title: Conserved amino acid residues in C-terminus of PERIOD 2 are involved in interaction with CRYPTOCHROME 1.
abstract: We investigated the amino acid sequences of rat PERIOD2 (rPER2) that are required for interaction with CRYPTOCHROME1 (CRY1) to understand the molecular mechanism of the circadian clock. Co-immunoprecipitation assays using various C-terminal fragments of rPER2 with internal deletions revealed that amino acid residues 1179-1198 are necessary for interaction with CRY1. To identify precisely which amino acid residues are responsible for the interaction, we substituted alanine for residues conserved among PER isoforms and species. We found that more than three mutations of conserved PER2 residues impaired not only binding to CRY1 but also subsequent nuclear translocation, although mutations of non-conserved residues did not affect interaction with CRY1. Thus, the conserved amino acid residues of 1179-1198 in PER2 are apparently responsible for binding to CRY1.
authors.affiliation: Institute for Biological Resources and Functions, National Institute of Advanced Industrial Science and Technology, Tsukuba Central 6, 1-1-1 Higashi, Tsukuba, Ibaraki 305-8566, Japan. tatsunosuke-tomita@aist.go.jp
authors.forename: Tatsunosuke
authors.initials: T
authors.lastname: Tomita
authors.forename: Koyomi
authors.initials: K
authors.lastname: Miyazaki
authors.forename: Yoshiaki
authors.initials: Y
authors.lastname: Onishi
authors.forename: Shinya
authors.initials: S
authors.lastname: Honda
authors.forename: Norio
authors.initials: N
authors.lastname: Ishida
authors.forename: Katsutaka
authors.initials: K
authors.lastname: Oishi
mesh_terms: D000595:Amino Acid Sequence; D000818:Animals; D019556:COS Cells; D002467:Cell Nucleus / Q000378:metabolism; D002522:Chlorocebus aethiops; D017124:Conserved Sequence; D056931:Cryptochromes / Q000235:genetics / Q000378:metabolism*; D007124:Immunoenzyme Techniques; D047468:Immunoprecipitation; D008156:Luciferases / Q000378:metabolism; D051379:Mice; D008969:Molecular Sequence Data; D009154:Mutation / Q000235:genetics*; D056950:Period Circadian Proteins / Q000235:genetics / Q000378:metabolism*; D017434:Protein Structure, Tertiary; D021381:Protein Transport; D051381:Rats; D017386:Sequence Homology, Amino Acid; D014667:Vasopressins / Q000235:genetics
chemical_list: C538965:Cry1 protein, rat; D056931:Cryptochromes; C513311:Per2 protein, rat; D056950:Period Circadian Proteins; D014667:Vasopressins; D008156:Luciferases