Figure 1. CRY-independent occurs cell-autonomously Representative double-plotted showing wheel-running (yellow shading) thereafter darkness. Note 48 x-axis vs. 32 CKO. Full figure modulo 24 presented EV1A. Mean (± SEM) (n = 4). P-values calculated two-way ANOVA. Longitudinal (black) (red) (RLU; relative units). traces. immortalised lung fibroblasts. Left panel shows raw traces recording, right same detrended moving remove differences baseline expression. Period fibroblast at least 31 experiments ≥ 3 experiment, values ± SEM shown). unpaired t-test Welch correction. Standard deviations differ (F-test: < 0.0001). Download PowerPoint Click here expand figure. EV1. shading indicating lights on) (top) (LL) (bottom) (DD). Top four figures (modulo h). Rhythmic LL DD condition becomes clear plotting 16-h (i.e. LD also 1A. Bottom left, periodograms weeks darkness, either light. second cohort highlights (left, horizontal represents mean, F-test variance); (right) 10) 12) 1 week (mean SEM, 2-way ANOVA Sidak's MCT). Examples independent fibroblasts variability shape Two experiment. Stringent entrainment, e.g. dexamethasone, increases likelihood observing 30% did observe clearly cycles. Despite efforts, many years, unable identify set recording consistently produced forced conclude variables play adequately able control for. Genotyping study. Left: PCR genotyping expected pattern knockout. Right: Western blot analysis whole cell lysates (WCL) probed antibodies against CRY2. Interexperimental comparison Paired means 1F where rhythmic. paired t-test. Intraexperimental replicates) SEM). P-value was Damping rates example EV1E damped sine wave fitting n 106, 109). available online EV2. Entrainment deficiency Timeless-independent Drosophila melanogaster A. 5) cannot 8 h:8 entrain light-dark 5). Equal numbers age-matched male female used. C. dominant zeitgeber, vary contrast, unnatural indicates vivo. Two-way significance multiple comparisons reported; mean. D. Normalised, XLG-luciferase (XLG-LUC; equivalent PER2::LUC) timeless (Timout) flies (detrended SEM; 21, Timout 36). difference y-axis scaling. E. Damped fit (D). (extra sum-of-squares F-test) indicate hypothesis (straight line). F. Significant XLG-LUC flies. t-test, 36. Data information: Lamaze al (2017). Similar behaviourally cycles: https://opus.bibliothek.uni-wuerzburg.de/frontdoor/index/index/year/2015/docId/11914 EV3. post-transcriptionally curve recombinant luciferase determine number molecules. Known concentrations spiked into (non-luciferase containing) lysate reproduce conditions, signal measured 20-s integration time (CP s; counts s). fitted straight (red line, 95% CI). grey dotted lines (linear) area molecules experiment 3A. co-immunoprecipitation samples 3B. (IgG) pulldowns performed peak (determined parallel recordings) technical replicates (A–C). blots mRNA qPCR one 2, mean timeseries preferentially F-test, 0.0321), (ns). co-recording cultures depicted 3C. Detrended Cry1:LUC (MAFs) 4, extra (P 0.0001), could (no P-value). Nr1d1 promoter WT, quadruple Cry1/2-Per1/2 (CPKO) embryonic (MEFs) recorded 32°C, 3, (solid) ±SEM (dashed). (not significant, temperature-compensated 3). Expanded view Per2:LUC show detected C(P)KO MEFs, Zoomed-out version 37°C 3D 32°C EV3E. temperature-compensated, (from h) Nr1d1:LUC demonstrate explain EV4. oscillates assayed puromycin incorporation Cells synchronised harvested every after 10-min pulse (10 µg/ml). Incorporation blotting anti-puromycin antibody. quantified corrected total loading (Coomassie staining). Bioluminescence labelling course genotypes. immunoprecipitation S6K, eIF4a BMAL1. dexamethasone synchronisation, immunoprecipitated. Example (left) SV40::LUC pulsed 10 µM CHX 46 recording. resulting (symbols) one-phase decay (blue Multiple stable basal treated 25 μg/ml allowing turnover inferred signal. panel, decline simple exponential (solid lines) derive half-life each line. Right level reported, F-test). Timing pulses (labelled I-II (cycle 1) b 2)), trace corresponding G H. phases two-tailed Half-life line-fitting cells. EV5. role cytoplasmic presence inhibitor PF670462 (0.3 µM; PF), 5A (A), CHIR99021 (5 CHIR). KL001 (1 µM). respective quantifications CPKO Nr1d1::LUC (0.1 µM) (3 CHIR) (genotype drug interaction effect) reported. explanations account phenotype: (i) explanation: genetic loss compensated network circuits whose sensitive developmental phase small variations slice preparation Evans 2012; 2013b; Tokuda 2015); (ii) cell-intrinsic (or observed) stochastically less counterparts, interneuronal signalling To distinguish possibilities, asked populations fibroblasts, lack specialised neuropeptidergic so case 1E, EV1C D). Across 100 recordings, independently generated (male female), days Again, increased within (F-test 0.0001, 1F, F). results, occurred experiments, assays. there very little occurrence w

