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Journal of Virology, September 2004, p. 9512-9523, Vol. 78, No. 17
0022-538X/04/$08.00+0     DOI: 10.1128/JVI.78.17.9512-9523.2004
Copyright © 2004, American Society for Microbiology. All Rights Reserved.

Human Cytomegalovirus Tegument Proteins ppUL82 (pp71) and ppUL35 Interact and Cooperatively Activate the Major Immediate-Early Enhancer

Karina Schierling,1 Thomas Stamminger,2 Thomas Mertens,1 and Michael Winkler1*

Abteilung Virologie, Universitätsklinikum Ulm, Ulm,1 Institut für Klinische und Molekulare Virologie der Universität Erlangen-Nürnberg, Erlangen, Germany2

Received 30 December 2003/ Accepted 27 April 2004

The tegument protein ppUL82 (pp71) of human cytomegalovirus (HCMV) has previously been shown to activate the immediate-early transcription of HCMV and to enhance the infectivity of viral DNA. This is concordant with its localization adjacent to promyelocytic leukemia oncogenic domains (PODs) immediately after infection. In a yeast two-hybrid screen, we identified the tegument protein ppUL35 as an interacting partner of ppUL82. The interaction could be confirmed in transfected and infected cells. The domain responsible for interaction was narrowed down to amino acids 447 to 516 within ppUL35, thus allowing both forms of ppUL35 to interact with ppUL82. Immunofluorescence experiments showed a relocalization of ppUL35 from a diffuse nuclear pattern when expressed alone to PODs when expressed together with ppUL82. In accordance with this observation and the role of ppUL82 as a transactivator, we observed a cooperative activation of the HCMV major immediate-early enhancer but not of heterologous viral enhancer elements. These results suggest an important role for this interaction in the stimulation of viral immediate-early gene expression and viral infection.

* Corresponding author. Mailing address: Abteilung Virologie, Universitätsklinikum Ulm, Albert Einstein Allee 11, D-89081 Ulm, Germany. Phone: 49 731 5002 3336. Fax: 49 731 5002 3337. E-mail: michael.winkler{at}medizin.uni-ulm.de.

Journal of Virology, September 2004, p. 9512-9523, Vol. 78, No. 17
0022-538X/04/$08.00+0     DOI: 10.1128/JVI.78.17.9512-9523.2004
Copyright © 2004, American Society for Microbiology. All Rights Reserved.



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