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Nucleic Acids Research, 1981, Vol. 9, No. 21 5697-5710
© 1981

MOLECULAR BIOLOGY

The high affinity binding site on polyoma virus DNA for the viral large-T protein

Patrick Gaudray*, Chiara Tyndall+, Robert Kamen+ and Francois Cuzin*

*Centre de Biochimie du CNRS, Université de Nice 06034 Nice, France +Transcription Laboratory, Imperial Cancer Research Fund Laboratories P.O. Box 123, Lincoln's Inn Fields, London, UK

Received August 27, 1981. In order to map the high affinity binding site for the viral large-Tprotein on polyoma virus DNA, we have developped an assay which does not require purified protein. It is based on the specific elution of the large-TATPase activity from calf thynus DNA cellulose by reccmbinant DNA moleculesincluding known sequences of the viral DNA. Using this assay, a high affinity binding site has been mapped on the early region side of the ori region. Binding requires the integrity of a sequence /AGAGGC/TTCC/AGAGGC7 Tnucleotides 49 to 64 in the DNA sequence of the A2 strain). Similar repeats of aPuGFuGGC sequence within less than 20 bases are not found within the viralcoding regions, but are strikingly common in the control regions of papova-virusesand other eukaryotic DNAs.


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