THE TOPOLOGY OF THE KINETOPLAST DNA NETWORK

Kinetoplast DNA (kDNA) of trypanosomatid parasites is a network of similar to 5000 catenated DNA minicircles and similar to 25 maxicircles. We developed the following strategy to deduce the topological linkage of the minicircles of the Crithidia fasciculata network. First, we used graph theory to provide precise models of possible network structures. Second, on the basis of these models, we predicted the frequencies of minicircle oligomers expected from random network breakage. Third, we determined the fragmentation pattern of kDNA networks as a function of the extent of digestion. Fourth, by comparison of the results with the predictions, we identified the model that best represents the network. We conclude that each minicircle is linked on average to three other minicircles. A honeycomb arrangement probably results, with each minicircle typically at the vertex of a hexagonal cell. This topology has implications for the assembly, structure, and function of kDNA networks.