title: The 5' upstream region of mPer1 gene contains two promoters and is responsible for circadian oscillation.
abstract: The mPer1 gene is assumed to be a key molecule in the regulation and functioning of the mammalian circadian clock, which is based on the oscillation generated by a transcription-(post)translation feedback loop of a set of clock genes [1]. Robust circadian oscillation and acute light-elicited induction of mPer1 mRNA expression have been observed in the suprachiasmatic nucleus (SCN), the mammalian circadian center [2] [3]. To investigate the mechanism underlying the complex regulation of mPer1 expression, we isolated and characterized the 5' upstream region of the mPer1 gene. Unexpectedly, we identified two promoters, each followed by alternative first exons of mPer1. Consistent with the presence of multiple E-boxes in the promoters, exon-specific in situ hybridization of the SCN established that both promoters function in circadian oscillation and in light-induction of mPer1 expression. Transgenic mice carrying the 5' upstream region of the mPer1 gene fused to the luciferase gene demonstrated that a DNA fragment carrying both promoter regions is sufficient to elicit striking circadian oscillation in the SCN and responsiveness to light. Moreover, luminescence in the SCN accurately mirrored the mPer1 transcriptional activity. These transgenic mice will be very useful for monitoring clock-specific mPer1 expression in intact organisms and to follow the circadian clock in real time.
authors.affiliation: Department of Anatomy and Brain Science, Kobe University School of Medicine, Kobe, Japan.
authors.forename: S
authors.initials: S
authors.lastname: Yamaguchi
authors.forename: S
authors.initials: S
authors.lastname: Mitsui
authors.forename: S
authors.initials: S
authors.lastname: Miyake
authors.forename: L
authors.initials: L
authors.lastname: Yan
authors.forename: H
authors.initials: H
authors.lastname: Onishi
authors.forename: K
authors.initials: K
authors.lastname: Yagita
authors.forename: M
authors.initials: M
authors.lastname: Suzuki
authors.forename: S
authors.initials: S
authors.lastname: Shibata
authors.forename: M
authors.initials: M
authors.lastname: Kobayashi
authors.forename: H
authors.initials: H
authors.lastname: Okamura
mesh_terms: D000818:Animals; D001483:Base Sequence; D018797:Cell Cycle Proteins; D002940:Circadian Rhythm / Q000235:genetics*; D017931:DNA Primers / Q000235:genetics; D005091:Exons; D015870:Gene Expression; D017930:Genes, Reporter; D017403:In Situ Hybridization; D008156:Luciferases / Q000235:genetics; D051379:Mice; D008822:Mice, Transgenic; D009687:Nuclear Proteins / Q000235:genetics*; D056950:Period Circadian Proteins; D011401:Promoter Regions, Genetic; D012333:RNA, Messenger / Q000235:genetics / Q000378:metabolism; D013493:Suprachiasmatic Nucleus / Q000378:metabolism
chemical_list: D018797:Cell Cycle Proteins; D017931:DNA Primers; D009687:Nuclear Proteins; C539023:Per1 protein, mouse; D056950:Period Circadian Proteins; D012333:RNA, Messenger; D008156:Luciferases