The Arg-293 of Cryptochrome1 is responsible for the allosteric regulation of CLOCK-CRY1 binding in circadian rhythm


Gul Ş., AYDIN C., Ozcan O., Gurkan B., Surme S., Baris I., ...More

JOURNAL OF BIOLOGICAL CHEMISTRY, vol.295, no.50, pp.17187-17199, 2020 (Peer-Reviewed Journal) identifier identifier identifier

  • Publication Type: Article / Article
  • Volume: 295 Issue: 50
  • Publication Date: 2020
  • Doi Number: 10.1074/jbc.ra120.014333
  • Journal Name: JOURNAL OF BIOLOGICAL CHEMISTRY
  • Journal Indexes: Science Citation Index Expanded, Scopus, Academic Search Premier, Applied Science & Technology Source, Aquatic Science & Fisheries Abstracts (ASFA), Artic & Antarctic Regions, BIOSIS, CAB Abstracts, Compendex, EMBASE, MEDLINE, Veterinary Science Database
  • Page Numbers: pp.17187-17199
  • Keywords: circadian rhythm, gene regulation, Cryptochrome 1, allostery, CLOCK, clock gene, cryptochrome, allosteric regulation, FEEDBACK REPRESSION, GENETIC-VARIANTS, MAMMALIAN CRY1, DNA-REPAIR, PROTEIN, IDENTIFICATION, ARCHITECTURE, DEGRADATION, ASSOCIATION, MUTATION

Abstract

Mammalian circadian clocks are driven by transcription/translation feedback loops composed of positive transcriptional activators (BMAL1 and CLOCK) and negative repressors (CRYPTOCHROMEs (CRYs) and PERIODs (PERs)). CRYs, in complex with PERs, bind to the BMAL1/CLOCK complex and repress E-box-driven transcription of clock-associated genes. There are two individual CRYs, with CRY1 exhibiting higher affinity to the BMAL1/CLOCK complex than CRY2. It is known that this differential binding is regulated by a dynamic serine-rich loop adjacent to the secondary pocket of both CRYs, but the underlying features controlling loop dynamics are not known. Here we report that allosteric regulation of the serine-rich loop is mediated by Arg-293 of CRY1, identified as a rare CRY1 SNP in the Ensembl and 1000 Genomes databases. The p.Arg293His CRY1 variant caused a shortened circadian period in a Cry1(-/-)Cry2(-/-) double knockout mouse embryonic fibroblast cell line. Moreover, the variant displayed reduced repressor activity on BMAL1/CLOCK driven transcription, which is explained by reduced affinity to BMAL1/CLOCK in the absence of PER2 compared with CRY1. Molecular dynamics simulations revealed that the p.Arg293His CRY1 variant altered a communication pathway between Arg-293 and the serine loop by reducing its dynamicity. Collectively, this study provides direct evidence that allosterism in CRY1 is critical for the regulation of circadian rhythm.