Interactions between N- and C-terminal domains of the Saccharomyces cerevisiae high-mobility group protein HMO1 are required for DNA bending

Biochemistry
Kevin T BauerleAnne Grove

Abstract

The Saccharomyces cerevisiae high-mobility group protein HMO1 is composed of two DNA-binding domains termed box A and box B, of which only box B is predicted to adopt a HMG fold, and a lysine-rich C-terminal extension. To assess the interaction between individual domains and their contribution to DNA binding, several HMO1 variants were analyzed. Using circular dichroism spectroscopy, thermal stability was measured. While the melting temperatures of HMO1-boxA and HMO1-boxB are 57.2 and 47.2 degrees C, respectively, HMO1-boxBC, containing box B and the entire C-terminal tail, melts at 46.1 degrees C, suggesting little interaction between box B and the tail. In contrast, full-length HMO1 exhibits a single melting transition at 47.9 degrees C, indicating that interaction between box A and either box B or the tail destabilizes this domain. As HMO1-boxAB, lacking only the lysine-rich C-terminal segment, exhibits two melting transitions at 46.0 and 63.3 degrees C, we conclude that the destabilization of the box A domain seen in full-length HMO1 is due primarily to its interaction with the lysine-rich tail. Determination of DNA substrate specificity using electrophoretic mobility shift assays shows unexpectedly that the lysine-rich tai...Continue Reading

References

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Citations

Feb 15, 2012·Biochemistry·Sreerupa Ray, Anne Grove
Jul 25, 2007·Molecular and Cellular Biology·Koji KasaharaTetsuro Kokubo
Sep 19, 2007·Molecular and Cellular Biology·Axel B BergerOlivier Gadal
Nov 19, 2013·Analytical Biochemistry·Qinqin ZhuangChristopher M Wood
Jun 28, 2014·Biochimica Et Biophysica Acta·Matias I HeppJosé L Gutiérrez
Apr 1, 2016·Epigenetics & Chromatin·Arvind Panday, Anne Grove
Nov 21, 2014·Bioscience, Biotechnology, and Biochemistry·Ayako HigashinoKoji Kasahara
Sep 1, 2015·World Journal of Biological Chemistry·Xin Bi
Dec 3, 2016·Microbiology and Molecular Biology Reviews : MMBR·Arvind Panday, Anne Grove
Nov 20, 2016·Genes to Cells : Devoted to Molecular & Cellular Mechanisms·Koji KasaharaTetsuro Kokubo
Mar 18, 2017·Biophysics Reviews·Divakaran MurugesapillaiMark C Williams

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