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Kusano, Y.

Publications and source records attributed to Kusano, Y..

2 recordsLinked to original sources

Progressive chromosome shape changes during cell divisions, recapitulated by loop capture simulations

Mitotic chromosomes give genome portions the required compaction and mechanical stability for faithful inheritance during cell divisions. Here, we record human chromosome dimensions from their appearance in prophase over successive times in a mitotic arrest. Chromosomes first appear long and uniformly thin. Then, individual chromosome arms become discernible, which continuously shorten and thicken - the longer a chromosome arm, the thicker it becomes. The observed chromosome arm length to width relationship can be described by a power law with progressively increasing exponent. In the search for a molecular explanation of this behavior, the popular loop extrusion model provides no obvious means by which longer arms become thicker. Instead, we find that simulations of an alternative loop capture model recapitulate key features of our observations, including the gradually developing arm length to width relationship. Our analyses portray chromosomes as out-of-equilibrium structures in the process of transitioning towards, but on biologically relevant time scales not typically reaching, steady state.

biophysics↗

Spatial organization of supercoil dynamics during DNA replication

Progression of DNA replication inevitably generates helical tension and resulting super-helical structures, or supercoils, that originate from the double helix of DNA. As DNA polymerases progress, negative supercoils accumulate in the wake of the replication fork, thereby impeding fork progression unless the supercoils are resolved topologically. Using super-resolution microscopy combined with spatial distribution analysis, here we describe that SMC5/6-mediated confinement of negative supercoils lies at the basis of releasing them by topoisomerase Top2A. Along with DNA replication, SMC5/6 progressively associates with chromatin and locally increases the density of supercoil clusters behind the fork, allowing Top2A to efficiently target and release the accumulated supercoils. These processes are essential to complete DNA replication prior to mitosis and therefore to ensure genome stability. Remarkably, we found that HeLa cells over-accumulate poorly confined negative supercoils beyond the processing capacity of cellular SMC5/6, which may exemplify the condition associated with genome instability of cancer cells.

cell biology↗