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Published online 7 May 2001. doi:10.1083/jcb.153.4.735
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© The Rockefeller University Press, 0021-9525/2001/5/735/ $5.00
The Journal of Cell Biology, Volume 153, Number 4, May 14, 2001 735-744


Original Article

Intramitochondrial Location and Dynamics of Crithidia fasciculata Kinetoplast Minicircle Replication Intermediates

Mark E. Drewa and Paul T. Englunda
a Department of Biological Chemistry, Johns Hopkins Medical School, Baltimore, Maryland 21205

Correspondence to: Paul T. Englund, Department of Biological Chemistry, Johns Hopkins Medical School, 725 N. Wolfe St., Baltimore, MD 21205. Tel:(410) 955-3790 Fax:(410) 955-7810 E-mail:penglund{at}jhmi.edu.

Kinetoplast DNA, the mitochondrial DNA of Crithidia fasciculata, is organized into a network containing 5,000 topologically interlocked minicircles. This network, situated within the mitochondrial matrix, is condensed into a disk-shaped structure located near the basal body of the flagellum. Fluorescence in situ hybridization revealed that before their replication, minicircles are released vectorially from the network face nearest the flagellum. Replication initiates in the zone between the flagellar face of the disk and the mitochondrial membrane (we term this region the kinetoflagellar zone [KFZ]). The replicating minicircles then move to two antipodal sites that flank the disk-shaped network. In later stages of replication, the number of free minicircles increases, accumulating transiently in the KFZ. The final replication events, including primer removal, repair of many of the gaps, and reattachment of the progeny minicircles to the network periphery, are thought to take place within the antipodal sites.

Key Words: cell cycle, kinetoplast DNA, DNA replication, fluorescence in situ hybridization, trypanosomatid


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