HOME HELP FEEDBACK SUBSCRIPTIONS ARCHIVE SEARCH TABLE OF CONTENTS		QUICK SEARCH:   [advanced] Author: Keyword(s): Year:  Vol:  Page:

J. Biol. Chem., Vol. 282, Issue 24, 17941-17952, June 15, 2007

This Article Full Text Full Text (PDF) Supplemental Data An addition or correction has been published All Versions of this Article: 282/24/17941    most recent M610618200v1 Submit a Letter to Editor Alert me when this article is cited Alert me when eLetters are posted Alert me if a correction is posted Citation Map Services Email this article to a friend Similar articles in this journal Similar articles in PubMed Alert me to new issues of the journal Download to citation manager Request Permissions Citing Articles Citing Articles via Google Scholar Google Scholar Articles by Ammons, M. C. B. Articles by Gauss, K. A. Search for Related Content PubMed PubMed Citation Articles by Ammons, M. C. B. Articles by Gauss, K. A. Social Bookmarking                      What's this?

Binding of Pleomorphic Adenoma Gene-like 2 to the Tumor Necrosis Factor (TNF)--responsive Region of the NCF2 Promoter Regulates p67^phox Expression and NADPH Oxidase Activity^*

From the Department of Veterinary Molecular Biology, Montana State University, Bozeman, Montana 59717

NCF2, the gene encoding the NADPH oxidase cytosolic component p67^phox, is up-regulated by TNF-, and we recently mapped a region in the NCF2 promoter that was required for this TNF--dependent response. Because this TNF--responsive region (TRR) lacked recognizable transcription factor binding elements, we performed studies to identify factors involved in regulating NCF2 via the TRR. Using the TRR sequence as bait in a yeast one-hybrid screen, we identified the zinc finger transcription factor Pleomorphic Adenoma Gene-Like 2 (PLAGL2) as a candidate regulator of NCF2 expression. PLAGL2-specific antibodies were generated that detected the native and SUMO1-modified forms of endogenous PLAGL2. EMSA and DNA-binding protein affinity purification analyses demonstrated specific binding of in vitro-translated as well as endogenously expressed PLAGL2 to the TRR, and chromatin immunoprecipitation assays demonstrated enhanced binding of endogenous PLAGL2 to the TRR in vivo with TNF- treatment. Knockdown of PLAGL2 protein inhibited up-regulation of NCF2 transcript, p67^phox protein expression, and subsequent superoxide production in response to TNF-. Furthermore, relative levels of native and SUMO1-modified endogenous PLAGL2 protein were modulated in a time-dependant manner in response to TNF- treatment. These data clearly identify PLAGL2 as a novel regulator of NCF2 gene expression as well as NADPH oxidase activity and contribute to a greater understanding of the transcriptional regulation of NCF2.

Received for publication, November 15, 2006 , and in revised form, March 29, 2007.

^* This work was supported in part by National Institutes of Health Grants AR42426 and RR020185, USDA NRI/CGP Grant 2006-01690, an equipment grant from the M. J. Murdock Charitable Trust, and the Montana State University Agricultural Experimental Station. 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.

The on-line version of this article (available at http://www.jbc.org) contains supplemental Figs. S1-S3.

¹ To whom correspondence should be addressed: Veterinary Molecular Biology, Montana State University, Bozeman, MT 59717. Tel.: 406-994-4703; Fax: 406-994-4303; E-mail: kgauss{at}montana.edu.

CiteULike    Complore    Connotea    Del.icio.us    Digg    Reddit    Technorati    What's this?