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Circadian Cryptochrome 1 (CRY1) Rhythmically Modulates the DNA Damage Response [ChIP-seq]

ABSTRACT: These studies identify a novel node of AR-regulated DNA repair through CRY1 directly regulating HR-mediated DNA repair to enhance cancer progression and discern a potential avenue to be targeted in late stage disease to improve patient outcome.

ORGANISM(S): Homo sapiens

PROVIDER: GSE144960 | GEO | 2020/11/24

REPOSITORIES: GEO

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Circadian Cryptochrome 1 (CRY1) Rhythmically Modulates the DNA Damage Response [RNA-seq]

Project description:These studies identify a novel node of AR-regulated DNA repair through CRY1 directly regulating HR-mediated DNA repair to enhance cancer progression and discern a potential avenue to be targeted in late stage disease to improve patient outcome.

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Circadian Cryptochrome 1 (CRY1) Rhythmically Modulates the DNA Damage Response

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Circadian Cryptochrome 1 (CRY1) Rhythmically Modulates the DNA Damage Response [ChIP-seq]

Project description:Circadian Cryptochrome 1 (CRY1) Rhythmically Modulates the DNA Damage Response [ChIP-seq]

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Circadian Cryptochrome 1 (CRY1) Rhythmically Modulates the DNA Damage Response

Project description:This SuperSeries is composed of the SubSeries listed below.

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ChIP-seq of three core components of the circadian clock (BMAL1, CLOCK and CRY1) in unsynchronized human U2OS cells

Project description:The human osteosarcoma cell line U2OS contains a functional circadian clock but expresses only a few rhythmic genes. We identified by ChIP-seq analysis 3040 binding sites of the circadian transcription factors BMAL1, CLOCK, and CRY1 in the genome of U2OS cells, comparable to the number found in highly rhythmic tissues like liver. ChIP was performed as described previously (Rey et al, 2011; doi:10.1371/journal.pbio.1000595) using confluent desynchronized U2OS cells and BMAL1, CLOCK, and CRY1-antibodies. Immunoprecipitated chromatin (10 ng DNA of 8 independent BMAL1 ChIPs with enrichment levels > 80-fold, 9 ng DNA of a CRY1 ChIP with 10-fold enrichment, and 8 ng DNA of a CLOCK ChIP with 40-fold enrichment) was used for library preparation. Three lanes of the BMAL1 library, and one lane each of the CRY1 and CLOCK library were sequenced on the Illumina Genome Analyzer IIx using Single-Read Cluster Generation Kit and 36 Cycle Sequencing Kit v2 (Lausanne Genomics Technologies Facility).

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The circadian clock protein Cryptochrome 1 is a direct target and feedback regulator of the Hippo pathway

Project description:To examine the impact of Cry1 downregulation on gene expression, empty vector (Sh Ctrl) and Sh-RNA targeting Cry1 (Sh Cry1) were used to produce lentiviral vector. Cell were then infected with control and sh-RNA lentivirus and then selected with puromycin. Upon validation of the knockdown with real time PCR. RNA were prepared from cells stably expressing empty vector or sh-RNA against Cry1 and used gene expression profiling by RNA seq. RNA of two biological replicate were prepared from each genotype.

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The circadian cryptochrome, CRY1, is a pro-tumorigenic factor that rhythmically modulates DNA repair.

Project description:Mechanisms regulating DNA repair processes remain incompletely defined. Here, the circadian factor CRY1, an evolutionally conserved transcriptional coregulator, is identified as a tumor specific regulator of DNA repair. Key findings demonstrate that CRY1 expression is androgen-responsive and associates with poor outcome in prostate cancer. Functional studies and first-in-field mapping of the CRY1 cistrome and transcriptome reveal that CRY1 regulates DNA repair and the G2/M transition. DNA damage stabilizes CRY1 in cancer (in vitro, in vivo, and human tumors ex vivo), which proves critical for efficient DNA repair. Further mechanistic investigation shows that stabilized CRY1 temporally regulates expression of genes required for homologous recombination. Collectively, these findings reveal that CRY1 is hormone-induced in tumors, is further stabilized by genomic insult, and promotes DNA repair and cell survival through temporal transcriptional regulation. These studies identify the circadian factor CRY1 as pro-tumorigenic and nominate CRY1 as a new therapeutic target.

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DNA damage shifts circadian clock time via Hausp-dependent Cry1 stabilization.

Project description:The circadian transcriptional repressors cryptochrome 1 (Cry1) and 2 (Cry2) evolved from photolyases, bacterial light-activated DNA repair enzymes. In this study, we report that while they have lost DNA repair activity, Cry1/2 adapted to protect genomic integrity by responding to DNA damage through posttranslational modification and coordinating the downstream transcriptional response. We demonstrate that genotoxic stress stimulates Cry1 phosphorylation and its deubiquitination by Herpes virus associated ubiquitin-specific protease (Hausp, a.k.a Usp7), stabilizing Cry1 and shifting circadian clock time. DNA damage also increases Cry2 interaction with Fbxl3, destabilizing Cry2. Thus, genotoxic stress increases the Cry1/Cry2 ratio, suggesting distinct functions for Cry1 and Cry2 following DNA damage. Indeed, the transcriptional response to genotoxic stress is enhanced in Cry1-/- and blunted in Cry2-/- cells. Furthermore, Cry2-/- cells accumulate damaged DNA. These results suggest that Cry1 and Cry2, which evolved from DNA repair enzymes, protect genomic integrity via coordinated transcriptional regulation.

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AP-MS of small molecule that selectively destabilizes Cryptochrome 1

Project description:Cryptochromes are negative transcriptional regulators of the circadian clock in mammals. It is not clear how reducing the level of endogenous CRY1 in mammals will affect circadian rhythm and the relation of such a decrease with apoptosis. Here, we discovered a molecule (M47) that destabilizes Cryptochrome 1 (CRY1) both in vitro and in vivo. The M47 selectively enhanced the degradation rate of CRY1 by increasing its ubiquitination and resulted in increasing the circadian period length of U2OS Bmal1-dLuc cells. In addition, subcellular fractionation studies from mice liver indicated that M47 increased degradation of the CRY1 in the nucleus. Furthermore, M47-mediated CRY1 reduction enhanced cisplatin-induced apoptosis in Ras-transformed p53 null fibroblast cells. Systemic repetitive administration of M47 increased the median lifespan of p53-/- mice by ~25%. Collectively our data suggest that M47 is a very promising molecule to treat forms of cancer depending on the p53 mutation.

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