Язык: Английский

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Endogenous circadian reporters reveal functional differences of PERIOD paralogs and the significance of PERIOD:CK1 stable interaction

Proceedings of the National Academy of Sciences, Год журнала: 2023, Номер 120(6)

Опубликована: Фев. 1, 2023

Adverse consequences from having a faulty circadian clock include compromised sleep quality and poor performance in the short-term, metabolic diseases cancer long-term. However, our understanding of disorders is limited by incompleteness molecular models dearth defined mutant models. Because it would be prohibitively expensive to develop live animal study full range complicated mechanisms, we developed PER1-luc PER2-luc endogenous reporters validated cell model, U-2 OS, where genome can easily manipulated, functional mutations accurately studied. When major genes were knocked out these cells, rhythms modulated similarly compared with corresponding mice, validating platform for genetics studies. Using reporter uncovered critical differences between two paralogs PER . Although PER1 PER2 are considered redundant either one serve as pacemaker alone, they dramatically different biochemical parameters such stability phosphorylation kinetics. Consistently, phase was knockout cells. We further showed that stable binding casein kinase1δ/ε not required itself, but delayed timing phosphorylation. Our system used an efficient associated pathogenic their underlying mechanisms.

Язык: Английский

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Autonomous clocks that regulate organelle biogenesis, cytoskeletal organization, and intracellular dynamics

eLife, Год журнала: 2021, Номер 10

Опубликована: Сен. 29, 2021

How do cells perceive time? Do use temporal information to regulate the production/degradation of their enzymes, membranes, and organelles? Does controlling biological time influence cytoskeletal organization cellular architecture in ways that confer evolutionary physiological advantages? Potential answers these fundamental questions cell biology have historically revolved around discussion 'master' programs, such as principal cyclin-dependent kinase/cyclin division oscillator circadian clock. In this review, we provide an overview recent evidence supporting emerging concept 'autonomous clocks,' which under normal conditions can be

Язык: Английский

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Circadian disruption in lung fibroblasts enhances NF‐κB activity to exacerbate neutrophil recruitment

The FASEB Journal, Год журнала: 2023, Номер 37(2)

Опубликована: Янв. 9, 2023

Abstract Fibroblasts are stromal cells abundant throughout tissues, including the lungs. integral coordinators of immune cell recruitment through chemokine secretion. Circadian rhythms direct to lung, which in turn impacts response infection and survival. Although fibroblasts display robust circadian rhythms, contribution fibroblast molecular clock lung‐specific migration remains be established. Mice challenged intranasally with lipopolysaccharide (LPS) at dusk showed increased expression pro‐inflammatory cytokine IL‐1β CXCL5 was accompanied by neutrophil recruitment. Primary lung knockdown core gene Bmal1 immortalized −/− also displayed Cxcl5 under stimulation. Conditioned media obtained from IL‐1β‐stimulated fibroblasts‐induced greater compared +/+ controls. Phosphorylation NF‐κB subunit, p65, enhanced fibroblasts, pharmacological inhibition attenuated production observed these cells. Collectively, results demonstrate that represses activity control during an inflammatory response.

Язык: Английский

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Ultradian rhythms of AKT phosphorylation and gene expression emerge in the absence of the circadian clock components Per1 and Per2

PLoS Biology, Год журнала: 2021, Номер 19(12), С. e3001492 - e3001492

Опубликована: Дек. 30, 2021

Rhythmicity of biological processes can be elicited either in response to environmental cycles or driven by endogenous oscillators. In mammals, the circadian clock drives about 24-hour rhythms multitude metabolic and physiological anticipation daily oscillations. Also at intersection environment metabolism is protein kinase—AKT. It conveys extracellular signals, primarily feeding-related regulate various key cellular functions. Previous studies mice identified rhythmicity AKT activation (pAKT) with elevated levels fed state. However, it still unknown whether rhythmic through intrinsic mechanisms. Here, we inspected temporal changes pAKT both cultured cells animal models. cells, showed oscillations similar those observed livers wild-type under free-running conditions. Unexpectedly, Per1 , 2 −/− but not Bmal1 detected ultradian (about 16 hours) levels. Importantly, liver transcriptome also rhythms, corresponding consisting AKT-related genes regulators. Overall, our findings reveal gene expression phosphorylation that emerge absence genes.

Язык: Английский

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The role of cell-autonomous circadian oscillation of Cry transcription in circadian rhythm generation

Cell Reports, Год журнала: 2022, Номер 39(3), С. 110703 - 110703

Опубликована: Апрель 1, 2022

The current model of the mammalian circadian clock describes cell-autonomous and negative feedback-driven oscillation Cry Per transcription as core rhythm generator. However, actual contribution this to generation remains undefined. Here we perform targeted disruption cis elements indispensable for oscillation. Mice lacking overt show robust rhythms in locomotor activity. In addition, tissue-autonomous are absence Unexpectedly, although leads severe attenuation at level, Per2 accumulation remain robust. As a mechanism explain counterintuitive result, half-life shows independent may therefore partially functional even because degradation.

Язык: Английский

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