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Volume 272, Number 1, Issue of January 3, 1997 pp. 174-181
©1997 by The American Society for Biochemistry and Molecular Biology, Inc.

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A Novel Transcription Factor Regulates Expression of the Vacuolar H⁺-ATPase B2 Subunit through AP-2 Sites during Monocytic Differentiation

(Received for publication, August 6, 1996, and in revised form, October 21, 1996)

Beth S. Lee , Irina Krits , Monica K. Crane-Zelkovic and Stephen L. Gluck ^¶

From the  Department of Medicine/Renal Division, ^¶ Department of Cell Biology and Physiology, and the  George M. O'Brien Center for Kidney and Urological Diseases, Washington University School of Medicine, St. Louis, Missouri 63110

During monocyte-to-macrophage differentiation, the cellular content of vacuolar H⁺-ATPase (V-ATPase) increases more than 4-fold. We have shown previously that amplified expression of the B2 subunit of the V-ATPase occurs solely by increased transcription, and that the 5-untranslated region of the B2 gene, containing multiple consensus binding sites for the transcription factors AP-2 and Sp1, is required for this expression. The present study demonstrates that AP-2 binding sequences are essential for increased transcription from the B2 promoter during monocyte-macrophage differentiation and that AP-2, expressed exogenously in THP-1 and other cells, activates transcription from the B2 promoter. In mobility shift assays, a nuclear factor from THP-1 and U-937 cells was identified that binds to several AP-2 response elements within the B2 promoter, but does not react with AP-2 antibodies, and has a DNA sequence binding affinity profile that differs from AP-2. These findings suggest that a novel AP-2-like transcription factor is responsible for V-ATPase B subunit amplification during monocyte differentiation.

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