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Nucleic Acids Research, 1989, Vol. 17, No. 1 93-106
© 1989

MOLECULAR BIOLOGY

Sequence independent duplex DNA opening reaction catalysed by SV40 large tumor antigen

Martin Scheffner, Rainer Wessel and Hans Stahl

Fakultät für Biologie, Universität Konstanz D-7750 Konstanz, FRG

Received October 20, 1988. Revised December 2, 1988. Accepted December 2, 1988.

Simian virus 40 (SV40) large tumor antigen (T antigen) is mainly localized in the nucleus where it exhibits two biochemical properties: DNA binding and helicase activity. Both activities are necessary for viral DNA replication and may also enable T antigen to modulate cellular growth. Here we present biochemical and electron microscopic evidence that the helicase activity can start at internal sites of fully double-stranded DNA molecules not containing the SV40 origin of replication. Using T antigen specific monoclonal antibodies, this unwinding reaction can be biochemically divided in an initiation (duplex opening) and a propagation step. The duplex opening reaction (as well as the propagation step) does not depend on a specific DNA sequence or secondary structure. In addition, we have found that T antigen forms an ATP dependent nucleoprotein complex at double-stranded DNA, which may be an essential step for the sequence independent duplex DNA opening reaction.


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