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

Papp SJ, Huber AL, Jordan SD, Kriebs A, Nguyen M, Moresco JJ, Yates JR, Lamia KA - Elife (2015)

Bottom Line: 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.Indeed, the transcriptional response to genotoxic stress is enhanced in Cry1-/- and blunted in Cry2-/- cells.Furthermore, Cry2-/- cells accumulate damaged DNA.

View Article: PubMed Central - PubMed

Affiliation: Department of Chemical Physiology, Scripps Research Institute, La Jolla, United States.

ABSTRACT
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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Effect of DNA damage on Cry1-Hausp interaction in transfected 293T cells.HAUSP-V5, FLAG-Cry1, and Actin were detected by IB in IPs or whole cell lysates (WCL) from 293T cells transfected with the indicated plasmids (by calcium phosphate method) and treated with doxorubicin.DOI:http://dx.doi.org/10.7554/eLife.04883.011
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fig3s1: Effect of DNA damage on Cry1-Hausp interaction in transfected 293T cells.HAUSP-V5, FLAG-Cry1, and Actin were detected by IB in IPs or whole cell lysates (WCL) from 293T cells transfected with the indicated plasmids (by calcium phosphate method) and treated with doxorubicin.DOI:http://dx.doi.org/10.7554/eLife.04883.011


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

Papp SJ, Huber AL, Jordan SD, Kriebs A, Nguyen M, Moresco JJ, Yates JR, Lamia KA - Elife (2015)

Effect of DNA damage on Cry1-Hausp interaction in transfected 293T cells.HAUSP-V5, FLAG-Cry1, and Actin were detected by IB in IPs or whole cell lysates (WCL) from 293T cells transfected with the indicated plasmids (by calcium phosphate method) and treated with doxorubicin.DOI:http://dx.doi.org/10.7554/eLife.04883.011
© Copyright Policy
Related In: Results  -  Collection

License
Show All Figures
getmorefigures.php?uid=PMC4352707&req=5

fig3s1: Effect of DNA damage on Cry1-Hausp interaction in transfected 293T cells.HAUSP-V5, FLAG-Cry1, and Actin were detected by IB in IPs or whole cell lysates (WCL) from 293T cells transfected with the indicated plasmids (by calcium phosphate method) and treated with doxorubicin.DOI:http://dx.doi.org/10.7554/eLife.04883.011
Bottom Line: 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.Indeed, the transcriptional response to genotoxic stress is enhanced in Cry1-/- and blunted in Cry2-/- cells.Furthermore, Cry2-/- cells accumulate damaged DNA.

View Article: PubMed Central - PubMed

Affiliation: Department of Chemical Physiology, Scripps Research Institute, La Jolla, United States.

ABSTRACT
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.

Show MeSH
Related in: MedlinePlus