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The Journal of Neuroscience, February 1, 1999, 19(3):987-994

The 69 bp Circadian Regulatory Sequence (CRS) Mediates per-Like Developmental, Spatial, and Circadian Expression and Behavioral Rescue in Drosophila

Haiping Hao^{1, 2, 3}, Nick R. J. Glossop¹, Lisa Lyons¹, Jan Qiu^{1, 2, 4}, Bronwyn Morrish^{1, 5}, Yuzhong Cheng^{1, 2, 6}, Charlotte Helfrich-Förster⁷, and Paul Hardin¹

¹ Department of Biology, University of Houston, Houston, Texas 77204-5513, ² Department of Biology, Texas A & M University, College Station, Texas 77843-3258, ³ Department of Pediatrics, Yale University School of Medicine, New Haven, Connecticut 06520-8081, ⁴ Department of Biology, Brandeis University, Waltham, Massachusetts 02254, ⁵ Department of Biology, University of York, Heslington YO1 5DD, United Kingdom, ⁶ Department of Cell Biology, Baylor College of Medicine, Houston, Texas 77030, and ⁷ Institute of Zoology, Animal Physiology, University of Tübingen, Tübingen 72076, Germany

The period (per) gene is an essential component of the circadian timekeeping mechanism in Drosophila. This gene is expressed in a circadian manner, giving rise to a protein that feeds-back to regulate its own transcription. A 69 bp clock regulatory sequence (CRS) has been identified previously upstream of the period gene. The CRS confers wild-type mRNA cycling when used to drive a lacZ reporter gene in transgenic flies. To determine whether the CRS also mediates proper developmental and spatial expression and behavioral rescue, we used the CRS to drive either lacZ or per in transgenic flies. The results show that the CRS is able to activate expression in pacemaker neuron precursors in larvae and essentially all tissues that normally express per in pupae and adults. The CRS is sufficient to rescue circadian feedback loop function and behavioral rhythms in per⁰¹ flies. However, the period of locomotor activity rhythms shortens if a stronger basal promoter is used. This study shows that regulatory elements sufficient for clock-dependent and tissue-specific per expression in larvae, pupae, and adults are present in the CRS and that the period of adult locomotor activity rhythms is dependent, in part, on the overall level of per transcripts.

Key words: Drosophila; circadian clock; transcriptional regulation; behavior; period gene; developmental expression

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Copyright © 1999 Society for Neuroscience  0270-6474/99/193987-08$05.00/0

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