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The Kinetoplast DNA (kDNA) is a two-dimensional Olympic-ring-like network ofmutually linked 2.5 kb-long DNA minicircles found in certain parasites calledTrypanosomes. Understanding the self-assembly and replication of this structureare not only major open questions in biology but can also inform the design ofsynthetic topological materials. Here we report the first high-resolution,single-molecule study of kDNA network topology using AFM and steered moleculardynamics simulations. We map out the DNA density within the network and thedistribution of linking number and valence of the minicircles. We alsocharacterise the DNA hubs that surround the network and show that they cause abuckling transition akin to that of a 2D elastic thermal sheet in the bulk.Intriguingly, we observe a broad distribution of density and valence of theminicircles, indicating heterogeneous network structure and individualism ofdifferent kDNA structures. Our findings explain outstanding questions in thefield and offer single-molecule insights into the properties of a uniquetopological material.

Single-Molecule Structure and Topology of Kinetoplast DNA Networks