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Kinetic analysis of Sp1-mediated transcriptional activation of the human DNA polymerase β promoter

Oncogene volume 19, pages 4729–4735 (2000)Cite this article

Abstract

In the present studies, we have examined the effect of Sp1 on the activation of the human DNA polymerase β (β-pol), a TATA-less promoter. A HeLa cell nuclear extract (NE) based in vitro runoff transcription system of core β-pol promoter human DNA (pβP8) three-step kinetic model of transcription initiation were used to describe the kinetic effect of Sp1. The results showed that distal Sp1-binding sites in the core β-pol promoter are important for transcriptional activation of the pβP8 promoter. A detailed kinetic analysis showed that Sp1 stimulates the activity of the pβP8 promoter through distal Sp1-binding sites by increasing the amount of recruitment, instead of stimulating the apparent rate of RPc assembly (k1). There was no significant effect of Sp1 on the apparent rate of open complex (RPo) formation (k2) or on the apparent rate of promoter clearance (k3) of the pβP8 promoter. These studies define the kinetic mechanisms by which Sp1 may regulate the rate of transcript formation of the pβP8 promoter, and these results may have implications for Sp1 regulation of TATA-less promoters.

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Acknowledgements

This work was supported by NIH grants CA77721 (to S Narayan) and ES06492 (to SH Wilson).

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Authors and Affiliations

  1. Sealy Center for Molecular Science, University of Texas Medical Branch, Galveston, 77555, Texas, TX, USA

    Satya Narayan

  2. Sealy Center for Oncology and Hematology, University of Texas Medical Branch, Galveston, 77555, Texas, TX, USA

    Satya Narayan

  3. Laboratory of Structural Biology, National Institutes of Environmental Health Sciences, National Institutes of Health, Research Triangle Park, North Carolina, 27709, NC, USA

    Samuel H Wilson

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Narayan, S., Wilson, S. Kinetic analysis of Sp1-mediated transcriptional activation of the human DNA polymerase β promoter. Oncogene 19, 4729–4735 (2000). https://doi.org/10.1038/sj.onc.1203823

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