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Definition of a major p53 binding site on Ad2E1B58K protein and a possible nuclear localization signal on the Ad12E1B54K protein

Oncogene volume 18, pages 955–965 (1999)Cite this article

Abstract

Previous studies have established that adenovirus 2/5 early region 1B (Ad E1B) 58K protein binds p53 strongly and co-localizes with it to cytoplasmic dense bodies whilst the homologous Ad12E1B54K protein binds only weakly and co-localizes primarily to the nucleus in Ad12E1 transformed cells. We have used these properties of the E1B proteins from different viral serotypes to map the p53 binding site on the Ad2/5 protein. A set of chimaeric genes was constructed containing different proportions of the Ad12 and Ad2E1B DNA. These, together with Ad12E1A and E1B19K DNA, were transfected into baby rat kidney cells and transformed lines isolated. From an examination of the properties of these Ad12/Ad2E1B fusion proteins in co-immunoprecipitation and subcellular localization experiments it has been concluded that the p53 binding site on Ad2E1B58K protein lies between amino acids 216 and 235 and that the homologous region on Ad12E1B54K protein also binds p53. In addition, a unique nuclear localization signal is located on Ad12E1B54K between residues 228 and 239. We suggest that primary structure differences in these regions of the Ad2 and Ad12E1B proteins are responsible for the different subcellular localizations in AdE1 transformants.

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Acknowledgements

We are most grateful to Dr Chris Dawson (Institute for Cancer Studies, University of Birmingham) for helpful discussion, Katja Schmeiser for production of figures, Nicola Waldron for invaluable secretarial assistance and the Cancer Research Campaign, UK for financial support.

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  1. Roger JA Grand and Julian Parkhill: RJA Grand and J Parkhill contributed equally to this work.

Authors and Affiliations

  1. CRC Institute for Cancer Studies, University of Birmingham, Edgbaston, Birmingham, B15 2TA, UK

    Roger JA Grand, Julian Parkhill, Tadge Szestak, Susan M Rookes, Sally Roberts & Phillip H Gallimore

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  1. Roger JA Grand
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Grand, R., Parkhill, J., Szestak, T. et al. Definition of a major p53 binding site on Ad2E1B58K protein and a possible nuclear localization signal on the Ad12E1B54K protein. Oncogene 18, 955–965 (1999). https://doi.org/10.1038/sj.onc.1202358

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