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J. Biol. Chem., Vol. 275, Issue 47, 36671-36675, November 24, 2000

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NF-B Regulates Transcription of the Mouse Telomerase Catalytic Subunit^*

Lei Yin, Andrea K. Hubbard^§, and Charles Giardina^¶

From the  Department of Molecular and Cell Biology and the ^§ Department of Pharmaceutical Sciences, University of Connecticut, Storrs, Connecticut 06269

Expression of the telomerase catalytic subunit (TERT) is the rate-limiting determinant of telomerase activity in most cells. Analysis of the mouse TERT promoter revealed a potential NF-B binding site 350 base pairs upstream from the translational start site. An oligonucleotide from this region of the TERT promoter bound to proteins in a nuclear extract prepared from a mouse hepatoma cell line. These proteins were identified as NF-B by a number of criteria: 1) the protein complex formed on the TERT oligonucleotide had an electrophoretic mobility similar to that formed on an NF-B consensus oligonucleotide; 2) protein binding to this site was enhanced by NF-B activators tumor necrosis factor-, phorbol 12-myristate 13-acetate, and interleukin-1; and 3) the complex was specific and could be supershifted with antibodies against the p50 or p65 NF-B subunits. The NF-B binding site from the mouse TERT promoter activated transcription when fused to a basal SV40 promoter and enhanced the activity of the native TERT promoter in mouse hepatoma cells stimulated with phorbol 12-myristate 13-acetate. Transcriptional activation by the TERT NF-B site could also be enhanced by co-transfection with an NF-B1 expression vector. NF-B may therefore contribute to the activation of TERT expression observed in mouse tissue.

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