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Nucleic Acids Research, 1993, Vol. 21, No. 18 4231-4238
© 1993

MOLECULAR BIOLOGY

Roles of novobiocin-sensitive topoisomerases in chloroplast DNA replication in Chlamydomonas reinhardtii

Mark A. Woelfle, Robert J. Thompson* and Gisela Mosig

Department of Molecular Biology, Vanderbilt University Nashville, TN 37235, USA

*To whom correspondence should be addressed

Received June 8, 1993. Revised August 2, 1993. Accepted August 2, 1993.

We have examined DNA replication In Chlamydomonas reinhardtii chloroplasts in vivo when chloroplast type II topoisomerases are inactivated with sublethal doses of novoblocln. DNA replication Is at first inhibited under these conditions. However, after a delay of several hours, chloroplast chromosomes Initiate a novobiocin-Insensitive mode of DNA replication. This replication starts preferentially near a hotspot of recombination in the large inverted repeats, Instead of from the normal chloroplast origins, orlA and oriB. It replicates one, but not the other single-copy region of the chloroplast chromosome. We speculate that novoblocln-insensltive DNA replication In chloroplasts requires recombination in this preferred Initiation region.


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