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Molecular and Cellular Biology, August 2006, p. 6094-6104, Vol. 26, No. 16
0270-7306/06/$08.00+0     doi:10.1128/MCB.02366-05
Copyright © 2006, American Society for Microbiology. All Rights Reserved.

Transcriptional Pausing Caused by NELF Plays a Dual Role in Regulating Immediate-Early Expression of the junB Gene

Masatoshi Aida,1 Yexi Chen,1 Koichi Nakajima,2 Yuki Yamaguchi,1 Tadashi Wada,1 and Hiroshi Handa1*

Graduate School of Bioscience and Biotechnology, Tokyo Institute of Technology, Midori-ku, Yokohama 226-8503, Japan,1 Graduate School of Medicine, Osaka City University, Abeno-ku, Osaka 545-8585, Japan2

Received 12 December 2005/ Returned for modification 4 January 2006/ Accepted 30 May 2006

Human 5,6-dichloro-1-ß-D-ribofuranosylbenzimidazole sensitivity-inducing factor (DSIF) and negative elongation factor (NELF) negatively regulate transcription elongation by RNA polymerase II (RNAPII) in vitro. However, the physiological roles of this negative regulation are not well understood. Here, by using a number of approaches to identify protein-DNA interactions in vivo, we show that DSIF- and NELF-mediated transcriptional pausing has a dual function in regulating immediate-early expression of the human junB gene. Before induction by interleukin-6, RNAPII, DSIF, and NELF accumulate in the promoter-proximal region of junB, mainly at around position +50 from the transcription initiation site. After induction, the association of these proteins with the promoter-proximal region continues whereas RNAPII and DSIF are also found in the downstream regions. Depletion of a subunit of NELF by RNA interference enhances the junB mRNA level both before and after induction, indicating that DSIF- and NELF-mediated pausing contributes to the negative regulation of junB expression, not only by inducing RNAPII pausing before induction but also by attenuating transcription after induction. These regulatory mechanisms appear to be conserved in other immediate-early genes as well.

* Corresponding author. Mailing address: Graduate School of Bioscience and Biotechnology, Tokyo Institute of Technology, Midori-ku, Yokohama 226-8503, Japan. Phone: 81-45-924-5872. Fax: 81-45-924-5834. E-mail: hhanda{at}bio.titech.ac.jp.

Molecular and Cellular Biology, August 2006, p. 6094-6104, Vol. 26, No. 16
0270-7306/06/$08.00+0     doi:10.1128/MCB.02366-05
Copyright © 2006, American Society for Microbiology. All Rights Reserved.



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