The structure of replicating kinetoplast DNA networks

David Pérez-Morga, Paul T. Englund

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Abstract

Kinetoplast DNA (kDNA), the mitochondrial DNA of Crithidia fasciculata and related trypanosomatids, is a network containing ∼5,000 covalently closed minicircles which are topologically interlocked. kDNA synthesis involves release of covalently closed minicircles from the network, and, after replication of the free minicircles, reattachment of the nicked or gapped progeny minicircles to the network periphery. We have investigated this process by electron microscopy of networks at different stages of replication. The distribution of nicked and closed minicircles is easily detectable either by autoradiography of networks radiolabeled at endogenous nicks by nick translation or by twisting the covalently closed minicircles with intercalating dye. The location of newly synthesized minicircles within the network is determined by autoradiography of networks labeled in vivo with a pulse of [3H]thymidine. These studies have clarified structural changes in the network during replication, the timing of repair of nicked minicircles after replication, and the mechanism of division of the network.

Original languageEnglish (US)
Pages (from-to)1069-1079
Number of pages11
JournalJournal of Cell Biology
Volume123
Issue number5
Publication statusPublished - Dec 1993

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ASJC Scopus subject areas

  • Cell Biology

Cite this

Pérez-Morga, D., & Englund, P. T. (1993). The structure of replicating kinetoplast DNA networks. Journal of Cell Biology, 123(5), 1069-1079.