Authors:

Gabriel A. Kenning

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About Paper:

Structural Maintenance of Chromosome (SMC) proteins are essential in the regulation of chromosome structure and gene expression in eukaryotic cells. Some Archaea, like those of the Sulfolobus family, contain chromosomal features that show similarity to those of eukaryotic cells. SMC-like proteins help regulate the Sulfolobus chromosome, including Coalescin, or ClsN. Coalescin has been observed to form loops and condense Chromatin, helping produce A and B compartments. It has also been shown that Sulfolobus uses CIsN to achieve this. This series of experiments seeks to better understand how CISsN is activated to regulate chromosomal structure within Sulfolobus. Being a thermophilic and acidophilic organism, Sulfolobus, and therefore CIsN, is predicted to behave optimally at low pH and high temperature. The objective is to determine the critical force - the force that the protein produces - under acidic (pH 5.2) and high temperature (60 C) conditions. This is accomplished through the use of magnetic tweezers. A magnetic bead is attached to one end of a DNA strand, and the other end of the DNA strand is attached to the cover slip of a flow cell. A variable magnetic field is applied, and the extension of the DNA strand is observed through a microscope. Variance in DNA extension when introduced to ClsN and a ClsN-ATP mixture helps determine the force with which ClsN compresses and organizes DNA. The findings of this experiment are that Acidic conditions can somewhat regularly aid in the compression of DNA by as much as 10% at 0.7pN, and that high-temperature conditions without lowered pH do not allow CIsN to compress DNA.

Source:

Northwestern University

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Co-authors:

Gabriel A. Kenning