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Journal of Biochemistry Advance Access published online on October 3, 2008
Journal of Biochemistry, doi:10.1093/jb/mvn129
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© 2008 The Japanese Biochemical Society

Identification and characterization of a positive regulatory cis-element within the upstream region of c-jun

Shivani Agarwal, Rajkumar*, Preeti Gupta and Aparna Dixit{dagger}

Gene Regulation Laboratory, School of Biotechnology, Jawaharlal Nehru University, New Delhi-110067, India

{dagger} To whom correspondence should be addressed. Professor Aparna Dixit, Gene Regulation Laboratory, School of Biotechnology, Jawaharlal Nehru University, New Delhi-110 067, India, Tel No: +91-11-26742164; +91-11-26704085, Fax No: +91-11-26717561, E-mail: adix2100{at}mail.jnu.ac.in; adixit7{at}yahoo.com, adixit7{at}hotmail.com

Received July 9, 2008; Accepted September 15, 2008


  Abstract

Jun oncoprotein, a component of transcription factor AP-1, is responsible for expression of multiple genes which trigger cell proliferation, differentiation and apoptosis. The diverse regulatory roles that c-Jun plays point towards its varied and complex transcriptional regulation under different physiological conditions. This could possibly involve an interplay of different trans-acting factors and multiple cis-acting elements present in the c-jun. It is, therefore, important to identify such regulatory elements that could be involved in its stringent up- or down-regulation. In the present study, transient transfection analysis with stepwise deletion variants of the upstream region (-563 to -273), linked to basal c-jun promoter in a reporter plasmid revealed the existence of multiple regulatory regions that modulated gene expression. Further delineation of the region, conferring positive regulation of transcription from the c-jun promoter, led to the identification of a cis-acting element spanning -538 to -514 region. The trans-acting factor(s) present in rat liver nuclear extract interacted specifically only in the phosphorylated form, and with a high affinity to the recognition sequence. UV-cross linking and South-Western blotting, in conjunction with the analysis of affinity purified proteins interacting with this region, revealed that at least two protein factors of (45 kDa and (34 kDa bind specifically to this region.

Key Words: c-jun, cis-acting element, EMSA, Positive regulatory factor, transcriptional regulation

*Present address: Division of Health and Reproductive Sciences, Lexington, Kentuky-40202, USA


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