An Isoform-Selective Modulator of Cryptochrome 1 Regulates Circadian Rhythms in Mammals

Simon Miller; Yoshiki Aikawa; Akiko Sugiyama; Yoshiko Nagai; Aya Hara; Tsuyoshi Oshima; Kazuma Amaike; Steve A Kay; Kenichiro Itami; Tsuyoshi Hirota

doi:10.1016/j.chembiol.2020.05.008

An Isoform-Selective Modulator of Cryptochrome 1 Regulates Circadian Rhythms in Mammals

Cell Chem Biol. 2020 Sep 17;27(9):1192-1198.e5. doi: 10.1016/j.chembiol.2020.05.008. Epub 2020 Jun 4.

Authors

Affiliations

¹ Institute of Transformative Bio-Molecules, Nagoya University, Nagoya 464-8601, Japan.
² Institute of Transformative Bio-Molecules, Nagoya University, Nagoya 464-8601, Japan; Department of Chemistry, Graduate School of Science, Nagoya University, Nagoya 464-8601, Japan.
³ Department of Neurology, Keck School of Medicine, University of Southern California, Los Angeles, CA 90089, USA.
⁴ Institute of Transformative Bio-Molecules, Nagoya University, Nagoya 464-8601, Japan. Electronic address: thirota@itbm.nagoya-u.ac.jp.

PMID: 32502390
DOI: 10.1016/j.chembiol.2020.05.008

Abstract

Cryptochrome 1 (CRY1) and CRY2 are core regulators of the circadian clock, and the development of isoform-selective modulators is important for the elucidation of their redundant and distinct functions. Here, we report the identification and functional characterization of a small-molecule modulator of the mammalian circadian clock that selectively controls CRY1. Cell-based circadian chemical screening identified a thienopyrimidine derivative KL201 that lengthened the period of circadian rhythms in cells and tissues. Functional assays revealed stabilization of CRY1 but not CRY2 by KL201. A structure-activity relationship study of KL201 derivatives in combination with X-ray crystallography of the CRY1-KL201 complex uncovered critical sites and interactions required for CRY1 regulation. KL201 bound to CRY1 in overlap with FBXL3, a subunit of ubiquitin ligase complex, and the effect of KL201 was blunted by knockdown of FBXL3. KL201 will facilitate isoform-selective regulation of CRY1 to accelerate chronobiology research and therapeutics against clock-related diseases.

Keywords: X-ray crystallography; chemical biology; circadian clock; cryptochrome; small-molecule modulator.

Publication types

Research Support, Non-U.S. Gov't

MeSH terms

ARNTL Transcription Factors / genetics
ARNTL Transcription Factors / metabolism
Binding Sites
Carbazoles / chemistry
Carbazoles / metabolism*
Carbazoles / pharmacology
Cell Line, Tumor
Circadian Rhythm* / drug effects
Cryptochromes / chemistry
Cryptochromes / genetics
Cryptochromes / metabolism*
Crystallography, X-Ray
F-Box Proteins / metabolism
Genes, Reporter
Humans
Molecular Docking Simulation
Period Circadian Proteins / genetics
Period Circadian Proteins / metabolism
Protein Binding
Structure-Activity Relationship
Ubiquitination

Substances

ARNTL Transcription Factors
BMAL1 protein, human
CRY1 protein, human
CRY2 protein, human
Carbazoles
Cryptochromes
F-Box Proteins
FBXL3 protein, human
PER2 protein, human
Period Circadian Proteins