Mammalian circadian rhythm-like gene

#1. Field of the Invention

#The present invention relates generally to the fields of molecular genetics and neuroendocrine cellular biology. More specifically, the present invention relates to a mammalian drosophila period-like gene that exhibits properties of a circadian pacemaker.

#2. Description of the Related Art

#The file of this patent contains at least one drawing executed in color. Copies of this patent with color drawing(s) will be provided by the Patent and Trademark Office upon request and payment of the necessary fee.

#So that the matter in which the above-recited features, advantages and objects of the invention, as well as others which will become clear, are attained and can be understood in detail, more particular descriptions of the invention briefly summarized above may be had by reference to certain embodiments thereof which are illustrated in the appended drawings. These drawings form a part of the specification. It is to be noted, however, that the appended drawings illustrate preferred embodiments of the invention and therefore are not to be considered limiting in their scope.

#FIGS. 1A-1D show the characterization of the human RIGUI gene. FIG. 1A shows the fluorescence in situ hybridization (FISH) mapping of a cosmid 39C2 (LA17NC01) containing the RIGUI gene to human metaphase chromosomes. The gene maps is located on the short arm of chromosome 17. FIG. 1B shows the structure of three RIGUI isoforms deduced from extensive screening of a human heart cDNA library. The three isoforms were identical in regions indicated by the same hue/shading. The proteins diverge in their C-terminal portion. The blue box indicates a putative basic-helix-loop-helix motif and the pink box represents the PAS domain with the two characteristic repeats A and B. Thick lines in RIGUI 6.6 3′ untranslated regions (UTR) represent sequences not present in the RIGUI 3.0 and RIGUI 4.7 variants. FIG. 1C shows a Northern blot of human heart muscle RNA probed with the RIGUI cDNA. Note the transcripts at 4.7 and 6.6 kb, possibly representing RIGUI 4.7 and RIGUI 6.6 cDNAs shown in FIG. 1B. FIG. 1D shows the Northern analysis of adult of adult mouse tissues. Transcript sizes (arrows) are similar to those seen in humans.

#The present invention describes the identification of a putative mammalian per ortholog designated as RIGUI (named after an ancient Chinese sundial). In the mouse, this gene exhibits striking circadian changes of expression in the retina, the suprachiasmatic nucleus, the pars tuberalis of the median eminence, and in the Purkinje cells of the cerebellum. As expected for a pacemaker gene, RIGUI oscillation is maintained under constant darkness. The cycle of oscillation can be entrained upon shifting the light/dark setting. Remarkably, RIGUI expression in the pars tuberalis, the retina, the Purkinje cells and suprachiasmatic nucleus is not in phase, indicating that expression in these tissues oscillates in a tissue-autonomous fashion. Taken together, the present invention suggest that RIGUI is a mammalian ortholog of the Drosophila period gene, and that a key regulator of circadian rhythms is thus conserved during evolution.

#In accordance with the present invention there may be employed conventional molecular biology, microbiology, and recombinant DNA techniques within the skill of the art. Such techniques are explained fully in the literature. See, e.g., Maniatis, Fritsch & Sambrook, “Molecular Cloning: A Laboratory Manual” (1982); “DNA Cloning: A Practical Approach,” Volumes I and II (D. N. Glover ed. 1985); “Oligonucleotide Synthesis” (M. J. Gait ed. 1984); “Nucleic Acid Hybridization” [B. D. Hames & S. J. Higgins eds. (1985)]; “Transcription and Translation” [B. D. Hames & S. J. Higgins eds. (1984)]; “Animal Cell Culture” [R. I. Freshney, ed. (1986)]; “Immobilized Cells And Enzymes” [IRL Press, (1986)]; B. Perbal, “A Practical Guide To Molecular Cloning” (1984). Therefore, if appearing herein, the following terms shall have the definitions set out below.

#The amino acid described herein are preferred to be in the “L” isomeric form. However, residues in the “D” isomeric form can be substituted for any L-amino acid residue, as long as the desired functional property of immunoglobulin-binding is retained by the polypeptide. NH2 refers to the free amino group present at the amino terminus of a polypeptide. COOH refers to the free carboxy group present at the carboxy terminus of a polypeptide. In keeping with standard polypeptide nomeclature, abbreviations for amino acid residues are shown below: