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Nuclear localization of the adenovirus E4orf4 protein is mediated through an arginine-rich motif and correlates with cell death

A Corrigendum to this article was published on 09 June 2005

Abstract

The adenovirus E4orf4 protein induces p53-independent death of human cancer cells by a mechanism requiring interactions with the Bα subunit of protein phosphatase 2A. When expressed alone E4orf4 localizes predominantly in the nucleus, although significant levels are also present in the cytoplasm. While tyrosine phosphorylation of E4orf4 and recruitment of Src have been linked with E4orf4 cytoplasmic cell death functions, little is known about the functions of E4orf4 in the nucleus. In this study, we identified an arginine-rich motif (E4ARM; residues 66–75) that is necessary and sufficient for nuclear and nucleolar localization. This motif, which is highly homologous to the arginine-rich nuclear and nucleolar localization motif of some lentiviral proteins, was shown to target heterologous proteins to the nucleus and to nucleoli, functions found to be dependent on the overall charge of the motif rather than on specific residues. Furthermore, mutation of arginine residues to alanines but not to lysines in E4ARM was shown to block such targeting activity and, when introduced into full-length E4orf4, to decrease induction of cell death. Finally, coexpression of the ARM motifs of E4orf4, HIV-1 Tat or Rev along with full-length E4orf4 was seen to decrease E4orf4-dependent cell killing. Thus it appears that targeting of E4orf4 to the nucleus and cell nucleoli by E4ARM is an important component of E4orf4-induced cell death.

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Abbreviations

ARM:

arginine-rich motif

E4ARM:

adenovirus E4orf4 arginine-rich motif, residues 66–75

HIV-1:

human immunodeficiency virus 1

HTLV-1:

human T-cell leukemia virus 1

NoLS:

nucleolar localization signal

PK:

pyruvate kinase

PP2A:

protein phosphatase 2A

SV40 LT:

simian virus 40 large T antigen

RRNA:

ribosomal RNA

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Acknowledgements

We thank Paola Blanchette and Suiyang Li (McGill University) for helpful discussions and critical review of the manuscript. We are also grateful to Gideon Dreyfuss (University of Pennsylvania, School of Medicine) for the gift of myc-PK cDNA, to John F Presley, Jacynthe Laliberté and Alfredo Ribeiro-DaSilva (McGill University) for assistance with confocal microscopy and to Melissa Mui and Ratna Samanta (McGill University) for making the HA-E4orf4-R66-73K cDNA. This work was supported by grants from the National Cancer Institute of Canada (PEB), the Canadian Institutes of Health Research (PEB, IEG and JNL) and GeminX Biotechnologies Inc. (PEB).

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Correspondence to Philip E Branton.

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Miron, MJ., Gallouzi, IE., Lavoie, J. et al. Nuclear localization of the adenovirus E4orf4 protein is mediated through an arginine-rich motif and correlates with cell death. Oncogene 23, 7458–7468 (2004). https://doi.org/10.1038/sj.onc.1207919

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