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The Journal of Cell Biology, Vol 56, 230-245, Copyright © 1973 by Rockefeller University Press

ARTICLE

SINGLE STRAND-CONTAINING REPLICATING MOLECULES OF CIRCULAR MITOCHONDRIAL DNA

David R. Wolstenholme 1, Katsuro Koike 1, and Patricia Cochran-Fouts 1

1 From the Division of Biology, Kansas State University, Manhattan, Kansas 66502 and the Department of Biology, University of Utah, Salt Lake City, Utah 84112

The present address of Dr. Wolstenholme and Mrs. Cochran-Fouts is the latter Dr. Koike's present address is the Laboratory of Biophysics, Institute of Medical Sciences, University of Tokyo, Tokyo, Japan.

Mitochondrial DNAs (mtDNAs) from Chang rat solid hepatomas and Novikoff rat ascites hepatomas were examined in the electron microscope after preparation by the aqueous and by the formamide protein monolayer techniques. MtDNAs from both tumors were found to include double-forked circular molecules with a form and size suggesting they were replicative intermediates. These molecules were of two classes. In molecules of one class, all three segments were apparently totally double stranded. Molecules of the second class were distinguished by the fact that one of the segments spanning the region between the forks in which replication had occurred (the daughter segments) was either totally single stranded, or contained a single-stranded region associated with one of the forks. Daughter segments of both totally double-stranded and single strand-containing replicating molecules varied in length from about 3 to about 80% of the circular contour length of the molecule. Similar classes of replicating molecules were found in mtDNA from regenerating rat liver and chick embryos, indicating them to be normal intermediates in the replication of mtDNA All of the mtDNAs examined included partially single-stranded simple (nonforked) circular molecules. A possible scheme for the replication of mtDNA is presented, based on the different molecular forms observed

Submitted on March 21, 1972
Revised on August 28, 1972


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