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Originally published In Press as doi:10.1074/jbc.M011541200 on June 29, 2001

J. Biol. Chem., Vol. 276, Issue 35, 32489-32494, August 31, 2001
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An Abd-B Class HOX·PBX Recognition Sequence Is Required for Expression from the Mouse Ren-1c Gene*

Li PanDagger , Youhua Xie§, Thomas A. Black, Craig A. Jones, Steven C. Pruitt, and Kenneth W. Gross||

From the Department of Molecular and Cellular Biology, Roswell Park Cancer Institute, Buffalo, New York 14263

Expression from the mouse Ren-1c gene in As4.1 cells is dependent on a proximal promoter element (PPE) located at approximately -60 and a 241-base pair enhancer region located at -2625 relative to the transcription start site. The PPE (TAATAAATCAA) is identical to a consensus HOX·PBX binding sequence. Further, PBX1b has been shown to be a component of a PPE-specific binding complex present in nuclear extracts from As4.1 cells. The binding affinities of different paralog HOX members to the PPE were examined in the absence or presence of PBX1b. HOXB6, -B7, and -C8 failed to bind the PPE alone but showed weak affinity in the presence of PBX1b. In contrast, HOXD10 and to a lesser degree HOXB9 bound the PPE with high affinities regardless of whether PBX1b was present. Abd-B HOX members, including HOXD10, -A10, -A9, -B9, and -C9, are expressed in As4.1 cells. The ability of HOX and PBX1b to form a ternary complex with PREP1 on the PPE is also demonstrated both in vivo and in vitro. Point mutations in either the HOX or PBX half-site of the PPE disrupted the formation of the HOX·PBX complex and dramatically decreased transcriptional activity of the Ren-1c gene demonstrating that both the HOX and PBX half-sites are critical for mouse renin gene expression. These results strongly implicate Abd-B class Hox genes and their cofactors as major determinants of the sites of renin expression.

* This work was supported by National Institutes of Health Grants HL48459 (to K. W. G.), CA16056, and HD36416 (to S. C. P.).The costs of publication of this article were defrayed in part by the payment of page charges. The article must therefore be hereby marked "advertisement" in accordance with 18 U.S.C. Section 1734 solely to indicate this fact.

This manuscript is offered belatedly in honor of Prof. Max L. Birnstiel's retirement as Director of the Institute of Molecular Pathology, Vienna, Austria.

Dagger Supported by a postdoctoral fellowship from the National Institutes of Health.

§ Present address: Mondogen GmbH, Am Klopferspitz 19a, D-82152 Martinsried, Germany.

Present address: School of Medicine and Biomedical Sciences, 40 CFS Bldg., 3435 Main St., Buffalo, NY 14214.

|| To whom correspondence should be addressed: Dept. of Molecular and Cellular Biology, Roswell Park Cancer Institute, Elm and Carlton Sts., Buffalo, NY 14263-0001. Tel.: 716-845-4572; Fax: 716-845-8169; E-mail: gross@acsu.buffalo.edu.

Copyright © 2001 by The American Society for Biochemistry and Molecular Biology, Inc.


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