Identification of two amino acids in the C-terminal domain of mouse CRY2 essential for PER2 interaction

• Published in: BMC molecular biology Vol. 11; no. 1; p. 69
• Main Authors: Ozber, Natali, Baris, Ibrahim, Tatlici, Gulnaz, Gur, Ibrahim, Kilinc, Seda, Unal, Evrim B, Kavakli, Ibrahim H
• Format: Journal Article
• Language: English
• Published: England BioMed Central Ltd 14-09-2010; BioMed Central
• Subjects: Amino Acid Sequence; Amino acids; Animals; ARNTL Transcription Factors - genetics; ARNTL Transcription Factors - metabolism; Binding sites; Casein; Cell Line; Circadian rhythm; Circadian Rhythm - genetics; Circadian rhythms; CLOCK Proteins - genetics; CLOCK Proteins - metabolism; Cloning; Cryptochromes - genetics; Cryptochromes - metabolism; DNA repair; Genetic aspects; Health aspects; Humans; Insects; Kinases; Light; Mammals; Mice; Molecular Sequence Data; Mutagenesis; Period Circadian Proteins - genetics; Period Circadian Proteins - metabolism; Physiological aspects; Proteins; Sequence Alignment; Two-Hybrid System Techniques
• ISSN: 1471-2199; 1471-2199
• DOI: 10.1186/1471-2199-11-69

Cryptochromes (CRYs) are a class of flavoprotein blue-light signaling receptors found in plants and animals, and they control plant development and the entrainment of circadian rhythms. They also act as integral parts of the central circadian oscillator in humans and other animals. In mammals, the CLOCK-BMAL1 heterodimer activates transcription of the Per and Cry genes as well as clock-regulated genes. The PER2 proteins interact with CRY and CKIε, and the resulting ternary complexes translocate into the nucleus, where they negatively regulate the transcription of Per and Cry core clock genes and other clock-regulated output genes. Recent studies have indicated that the extended C-termini of the mammalian CRYs, as compared to photolyase proteins, interact with PER proteins.
We identified a region on mCRY2 (between residues 493 and 512) responsible for direct physical interaction with mPER2 by mammalian two-hybrid and co-immunoprecipitation assays. Moreover, using oligonucleotide-based degenerate PCR, we discovered that mutation of Arg-501 and Lys-503 of mCRY2 within this C-terminal region totally abolishes interaction with PER2.
Our results identify mCRY2 amino acid residues that interact with the mPER2 binding region and suggest the potential for rational drug design to inhibit CRYs for specific therapeutic approaches.