TY - JOUR
T1 - Natural antisense transcript of Period2, Per2AS, regulates the amplitude of the mouse circadian clock
AU - Mosig, Rebecca A.
AU - Castaneda, Allison N.
AU - Deslauriers, Jacob C.
AU - Frazier, Landon P.
AU - He, Kevin L.
AU - Maghzian, Naseem
AU - Pokharel, Aarati
AU - Schrier, Camille T.
AU - Zhu, Lily
AU - Koike, Nobuya
AU - Tyson, John J.
AU - Green, Carla B.
AU - Takahashi, Joseph S.
AU - Kojima, Shihoko
N1 - Funding Information:
We thank Dr. Ueli Schibler (Universit? de Gen?ve) for providing the Bmal1-luc cell line, Dr. Seung-Hee Yoo (University of Texas Health Science Center at Houston) for PER2::LUC MEFs, and Dr. Andrew Liu (University of Florida) for Per2 shRNA vectors (Ramanathan et al. 2014). We also thank Melissa Makris (Flow Cytometry Resource Laboratory, Department of Biomedical Sciences and Pathobiology, Center for Molecular Medicine and Infectious Diseases, Virginia-Maryland Regional College of Veterinary Medicine, Virginia Polytechnic Institute and State University), Dr. Nicolaas Baudoin and Dr. Daniela Cimini (Department of Biological Sciences, Virginia Polytechnic Institute and State University), and Tsubasa Toda and Akari Ueta (Department of Environmental and Life Sciences, Toyohashi University of Technology) for technical assistance. We also thank all the past an current members of the Green, Takahashi, and Kojima laboratories for invaluable discussion. This work was supported by the Luther and Alice Hamlett Undergraduate Research Support (to C.T.S), Howard Hughes Medical Institute (to J.S.T), and National Institutes of Health GM127122 (to C.B.G), and GM126223 (to S.K.). J.S.T. is an Investigator in the Howard Hughes Medical Institute.
Funding Information:
We thank Dr. Ueli Schibler (Université de Genève) for providing the Bmal1-luc cell line, Dr. Seung-Hee Yoo (University of Texas Health Science Center at Houston) for PER2::LUC MEFs, and Dr. Andrew Liu (University of Florida) for Per2 shRNA vectors (Ram-anathan et al. 2014). We also thank Melissa Makris (Flow Cytometry Resource Laboratory, Department of Biomedical Sciences and Pathobiology, Center for Molecular Medicine and Infectious Diseases, Virginia-Maryland Regional College of Veterinary Medicine, Virginia Polytechnic Institute and State University), Dr. Nicolaas Baudoin and Dr. Daniela Cimini (Department of Biological Sciences, Virginia Polytechnic Institute and State University), and Tsubasa Toda and Akari Ueta (Department of Environmental and Life Sciences, Toyohashi University of Technology) for technical assistance. We also thank all the past an current members of the Green, Takahashi, and Kojima laboratories for invaluable discussion. This work was supported by the Luther and Alice Hamlett Undergraduate Research Support (to C.T.S), Howard Hughes Medical Institute (to J.S.T), and National Institutes of Health GM127122 (to C.B.G), and GM126223 (to S.K.). J.S.T. is an Investigator in the Howard Hughes Medical Institute.
Publisher Copyright:
© 2021 Mosig et al.
PY - 2021/6/1
Y1 - 2021/6/1
N2 - In mammals, a set of core clock genes form transcription–translation feedback loops to generate circadian oscillations. We and others recently identified a novel transcript at the Period2 (Per2) locus that is transcribed from the antisense strand of Per2. This transcript, Per2AS, is expressed rhythmically and antiphasic to Per2 mRNA, leading to our hypothesis that Per2AS and Per2 mutually inhibit each other’s expression and form a double negative feedback loop. By perturbing the expression of Per2AS, we found that Per2AS transcription, but not transcript, represses Per2. However, Per2 does not repress Per2AS, as Per2 knockdown led to a decrease in the Per2AS level, indicating that Per2AS forms a single negative feedback loop with Per2 and maintains the level of Per2 within the oscillatory range. Per2AS also regulates the amplitude of the circadian clock, and this function cannot be solely explained through its interaction with Per2, as Per2 knockdown does not recapitulate the phenotypes of Per2AS perturbation. Overall, our data indicate that Per2AS is an important regulatory molecule in the mammalian circadian clock machinery. Our work also supports the idea that antisense transcripts of core clock genes constitute a common feature of circadian clocks, as they are found in other organisms.
AB - In mammals, a set of core clock genes form transcription–translation feedback loops to generate circadian oscillations. We and others recently identified a novel transcript at the Period2 (Per2) locus that is transcribed from the antisense strand of Per2. This transcript, Per2AS, is expressed rhythmically and antiphasic to Per2 mRNA, leading to our hypothesis that Per2AS and Per2 mutually inhibit each other’s expression and form a double negative feedback loop. By perturbing the expression of Per2AS, we found that Per2AS transcription, but not transcript, represses Per2. However, Per2 does not repress Per2AS, as Per2 knockdown led to a decrease in the Per2AS level, indicating that Per2AS forms a single negative feedback loop with Per2 and maintains the level of Per2 within the oscillatory range. Per2AS also regulates the amplitude of the circadian clock, and this function cannot be solely explained through its interaction with Per2, as Per2 knockdown does not recapitulate the phenotypes of Per2AS perturbation. Overall, our data indicate that Per2AS is an important regulatory molecule in the mammalian circadian clock machinery. Our work also supports the idea that antisense transcripts of core clock genes constitute a common feature of circadian clocks, as they are found in other organisms.
KW - Amplitude
KW - Antisense transcript
KW - Circadian
KW - Long noncoding RNA]
KW - Period2
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U2 - 10.1101/gad.343541.120
DO - 10.1101/gad.343541.120
M3 - Article
C2 - 34016691
AN - SCOPUS:85107366582
VL - 38
JO - Genes and Development
JF - Genes and Development
SN - 0890-9369
IS - 11-12
ER -