Mammalian TIMELESS is involved in period determination and DNA damage-dependent phase advancing of the circadian clock.

The transcription/translation feedback loop-based molecular oscillator underlying the generation of circadian gene expression is preserved in almost all organisms. Interestingly, the animal circadian clock proteins CRYPTOCHROME (CRY), PERIOD (PER) and TIMELESS (TIM) are strongly conserved at the ami...

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Autores principales:	Erik Engelen, Roel C Janssens, Kazuhiro Yagita, Veronique A J Smits, Gijsbertus T J van der Horst, Filippo Tamanini
Formato:	article
Lenguaje:	EN
Publicado:	Public Library of Science (PLoS) 2013
Materias:	Medicine R Science Q
Acceso en línea:	https://doaj.org/article/c3526d46ec8d42e2a4920d56d7cdf7dd
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spelling	oai:doaj.org-article:c3526d46ec8d42e2a4920d56d7cdf7dd2021-11-18T07:57:36ZMammalian TIMELESS is involved in period determination and DNA damage-dependent phase advancing of the circadian clock.1932-620310.1371/journal.pone.0056623https://doaj.org/article/c3526d46ec8d42e2a4920d56d7cdf7dd2013-01-01T00:00:00Zhttps://www.ncbi.nlm.nih.gov/pmc/articles/pmid/23418588/?tool=EBIhttps://doaj.org/toc/1932-6203The transcription/translation feedback loop-based molecular oscillator underlying the generation of circadian gene expression is preserved in almost all organisms. Interestingly, the animal circadian clock proteins CRYPTOCHROME (CRY), PERIOD (PER) and TIMELESS (TIM) are strongly conserved at the amino acid level through evolution. Within this evolutionary frame, TIM represents a fascinating puzzle. While Drosophila contains two paralogs, dTIM and dTIM2, acting in clock/photoreception and chromosome integrity/photoreception respectively, mammals contain only one TIM homolog. Whereas TIM has been shown to regulate replication termination and cell cycle progression, its functional link to the circadian clock is under debate. Here we show that RNAi-mediated knockdown of TIM in NIH3T3 and U2OS cells shortens the period by 1 hour and diminishes DNA damage-dependent phase advancing. Furthermore, we reveal that the N-terminus of TIM is sufficient for interaction with CRY1 and CHK1 as well for homodimerization, and the C-terminus is necessary for nuclear localization. Interestingly, the long TIM isoform (l-TIM), but not the short (s-TIM), interacts with CRY1 and both proteins can reciprocally regulate their nuclear translocation in transiently transfected COS7 cells. Finally, we demonstrate that co-expression of PER2 abolishes the formation of the TIM/CRY1 complex through affinity binding competition to the C-terminal tail of CRY1. Notably, the presence of the latter protein region evolutionarily and structurally distinguishes mammalian from insect CRYs. We propose that the dynamic interaction between these three proteins could represent a post-translational aspect of the mammalian circadian clock that is important for its pace and adaption to external stimuli, such as DNA damage and/or light.Erik EngelenRoel C JanssensKazuhiro YagitaVeronique A J SmitsGijsbertus T J van der HorstFilippo TamaniniPublic Library of Science (PLoS)articleMedicineRScienceQENPLoS ONE, Vol 8, Iss 2, p e56623 (2013)
institution	DOAJ
collection	DOAJ
language	EN
topic	Medicine R Science Q
spellingShingle	Medicine R Science Q Erik Engelen Roel C Janssens Kazuhiro Yagita Veronique A J Smits Gijsbertus T J van der Horst Filippo Tamanini Mammalian TIMELESS is involved in period determination and DNA damage-dependent phase advancing of the circadian clock.
description	The transcription/translation feedback loop-based molecular oscillator underlying the generation of circadian gene expression is preserved in almost all organisms. Interestingly, the animal circadian clock proteins CRYPTOCHROME (CRY), PERIOD (PER) and TIMELESS (TIM) are strongly conserved at the amino acid level through evolution. Within this evolutionary frame, TIM represents a fascinating puzzle. While Drosophila contains two paralogs, dTIM and dTIM2, acting in clock/photoreception and chromosome integrity/photoreception respectively, mammals contain only one TIM homolog. Whereas TIM has been shown to regulate replication termination and cell cycle progression, its functional link to the circadian clock is under debate. Here we show that RNAi-mediated knockdown of TIM in NIH3T3 and U2OS cells shortens the period by 1 hour and diminishes DNA damage-dependent phase advancing. Furthermore, we reveal that the N-terminus of TIM is sufficient for interaction with CRY1 and CHK1 as well for homodimerization, and the C-terminus is necessary for nuclear localization. Interestingly, the long TIM isoform (l-TIM), but not the short (s-TIM), interacts with CRY1 and both proteins can reciprocally regulate their nuclear translocation in transiently transfected COS7 cells. Finally, we demonstrate that co-expression of PER2 abolishes the formation of the TIM/CRY1 complex through affinity binding competition to the C-terminal tail of CRY1. Notably, the presence of the latter protein region evolutionarily and structurally distinguishes mammalian from insect CRYs. We propose that the dynamic interaction between these three proteins could represent a post-translational aspect of the mammalian circadian clock that is important for its pace and adaption to external stimuli, such as DNA damage and/or light.
format	article
author	Erik Engelen Roel C Janssens Kazuhiro Yagita Veronique A J Smits Gijsbertus T J van der Horst Filippo Tamanini
author_facet	Erik Engelen Roel C Janssens Kazuhiro Yagita Veronique A J Smits Gijsbertus T J van der Horst Filippo Tamanini
author_sort	Erik Engelen
title	Mammalian TIMELESS is involved in period determination and DNA damage-dependent phase advancing of the circadian clock.
title_short	Mammalian TIMELESS is involved in period determination and DNA damage-dependent phase advancing of the circadian clock.
title_full	Mammalian TIMELESS is involved in period determination and DNA damage-dependent phase advancing of the circadian clock.
title_fullStr	Mammalian TIMELESS is involved in period determination and DNA damage-dependent phase advancing of the circadian clock.
title_full_unstemmed	Mammalian TIMELESS is involved in period determination and DNA damage-dependent phase advancing of the circadian clock.
title_sort	mammalian timeless is involved in period determination and dna damage-dependent phase advancing of the circadian clock.
publisher	Public Library of Science (PLoS)
publishDate	2013
url	https://doaj.org/article/c3526d46ec8d42e2a4920d56d7cdf7dd
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Mammalian TIMELESS is involved in period determination and DNA damage-dependent phase advancing of the circadian clock.

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