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Nucleic Acids Research, 1988, Vol. 16, No. 21 10119-10129
© 1988

ENZYMOLOGY

Effects of VM26, a specific inhibitor of type II DNA topoisomerase, on SV40 chromatin replication in vitro

Arndt Richter and Ulrich Strausfeld

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

Received August 5, 1988. Accepted October 13, 1988.

We have examined the influence of VM26 (teniposide), a specific inhibitor of mammalian type II DNA topoisomerase, on the replication of SV40 minichromosomes in vitro. The replication system we used consists of replicative intermediate SV40 chromatin as substrate which is converted to mature SV40 chromatin in the presence of ATP, deoxynucleotides and a protein extract from uninfected cells. The addition of 100µM VM26 to this system reduces DNA synthesis to 70 to 80 percent of the control and leads to an accumulation of ‘late replicative intermediates’. The VM26 induced block of replication was not released by the addition of large quantities of type I DNA topoisomerase. We conclude, that type II DNA topoisomerase is essential for the final replication steps leading from late Cairns structures of replicative intermediates to monomeric minichromosoms. It appears that type I DNA topoisomerase can function as a swivelase during most of the replicative elongation phase, but must later be replaced by type II DNA topoisomerase.


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