Identification of a Novel Cryptochrome Differentiating Domain Required for Feedback Repression in Circadian Clock Function

Khan, Sanjoy K.; Xu, Hongwei; Ukai-Tadenuma, Maki; Burton, Brittany N.; Wang, Yongmei; Ueda, Hiroki R.; Liu, Andrew C.

doi:10.1074/jbc.m112.368001

Cited by 70 publications

(111 citation statements)

References 57 publications

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“…They are also consistent with previous findings in mice showing that the C-terminal domain of CRY1, which interacts with the BMAL1 C terminus, regulates clock function but is not necessary for transcriptional repression (38,39).…”

Section: Discussionsupporting

confidence: 81%

Vertebrate-like CRYPTOCHROME 2 from monarch regulates circadian transcription via independent repression of CLOCK and BMAL1 activity

Zhang

Markert

Groves

et al. 2017

Proc. Natl. Acad. Sci. U.S.A.

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Circadian repression of CLOCK-BMAL1 by PERIOD and CRYPTOCHROME (CRY) in mammals lies at the core of the circadian timekeeping mechanism. CRY repression of CLOCK-BMAL1 and regulation of circadian period are proposed to rely primarily on competition for binding with coactivators on an α-helix located within the transactivation domain (TAD) of the BMAL1 C terminus. This model has, however, not been tested in vivo. Here, we applied CRISPR/Cas9-mediated mutagenesis in the monarch butterfly (Danaus plexippus), which possesses a vertebrate-like CRY (dpCRY2) and an ortholog of BMAL1, to show that insect CRY2 regulates circadian repression through TAD α-helix-dependent and -independent mechanisms. Monarch mutants lacking the BMAL1 C terminus including the TAD exhibited arrhythmic eclosion behavior. In contrast, mutants lacking the TAD α-helix but retaining the most distal C-terminal residues exhibited robust rhythms during the first day of constant darkness (DD1), albeit with a delayed peak of eclosion. Phase delay in this mutant on DD1 was exacerbated in the presence of a single functional allele of dpCry2, and rhythmicity was abolished in the absence of dpCRY2. Reporter assays in Drosophila S2 cells further revealed that dpCRY2 represses through two distinct mechanisms: a TADdependent mechanism that involves the dpBMAL1 TAD α-helix and dpCLK W328 and a TAD-independent mechanism involving dpCLK E333. Together, our results provide evidence for independent mechanisms of vertebrate-like CRY circadian regulation on the BMAL1 C terminus and the CLK PAS-B domain and demonstrate the importance of a BMAL1 TAD-independent mechanism for generating circadian rhythms in vivo.circadian clock | CRYPTOCHROME 2 | BMAL1 C terminus | CLOCK | CRISPR

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Section: Discussionsupporting

confidence: 81%

Vertebrate-like CRYPTOCHROME 2 from monarch regulates circadian transcription via independent repression of CLOCK and BMAL1 activity

Zhang

Markert

Groves

et al. 2017

Proc. Natl. Acad. Sci. U.S.A.

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“…This flexibility could play a role in regulating CLOCK:BMAL1 function, as one of these loops, located within the BMAL1 PAS-A domain, is modified by sumoylation to control CLOCK:BMAL1 activity (29). These data also lay the foundation for future studies on the role of the disordered CRY1 C-terminal extension, which controls circadian period and amplitude (42). Understanding the structural basis for mutual exclusivity or synergy of clock protein interactions will provide a framework to elucidate the mechanistic underpinnings of the transcriptionbased mammalian circadian clock.…”

Section: Discussionmentioning

confidence: 86%

Formation of a repressive complex in the mammalian circadian clock is mediated by the secondary pocket of CRY1

Michael

Fribourgh

Chelliah

et al. 2017

Proc. Natl. Acad. Sci. U.S.A.

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The basic helix–loop–helix PAS domain (bHLH-PAS) transcription factor CLOCK:BMAL1 (brain and muscle Arnt-like protein 1) sits at the core of the mammalian circadian transcription/translation feedback loop. Precise control of CLOCK:BMAL1 activity by coactivators and repressors establishes the ∼24-h periodicity of gene expression. Formation of a repressive complex, defined by the core clock proteins cryptochrome 1 (CRY1):CLOCK:BMAL1, plays an important role controlling the switch from repression to activation each day. Here we show that CRY1 binds directly to the PAS domain core of CLOCK:BMAL1, driven primarily by interaction with the CLOCK PAS-B domain. Integrative modeling and solution X-ray scattering studies unambiguously position a key loop of the CLOCK PAS-B domain in the secondary pocket of CRY1, analogous to the antenna chromophore-binding pocket of photolyase. CRY1 docks onto the transcription factor alongside the PAS domains, extending above the DNA-binding bHLH domain. Single point mutations at the interface on either CRY1 or CLOCK disrupt formation of the ternary complex, highlighting the importance of this interface for direct regulation of CLOCK:BMAL1 activity by CRY1.

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“…VOLUME 288 • NUMBER 49 • DECEMBER 6, 2013 portion of the CRY1 protein has been unclear, with some evidence that it is involved in nuclear localization or BMAL binding (7,8,68) and other evidence that it is dispensable for core clock function (6). However, even in the latter case, some role for the C terminus in period control was noted (6).…”

Section: Journal Of Biological Chemistrymentioning

confidence: 99%

Phosphorylation of the Cryptochrome 1 C-terminal Tail Regulates Circadian Period Length

Gao

Yoo

Lee

et al. 2013

Journal of Biological Chemistry

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Background: Cryptochromes (CRYs) are transcriptional repressors that are critical components of the circadian clock. Results: We have identified a phosphorylation site in the CRY1 tail that is negatively regulated by the DNA repair enzyme DNA-dependent protein kinase. Conclusion: Phosphorylation of CRY1 on Ser-588 increases its half-life and lengthens the circadian period. Significance: The C-terminal tail of CRY1 modulates period length.

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Identification of a Novel Cryptochrome Differentiating Domain Required for Feedback Repression in Circadian Clock Function

Cited by 70 publications

References 57 publications

Vertebrate-like CRYPTOCHROME 2 from monarch regulates circadian transcription via independent repression of CLOCK and BMAL1 activity

Vertebrate-like CRYPTOCHROME 2 from monarch regulates circadian transcription via independent repression of CLOCK and BMAL1 activity

Formation of a repressive complex in the mammalian circadian clock is mediated by the secondary pocket of CRY1

Phosphorylation of the Cryptochrome 1 C-terminal Tail Regulates Circadian Period Length

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