TY - JOUR
T1 - Molecular cloning and characterization of the human CLOCK gene
T2 - Expression in the suprachiasmatic nuclei
AU - Steeves, Thomas D L
AU - King, David P.
AU - Zhao, Yaliang
AU - Sangoram, Ashvin M.
AU - Du, Fenghe
AU - Bowcock, Anne M.
AU - Moore, Robert Y.
AU - Takahashi, Joseph S.
N1 - Funding Information:
We thank Andrew Whiteley and Lisa Wilsbacher for assistance with sequencing; Anne Phung, To Hao Thai, and Elizabeth Yetman for technical help with exon determination; and Philip Lowrey and Marina Antoch for valuable discussions on subcloning protocols. This work was supported by the NSF Center for Biological Timing, United States Air Force Office of Scientific Research Grant 97NC170, an Unrestricted Grant in Neuroscience from Bristol-Myers Squibb, and NIMH Grant R37MH39592. J.S.T. is an Investigator in the Howard Hughes Medical Institute.
PY - 1999/4/15
Y1 - 1999/4/15
N2 - The Clock gene is an essential regulator of circadian rhythms. It encodes a member of the basic helix-loop-helix/PER-ARNT-SIM family of transcription factors known to play a central role in the control of diverse cellular events. Previously we described the functional identification and molecular isolation of the Clock gene in the mouse, its interaction with the BMAL1 protein, and the role of this complex as a transcriptional activator in the circadian pacemaker. Here, we report the cloning, exon organization, chromosomal location, and mRNA expression of the human CLOCK gene. The coding sequence of human CLOCK extends for 2538 bp and is 89% identical to its mouse ortholog; its deduced amino acid sequence is 846 residues long and is 96% identical to mouse CLOCK. Radiation hybrid mapping localized human CLOCK to the long arm of human chromosome 4 (4q12). Direct sequencing of a genomic CLOCK clone indicated that the coding sequence of human CLOCK extends over 20 exons and that its intron/exon organization is identical to that of the mouse ortholog. Northern blot analysis indicated widespread expression of two major transcripts of 8 and 10 kb, and in situ hybridization of human brain tissue revealed elevated expression of CLOCK mRNA in the suprachiasmatic nuclei, the locus of circadian control in mammals, and in the cerebellum. Comparison of cDNA clones revealed two single nucleotide polymorphisms in noncoding sequence flanking the CLOCK open reading frame. The central role of Clock in the organization of circadian rhythms suggests that it will be a useful candidate gene for genetic analyses of disorders associated with dysfunction of the circadian system.
AB - The Clock gene is an essential regulator of circadian rhythms. It encodes a member of the basic helix-loop-helix/PER-ARNT-SIM family of transcription factors known to play a central role in the control of diverse cellular events. Previously we described the functional identification and molecular isolation of the Clock gene in the mouse, its interaction with the BMAL1 protein, and the role of this complex as a transcriptional activator in the circadian pacemaker. Here, we report the cloning, exon organization, chromosomal location, and mRNA expression of the human CLOCK gene. The coding sequence of human CLOCK extends for 2538 bp and is 89% identical to its mouse ortholog; its deduced amino acid sequence is 846 residues long and is 96% identical to mouse CLOCK. Radiation hybrid mapping localized human CLOCK to the long arm of human chromosome 4 (4q12). Direct sequencing of a genomic CLOCK clone indicated that the coding sequence of human CLOCK extends over 20 exons and that its intron/exon organization is identical to that of the mouse ortholog. Northern blot analysis indicated widespread expression of two major transcripts of 8 and 10 kb, and in situ hybridization of human brain tissue revealed elevated expression of CLOCK mRNA in the suprachiasmatic nuclei, the locus of circadian control in mammals, and in the cerebellum. Comparison of cDNA clones revealed two single nucleotide polymorphisms in noncoding sequence flanking the CLOCK open reading frame. The central role of Clock in the organization of circadian rhythms suggests that it will be a useful candidate gene for genetic analyses of disorders associated with dysfunction of the circadian system.
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U2 - 10.1006/geno.1998.5675
DO - 10.1006/geno.1998.5675
M3 - Article
C2 - 10198158
AN - SCOPUS:0344483899
SN - 0888-7543
VL - 57
SP - 189
EP - 200
JO - Genomics
JF - Genomics
IS - 2
ER -