Limits...
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.

Show MeSH

Related in: MedlinePlus

Conserved SQ/TQ motifs present in Cry1 and/or Cry2.Sequence alignment of mouse and human Cry1 and Cry2 indicating the positions and conservation of several SQ/TQ motifs. (Numbers correspond to the amino acid positions in mouse Cry1.)DOI:http://dx.doi.org/10.7554/eLife.04883.016
© Copyright Policy
Related In: Results  -  Collection

License
getmorefigures.php?uid=PMC4352707&req=5

fig4s2: Conserved SQ/TQ motifs present in Cry1 and/or Cry2.Sequence alignment of mouse and human Cry1 and Cry2 indicating the positions and conservation of several SQ/TQ motifs. (Numbers correspond to the amino acid positions in mouse Cry1.)DOI:http://dx.doi.org/10.7554/eLife.04883.016

Mentions: ATM- and PPM1G-dependent dephosphorylation of serine 18 in the N-terminus of Hausp has been reported to drive the DNA damage dependent disruption of Hausp interaction with Mdm2 and MdmX (Khoronenkova et al., 2012). Conversely, S18 de-phosphorylation may increase Hausp–Cry1 association because mutation of S18 to the non-phosphorylatable amino acid alanine (S18A) increases interaction and mutation to aspartic acid, which is chemically similar to phospho-serine, decreases the interaction (Figure 4A). However, S18 dephosphorylation cannot fully explain DNA damage induction of Cry1–Hausp interaction as evidenced by persistent stimulated association between Cry1 and Hausp S18A after DNA damage. Intriguingly, we (Figure 4—figure supplement 1, Supplementary file 2) and others (Gao et al., 2013) find that Cry1 and Cry2 interact with kinases that are activated by DNA damage and phosphorylate serine or threonine followed by glutamine, (S/T)-Q (Kim et al., 1999; O'Neill et al., 2000). Cry1 and Cry2 contain several such sequences (Figure 4—figure supplement 2), including three serines in the Cry1 C-terminal tail that are not conserved in Cry2.10.7554/eLife.04883.014Figure 4.DNA damage induced signaling modulates interactions of Cry1/2, Hausp, and Fbxl3.


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)

Conserved SQ/TQ motifs present in Cry1 and/or Cry2.Sequence alignment of mouse and human Cry1 and Cry2 indicating the positions and conservation of several SQ/TQ motifs. (Numbers correspond to the amino acid positions in mouse Cry1.)DOI:http://dx.doi.org/10.7554/eLife.04883.016
© Copyright Policy
Related In: Results  -  Collection

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

fig4s2: Conserved SQ/TQ motifs present in Cry1 and/or Cry2.Sequence alignment of mouse and human Cry1 and Cry2 indicating the positions and conservation of several SQ/TQ motifs. (Numbers correspond to the amino acid positions in mouse Cry1.)DOI:http://dx.doi.org/10.7554/eLife.04883.016
Mentions: ATM- and PPM1G-dependent dephosphorylation of serine 18 in the N-terminus of Hausp has been reported to drive the DNA damage dependent disruption of Hausp interaction with Mdm2 and MdmX (Khoronenkova et al., 2012). Conversely, S18 de-phosphorylation may increase Hausp–Cry1 association because mutation of S18 to the non-phosphorylatable amino acid alanine (S18A) increases interaction and mutation to aspartic acid, which is chemically similar to phospho-serine, decreases the interaction (Figure 4A). However, S18 dephosphorylation cannot fully explain DNA damage induction of Cry1–Hausp interaction as evidenced by persistent stimulated association between Cry1 and Hausp S18A after DNA damage. Intriguingly, we (Figure 4—figure supplement 1, Supplementary file 2) and others (Gao et al., 2013) find that Cry1 and Cry2 interact with kinases that are activated by DNA damage and phosphorylate serine or threonine followed by glutamine, (S/T)-Q (Kim et al., 1999; O'Neill et al., 2000). Cry1 and Cry2 contain several such sequences (Figure 4—figure supplement 2), including three serines in the Cry1 C-terminal tail that are not conserved in Cry2.10.7554/eLife.04883.014Figure 4.DNA damage induced signaling modulates interactions of Cry1/2, Hausp, and Fbxl3.

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