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Nucleic Acids Research, 1990, Vol. 18, No. 9 2715-2720
© 1990

CHEMISTRY

Loss of DNA-binding and new transcriptional trans activation function in polymavirus large T-antigen with mulation of zinc finger motif

Anders Bergvist, Mats Nilsson, Kåre Bondeson and Göran Magnu•on*

Department of Medical Virology, Uppsala University Biomedical Center Box 584, S-751 23 Uppsala, Sweden

*To whom correspondence should be addressed

Received January 3, 1990. Accepted April 9, 1990.

A putative zinc finger in polyomavirus large T-antigen was Investigated. We were unable to demonstrate unequivocally a requirement for zinc in specific DNAbinding using the chelating agent 1,10-phenarrthroline. An involvement of the putative zinc finger in specific DNA-binding was nevertheless suggested by the properties of a mutant protein with a cysser replacement in the finger motif. Probably as a result of the defective DNA-binding, the mutant protein had lost its activity in initiation of viral DNA-replication and in negative regulation of viral early transcription. However, the frans-activation of the viral late promoter was normal. The analysis also revealed a previously unrecognized activity of large T-antigen. The mutant protein frans-activated the viral early promoter. In the wild-type protein this activity is probably concealed by the separate, negative regulatory function.


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