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Molecular and Cellular Biology, October 2008, p. 6278-6289, Vol. 28, No. 20
0270-7306/08/$08.00+0     doi:10.1128/MCB.02242-07
Copyright © 2008, American Society for Microbiology. All Rights Reserved.

Drosophila ATF-2 Regulates Sleep and Locomotor Activity in Pacemaker Neurons{triangledown}

Hideyuki Shimizu,1,{dagger} Masami Shimoda,2,{dagger} Terumi Yamaguchi,2 Ki-Hyeon Seong,1 Tomoo Okamura,1,3 and Shunsuke Ishii1,3*

Laboratory of Molecular Genetics, RIKEN Tsukuba Institute, Tsukuba, Ibaraki 305-0074, Japan,1 National Institute of Agrobiological Sciences, Tsukuba, Ibaraki 305-8634, Japan,2 University of Tsukuba, Graduate School of Comprehensive Human Sciences, Tsukuba, Ibaraki 305-8577, Japan3

Received 19 December 2007/ Returned for modification 2 February 2008/ Accepted 30 July 2008

Stress-activated protein kinases such as p38 regulate the activity of transcription factor ATF-2. However, the physiological role of ATF-2, especially in the brain, is unknown. Here, we found that Drosophila melanogaster ATF-2 (dATF-2) is expressed in large ventral lateral neurons (l-LNvs) and also, to a much lesser extent, in small ventral lateral neurons, the pacemaker neurons. Only l-LNvs were stained with the antibody that specifically recognizes phosphorylated dATF-2, suggesting that dATF-2 is activated specifically in l-LNvs. The knockdown of dATF-2 in pacemaker neurons using RNA interference decreased sleep time, whereas the ectopic expression of dATF-2 increased sleep time. dATF-2 knockdown decreased the length of sleep bouts but not the number of bouts. The ATF-2 level also affected the sleep rebound after sleep deprivation and the arousal threshold. dATF-2 negatively regulated locomotor activity, although it did not affect the circadian locomotor rhythm. The degree of dATF-2 phosphorylation was greater in the morning than at night and was enhanced by forced locomotion via the dp38 pathway. Thus, dATF-2 is activated by the locomotor while it increases sleep, suggesting a role for dATF-2 as a regulator to connect sleep with locomotion.

* Corresponding author. Mailing address: Laboratory of Molecular Genetics, RIKEN Tsukuba Institute, Tsukuba, Ibaraki 305-0074, Japan. Phone: 81-29-836-9031. Fax: 81-29-836-9030. E-mail: sishii{at}rtc.riken.jp

{triangledown} Published ahead of print on 11 August 2008.

{dagger} H.S. and M.S. contributed equally to the work.

Molecular and Cellular Biology, October 2008, p. 6278-6289, Vol. 28, No. 20
0270-7306/08/$08.00+0     doi:10.1128/MCB.02242-07
Copyright © 2008, American Society for Microbiology. All Rights Reserved.



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