The double-time protein kinase regulates the subcellular localization of the Drosophila clock protein period.
The Period (PER), Timeless (TIM), and Double-Time (DBT) proteins are essential components of one feedback loop in the Drosophila circadian molecular clock. PER and TIM physically interact. Coexpression of PER and TIM promotes their nuclear accumulation and influences the activity of DBT: although DBT phosphorylates and destabilizes PER, this is suppressed by TIM. Experiments using Drosophila cells in culture have indicated that PER can translocate to the nucleus without TIM and will repress transcription in a DBT-potentiated manner. In this study, we examined the control of PER subcellular localization in Drosophila clock cells in vivo. We found that PER can translocate to the nucleus in tim(01) null mutants but only if DBT kinase activity is inhibited. We also found that nuclear PER is a potent transcriptional repressor in dbt mutants in vivo without TIM. Thus, in vivo, DBT regulates PER subcellular localization, in addition to its previously documented role as a mediator of PER stability. However, DBT does not seem essential for transcriptional repression by PER. It was reported previously that overexpression of a second kinase, Shaggy (SGG)/Glycogen Synthase Kinase 3, accelerates PER nuclear accumulation. Here, we show that these effects of SGG on PER nuclear accumulation require TIM. We propose a revised clock model that incorporates this tight kinase regulation of PER and TIM nuclear entry.
Cyran
Shawn A
SA
Department of Biology, New York University, New York, New York 10003, USA.
Yiannoulos
Georgia
G
Buchsbaum
Anna M
AM
Saez
Lino
L
Young
Michael W
MW
Blau
Justin
J
D056930:ARNTL Transcription Factors; D021581:Active Transport, Cell Nucleus; D000818:Animals; D051792:Basic Helix-Loop-Helix Proteins / Q000037:antagonists & inhibitors / Q000378:metabolism; D056926:CLOCK Proteins; D048149:Casein Kinase 1 epsilon / Q000037:antagonists & inhibitors / Q000378:metabolism / Q000502:physiology*; D002467:Cell Nucleus / Q000378:metabolism; D002940:Circadian Rhythm; D004330:Drosophila / Q000235:genetics / Q000378:metabolism*; D029721:Drosophila Proteins / Q000037:antagonists & inhibitors / Q000235:genetics / Q000378:metabolism / Q000502:physiology*; D038362:Glycogen Synthase Kinase 3 / Q000502:physiology; D007150:Immunohistochemistry; D008954:Models, Biological; D009154:Mutation; D009687:Nuclear Proteins / Q000378:metabolism*; D056950:Period Circadian Proteins; D014157:Transcription Factors / Q000037:antagonists & inhibitors / Q000378:metabolism; D014158:Transcription, Genetic
D056930:ARNTL Transcription Factors; D051792:Basic Helix-Loop-Helix Proteins; D056926:CLOCK Proteins; D048149:Casein Kinase 1 epsilon; D029721:Drosophila Proteins; D038362:Glycogen Synthase Kinase 3; D009687:Nuclear Proteins; D056950:Period Circadian Proteins; D014157:Transcription Factors; C120526:Clk protein, Drosophila; C539040:PER protein, Drosophila; C112805:cyc protein, Drosophila; C113519:dco protein, Drosophila; C096445:tim protein, Drosophila; C080335:Sgg protein, Drosophila