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Papers In Press, published online ahead of print January 10, 2002
Physiology and Cell Biology, The Ohio State University College of Medicine and Public Health, Columbus, OH 43210
Corresponding Author: lee.2076{at}osu.edu
The proton-translocating vacuolar ATPase (V-ATPase) acidifies the endocytic network of eukaryotic cells. Although all eukaryotic cell types require low to moderate levels of V-ATPase, some proton-secreting cells express amplified levels for use in specialized membrane domains. To characterize genetic elements required for this heightened expression, we studied transcription and stability of mRNA encoding the V-ATPase c subunit in a low-expressing fibroblast cell line (NIH 3T3), and a high-expressing macrophage cell line (RAW 264.7). Isolation of the promoter and mapping of the transcriptional start site indicated that the c subunit promoter is TATA-less and initiates transcription at a single site. Promoter activity was regulated through the same transcription factor binding sites in both cell types, which showed no discernible difference in rates of c subunit transcription. In contrast, c subunit transcripts showed markedly greater stability in RAW cells than in 3T3 cells, as did other constitutively expressed V-ATPase subunit transcripts. Only the B and ‘a’ subunits, which are expressed in multiple isoforms, were not regulated solely by mRNA stability. These results suggest that overall expression levels of the V-ATPase are set primarily by regulation of mRNA stability, and that transcriptional mechanisms determine subunit composition in varying cell types.
J. Biol. Chem, 10.1074/jbc.M111959200
Submitted on December 14, 2001
Revised on January 10, 2002
Accepted on January 9, 2002
Regulation of enhanced vacuolar H+-ATPase expression in macrophages
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