title: Early doors (Edo) mutant mouse reveals the importance of period 2 (PER2) PAS domain structure for circadian pacemaking.
abstract: The suprachiasmatic nucleus (SCN) defines 24 h of time via a transcriptional/posttranslational feedback loop in which transactivation of Per (period) and Cry (cryptochrome) genes by BMAL1-CLOCK complexes is suppressed by PER-CRY complexes. The molecular/structural basis of how circadian protein complexes function is poorly understood. We describe a novel N-ethyl-N-nitrosourea (ENU)-induced mutation, early doors (Edo), in the PER-ARNT-SIM (PAS) domain dimerization region of period 2 (PER2) (I324N) that accelerates the circadian clock of Per2(Edo/Edo) mice by 1.5 h. Structural and biophysical analyses revealed that Edo alters the packing of the highly conserved interdomain linker of the PER2 PAS core such that, although PER2(Edo) complexes with clock proteins, its vulnerability to degradation mediated by casein kinase 1ε (CSNK1E) is increased. The functional relevance of this mutation is revealed by the ultrashort (<19 h) but robust circadian rhythms in Per2(Edo/Edo); Csnk1e(Tau/Tau) mice and the SCN. These periods are unprecedented in mice. Thus, Per2(Edo) reveals a direct causal link between the molecular structure of the PER2 PAS core and the pace of SCN circadian timekeeping.
authors.affiliation: Medical Research Council Harwell, Harwell Science and Innovation Campus, Oxfordshire OX11 0RD, United Kingdom;
authors.forename: Stefania
authors.initials: S
authors.lastname: Militi
authors.affiliation: Division of Neurobiology, Medical Research Council Laboratory of Molecular Biology, Cambridge Biomedical Campus, Cambridge CB2 0QH, United Kingdom;
authors.forename: Elizabeth S
authors.initials: ES
authors.lastname: Maywood
authors.affiliation: Department of Chemistry and Biochemistry, University of California, Santa Cruz, CA 95064.
authors.forename: Colby R
authors.initials: CR
authors.lastname: Sandate
authors.affiliation: Division of Neurobiology, Medical Research Council Laboratory of Molecular Biology, Cambridge Biomedical Campus, Cambridge CB2 0QH, United Kingdom;
authors.forename: Johanna E
authors.initials: JE
authors.lastname: Chesham
authors.affiliation: Medical Research Council Harwell, Harwell Science and Innovation Campus, Oxfordshire OX11 0RD, United Kingdom;
authors.forename: Alun R
authors.identifier: http://orcid.org/0000-0002-0662-1073
authors.initials: AR
authors.lastname: Barnard
authors.affiliation: Medical Research Council Harwell, Harwell Science and Innovation Campus, Oxfordshire OX11 0RD, United Kingdom;
authors.forename: Michael J
authors.initials: MJ
authors.lastname: Parsons
authors.affiliation: Medical Research Council Harwell, Harwell Science and Innovation Campus, Oxfordshire OX11 0RD, United Kingdom;
authors.forename: Jennifer L
authors.initials: JL
authors.lastname: Vibert
authors.affiliation: Medical Research Council Harwell, Harwell Science and Innovation Campus, Oxfordshire OX11 0RD, United Kingdom;
authors.forename: Greg M
authors.initials: GM
authors.lastname: Joynson
authors.affiliation: Department of Chemistry and Biochemistry, University of California, Santa Cruz, CA 95064.
authors.forename: Carrie L
authors.initials: CL
authors.lastname: Partch
authors.affiliation: Division of Neurobiology, Medical Research Council Laboratory of Molecular Biology, Cambridge Biomedical Campus, Cambridge CB2 0QH, United Kingdom;
authors.forename: Michael H
authors.initials: MH
authors.lastname: Hastings
authors.affiliation: Medical Research Council Harwell, Harwell Science and Innovation Campus, Oxfordshire OX11 0RD, United Kingdom; p.nolan@har.mrc.ac.uk.
authors.forename: Patrick M
authors.initials: PM
authors.lastname: Nolan
mesh_terms: D000595:Amino Acid Sequence; D000818:Animals; D015153:Blotting, Western; D019556:COS Cells; D048149:Casein Kinase 1 epsilon / Q000235:genetics / Q000378:metabolism; D002522:Chlorocebus aethiops; D057906:Circadian Clocks / Q000235:genetics* / Q000502:physiology; D002940:Circadian Rhythm / Q000235:genetics* / Q000502:physiology; D005260:Female; D057809:HEK293 Cells; D006801:Humans; D008297:Male; D008807:Mice, Inbred BALB C; D008809:Mice, Inbred C3H; D008810:Mice, Inbred C57BL; D018345:Mice, Knockout; D008958:Models, Molecular; D008969:Molecular Sequence Data; D009043:Motor Activity / Q000235:genetics / Q000502:physiology; D020125:Mutation, Missense; D056950:Period Circadian Proteins / Q000737:chemistry / Q000235:genetics* / Q000378:metabolism; D055503:Protein Multimerization; D017434:Protein Structure, Tertiary; D017386:Sequence Homology, Amino Acid; D013493:Suprachiasmatic Nucleus / Q000378:metabolism / Q000503:physiopathology
chemical_list: C513312:Per2 protein, mouse; D056950:Period Circadian Proteins; D048149:Casein Kinase 1 epsilon
keywords: behavior; circadian period; genetic interaction; mouse mutant; protein stability