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Journal of Virology, October 2005, p. 13037-13046, Vol. 79, No. 20
0022-538X/05/$08.00+0     doi:10.1128/JVI.79.20.13037-13046.2005
Copyright © 2005, American Society for Microbiology. All Rights Reserved.

Human Herpesvirus 6 Open Reading Frame U14 Protein and Cellular p53 Interact with Each Other and Are Contained in the Virion

Department of Microbiology, Osaka University Graduate School of Medicine C1, 2-2 Yamada-Oka Suita, Osaka 565-0871, Japan,¹ Department of Cell Biology and Neuroscience, Osaka University Graduate School of Medicine A1, 2-2 Yamada-Oka Suita, Osaka 565-0871, Japan,² Department of Microbiology, Jikei University School of Medicine, Minato-ku, Tokyo 105-8461, Japan³

Received 5 January 2005/ Accepted 23 June 2005

A mass spectroscopic analysis of proteins from human herpesvirus 6 (HHV-6)-infected cells showed that the HHV-6 U14 protein coimmunoprecipitated with the tumor suppressor p53. The binding of U14 to p53 was verified by coimmunoprecipitation experiments in both Molt-3 cells infected with HHV-6 and 293 cells cotransfected with U14 and p53 expression vectors. Indirect immunofluorescence assays (IFAs) showed that by 18 h postinfection (hpi) U14 localized to the dot-like structures observed in both the nucleus and cytoplasm where p53 was partly accumulated. Despite Northern blotting evidence that U14 follows late kinetics, the U14 protein was detected immediately after infection (at 3 hpi) by IFA. In addition, by Western blotting, U14 was detected at 0 hpi or in the presence of cycloheximide which completely abolished the expression of IE1 protein. In addition to U14, p53 was detected at 0 hpi although it was not detected in mock-infected cells. Furthermore, both U14 and p53 were clearly detected in the viral particles by Western blotting and immunoelectron microscopy, supporting the idea that U14 and p53 are incorporated into virions. Our study provides the first evidence of the incorporation of cellular p53 into viral particles and suggests that p53 may play an important role in viral infection.

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