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J. Biol. Chem., Vol. 283, Issue 3, 1317-1323, January 18, 2008

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Interplay between E-box and NF-B in Regulation of A20 Gene by DRB Sensitivity-inducing Factor (DSIF)^*

From the Department of Biological Chemistry, The Weizmann Institute of Science, Rehovot 76100, Israel

The NF-B target gene A20 serves as a paradigm for gene-specific control of transcription elongation. This gene is regulated by the elongation factor DSIF (DRB sensitivity-inducing factor) under basal and NF-B-activated states by two distinct mechanisms. Prior to NF-B stimulation, the A20 gene is occupied by polymerase II, and elongation is inhibited by DSIF. This inhibition is mediated by an upstream promoter element termed ELIE (elongation inhibitory element). Upon NF-B activation, inhibition of the A20 gene by DSIF persists, but now NF-B and the core promoter regulate DSIF instead of ELIE. Here we investigated the regulation of DSIF by ELIE and the regulatory switch from ELIE to NF-B following NF-B induction. Electrophoretic mobility shift assays revealed two distinct protein complexes that specifically interact with ELIE, one of which is the E-box protein USF1. Interestingly, USF1 is displaced from the A20 promoter upon induction of NF-B. A mutation in the E-box section of ELIE diminished the binding of USF1 and DSIF recruitment. Consistent with these findings, the E-box is crucial for DSIF inhibition in resting, but not NF-B-stimulated, cells. These findings reveal a dynamic regulation of DSIF involving either E-box or NF-B depending on the physiological circumstances.

Received for publication, August 14, 2007 , and in revised form, October 16, 2007.

^* This work was supported by grants from the Israel Cancer Research Foundation and the Israel Science Foundation. The costs of publication of this article were defrayed in part by the payment of page charges. This article must therefore be hereby marked "advertisement" in accordance with 18 U.S.C. Section 1734 solely to indicate this fact.

¹ To whom correspondence should be addressed. Tel.: 972-8-9342117; Fax: 972-8-9344118; E-mail: rivka.dikstein{at}weizmann.ac.il.

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