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Nucleic Acids Research, 1993, Vol. 21, No. 24 5570-5576
© 1993

MOLECULAR BIOLOGY

Specific transcriptional activation in vitro by the herpes simplex virus protein VP16

David N. Arnosti+, Chris M. Preston1, Michael Hagmann, Walter Schaffner, R.Graham Hope1, Gerard Laughlan1 and Ben F. Luisi1,*

Institut fur Molekularbiologie II, Universitat Zurich Zurich, Switzerland 1MRC Virology Unit Church Street, Glasgow G11 5JR, UK

*To whom correspondence should be addressed

Received September 23, 1993. Revised November 8, 1993. Accepted November 8, 1993.

The herpes simplex virus protein VP16 interacts with cellular factors, including the protein Oct-1, to activate viral immediate early (IE) gene transcription. We have reproduced this effect by addition of purified, fulllength VP16 and the DNA-binding 'POU' domain of Oct-1 (Oct-1&2.urule;POU) to a HeLa cell in vitro transcription system. Stimulation of transcription was dependent on the IE-specific element, TAATGARAT. In agreement with earlier observations from electrophoretic mobility shift assays, activation was not observed when Oct-2&2.urule;POU, the DNA-binding domain from the Oct-2 protein, was substituted for Oct-1&2.urule;POU. Single round transcription assays revealed that, together, VP16 and Oct-1&2.urule;POU facilitate the assembly of pre-initiation complexes at target gene promoters.

+Present address: University of California, San Diego, Department of Biology, 0322, 9500 Gilman Drive, La Jolla, CA 92093-0322, USA


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