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Journal of Virology, February 2008, p. 1908-1922, Vol. 82, No. 4
0022-538X/08/$08.00+0     doi:10.1128/JVI.01716-07
Copyright © 2008, American Society for Microbiology. All Rights Reserved.

Intracellular Localization Map of Human Herpesvirus 8 Proteins{triangledown}

Gaby Sander,1,{dagger} Andreas Konrad,1,{dagger} Mathias Thurau,1 Effi Wies,2 Rene Leubert,3 Elisabeth Kremmer,4 Holger Dinkel,5 Thomas Schulz,6 Frank Neipel,2 and Michael Stürzl1*

Division of Molecular and Experimental Surgery, Department of Surgery, University of Erlangen-Nuremberg, Schwabachanlage 10, D-91054 Erlangen, Germany,1 Institute of Clinical and Molecular Virology, University of Erlangen-Nuremberg, Schlossgarten 4, D-91054 Erlangen, Germany,2 Department of Virus-Induced Vasculopathy, GSF-National Research Center for Environment and Health, Ingolstädter Landstrasse 1, D-85764 Neuherberg, Germany,3 GSF-Service Unit Monoclonal Antibodies and Cell Sorting, GSF-National Research Center for Environment and Health, Marchioninistrasse 25, D-81377 Munich, Germany,4 Division of Bioinformatics, Institute of Biochemistry, University of Erlangen-Nuremberg, Fahrstrasse 17, D-91054 Erlangen, Germany,5 Medical School Hannover, Department of Virology, Carl-Neubergstrasse 1, D-30625 Hannover, Germany6

Received 7 August 2007/ Accepted 21 November 2007

Human herpesvirus 8 (HHV-8) is the etiological agent of Kaposi's sarcoma. We present a localization map of 85 HHV-8-encoded proteins in mammalian cells. Viral open reading frames were cloned with a Myc tag in expression plasmids, confirmed by full-length sequencing, and expressed in HeLa cells. Protein localizations were analyzed by immunofluorescence microscopy. Fifty-one percent of all proteins were localized in the cytoplasm, 22% were in the nucleus, and 27% were found in both compartments. Surprisingly, we detected viral FLIP (v-FLIP) in the nucleus and in the cytoplasm, whereas cellular FLIPs are generally localized exclusively in the cytoplasm. This suggested that v-FLIP may exert additional or alternative functions compared to cellular FLIPs. In addition, it has been shown recently that the K10 protein can bind to at least 15 different HHV-8 proteins. We noticed that K10 and only five of its 15 putative binding factors were localized in the nucleus when the proteins were expressed in HeLa cells individually. Interestingly, in coexpression experiments K10 colocalized with 87% (13 of 15) of its putative binding partners. Colocalization was induced by translocation of either K10 alone or both proteins. These results indicate active intracellular translocation processes in virus-infected cells. Specifically in this framework, the localization map may provide a useful reference to further elucidate the function of HHV-8-encoded genes in human diseases.

* Corresponding author. Mailing address: University of Erlangen-Nuremberg, Department of Surgery, Division of Molecular and Experimental Surgery, Schwabachanlage 10, D-91054 Erlangen, Germany. Phone: 49 9131 85 33109. Fax: 49 9131 85 32077. E-mail: michael.stuerzl{at}uk-erlangen.de

{triangledown} Published ahead of print on 12 December 2007.

{dagger} G.S. and A.K. contributed equally to this study.

Journal of Virology, February 2008, p. 1908-1922, Vol. 82, No. 4
0022-538X/08/$08.00+0     doi:10.1128/JVI.01716-07
Copyright © 2008, American Society for Microbiology. All Rights Reserved.



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