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Oct-1 cooperates with the TATA binding initiation complex to control rapid transcription of human iNOS

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Abstract

Expression of the human inducible nitric oxide synthase (hiNOS) is generally undetectable in resting cells, but stimulation by a variety of signals including cytokines induces transcription in most cell types. The tight transcriptional regulation of the enzyme is a complex mechanism many aspects of which remain unknown. Here, we describe an octamer (Oct) element in hiNOS proximal promoter, located close to the TATA box. This site constitutively binds Oct-1 and its deletion abrogates cytokine-induced transcription, showing that it is indispensable though not sufficient for transcription. Increasing the distance between Oct and the TATA box by inserting inert DNA sequence inhibits transcription, and footprinting of this region shows no other protein binding in resting cells, suggesting an interaction between the two complexes. Chromatin immunoprecipitation assays detect the presence of Oct-1, RNA polymerase II and trimethyl K4 histone H3 on the proximal promoter in resting cells, confirming that the gene is primed for transcription before stimulation. RT-PCR of various fragments along the hiNOS gene shows that transcription is initiated in resting cells and this is inhibited by interference with Oct-1 binding to the proximal site of the promoter. We propose that, through interaction with the initiation complex, Oct-1 regulates hiNOS transcription by priming the gene for the rapid response required in an immune response.

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Acknowledgments

This project was supported by the Ligue Bourgignonne Contre Le Cancer. A.P. was funded by the Fondation pour la Recherche Medicale and an Individual Marie Curie Fellowship from the European Commission. S.R. was supported by the Conseil Regional de Bourgogne. The authors are grateful to Ben Luisi and Tony Jackson for helpful scientific discussions, Fatima Bentrari for technical assistance and Julian Rayner for his support for the completion of this work.

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Author notes

  1. Christopher E. Goldring

    Present address: Department of Pharmacology and Therapeutics, School of Biomedical Sciences, University of Liverpool, Liverpool, UK

  2. Aurélie Chantôme

    Present address: Nutrition, Criossance et Cancer, University of Tours, Tours, France

Authors and Affiliations

  1. EPHE Laboratory, Faculty of Medicine, University of Bourgogne, 7 Bvd Jeanne D’Arc, 21033, Dijon, France

    Sylvie Reveneau, Christopher E. Goldring, Aurélie Chantôme, Jean-François Jeannin & Alena Pance

  2. The Wellcome Trust Sanger Institute, Genome Campus, Hinxton, Cambridge, CB10 1SA, UK

    Thodoris G. Petrakis & Alena Pance

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  1. Sylvie Reveneau
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Correspondence to Alena Pance.

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Reveneau, S., Petrakis, T.G., Goldring, C.E. et al. Oct-1 cooperates with the TATA binding initiation complex to control rapid transcription of human iNOS. Cell. Mol. Life Sci. 69, 2609–2619 (2012). https://doi.org/10.1007/s00018-012-0939-z

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  • DOI: https://doi.org/10.1007/s00018-012-0939-z

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