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Nucleic Acids Research, 1989, Vol. 17, No. 8 3261-3273
© 1989

MOLECULAR BIOLOGY

Sequences that promote formation of catenated intertwines during termination of DNA replication

Shawn C. Field-Berry+ and Melvin L. DePamphilis*,§

Department of Biological Chemistry, Harvard Medical School Boston, MA 02115. USA

*To whom correspondence should be addressed

Received September 19, 1988. Revised March 8, 1989. Accepted March 8, 1989.

The normal sequence at which SV40 DNA replication terminates (TER) is unusual in that it promotes formation of catenated intertwines when two converging replication forks enter to complete replication (Weaver et al., 1985). Here we show that yeast centromeric sequences also exhibit this phenomenon. CEN3 caused accumulation of late replicating intermediates and catenated dimers in plasmids replicating in mammalian cells, but only when it was located in the termination region (180° from ori), and only when cells were subjected to hypertonic shock to reduce topoisomerase II activity. Therefore, formation of catenated intertwines during termination of DNA replication was sequence dependent, suggesting that topoisomerase II acts behind replication forks in the termination region to remove intertwines generated by unwinding DNA rather than acting after replication is completed and catenates are formed. Under normal physiological conditions, CEN3 did not promote formation of catenated dimers in either mammalian or yeast cells. Therefore, CEN does not maintain association of sister chromatids during mitosis in yeast by introducing stable catenated intertwines during replication.

+Present addresses: Department of Genetics, Harvard Medical School, 25 Shattuck Street, Boston, MA 02115, USA

§Department of Cell and Developmental Biology, Roche Institute of Molecular Biology, Roche Research Center, Nutley, NJ 07110, USA


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