Transactivation mechanisms of mouse clock transcription factors, mClock and mArnt3.
BACKGROUND
The Arnt3 (also termed as BMAL1 or MOP3)/Clock heterodimer is a positive regulator of circadian rhythm and activates the transcription of target genes such as per1 and vasopressin.


RESULTS
We investigated the transcriptional mechanism of mArnt3/mClock heterodimer. While mClock did not possess any distinct activation domain, mArnt3 contained a transcriptional activation domain at the most C-terminal end, the activity of which was not expressed, even in the one hybrid system, until it was bound by mClock. It has been suggested that mClock plays a regulatory or structural role in exerting a transcription enhancing effect of the mArnt3/mClock heterodimer. Deletion proceeding from amino acids 559-492 of mClock markedly reduced the transactivation activity of mArnt3/mClock heterodimer, in consistence with the results of the Clock-delta 19 mutant. Yeast and mammalian two-hybrid systems revealed that CBP and p300 interacted with mArnt3 via the CREB binding domain. The In vivo interaction between mArnt3 and CBP was confirmed by the GST pull down assay.


CONCLUSIONS
Taken together, these results suggest that the mArnt3/mClock heterodimer exerted its transactivation activity via CBP or p300 interacting with mArnt3 in the heterodimer with mClock playing a structural or regulatory role in the transactivation process.
Takahata
S
S
Department of Chemistry, Graduate School of Science, Tohoku University, Aoba-ku, Sendai 980-8578, Japan.
Ozaki
T
T
Mimura
J
J
Kikuchi
Y
Y
Sogawa
K
K
Fujii-Kuriyama
Y
Y
D056930:ARNTL Transcription Factors; D000818:Animals; D051784:Aryl Hydrocarbon Receptor Nuclear Translocator; D056926:CLOCK Proteins; D050882:CREB-Binding Protein; D002352:Carrier Proteins / Q000235:genetics / Q000378:metabolism*; D002460:Cell Line; D002940:Circadian Rhythm; D050881:E1A-Associated p300 Protein; D017930:Genes, Reporter; D015151:Immunoblotting; D008156:Luciferases / Q000235:genetics / Q000378:metabolism; D051379:Mice; D009687:Nuclear Proteins / Q000378:metabolism; D011485:Protein Binding; D017434:Protein Structure, Tertiary; D015534:Trans-Activators / Q000235:genetics / Q000378:metabolism*; D015533:Transcriptional Activation; D020798:Two-Hybrid System Techniques
D056930:ARNTL Transcription Factors; C538961:Bmal1 protein, mouse; D002352:Carrier Proteins; D009687:Nuclear Proteins; D015534:Trans-Activators; D051784:Aryl Hydrocarbon Receptor Nuclear Translocator; D008156:Luciferases; D056926:CLOCK Proteins; D050882:CREB-Binding Protein; C538944:Clock protein, mouse; C494364:Crebbp protein, mouse; D050881:E1A-Associated p300 Protein; C494361:Ep300 protein, mouse