Specific binding of single-stranded telomeric DNA by Cdc13p of Saccharomyces cerevisiae

The Journal of Biological Chemistry
Y C LinJ J Lin

Abstract

Cdc13p is a single strand telomere-binding protein of Saccharomyces cerevisiae; its telomere-binding region is within amino acids 451-693, Cdc13(451-693)p. In this study, we used purified Cdc13p and Cdc13(451-693)p to characterize their telomere binding activity. We found that the binding specificity of single-stranded TG(1-3) DNA by these two proteins is similar. However, the affinity of Cdc13(451-693)p to DNA was slightly lower than that of Cdc13p. The binding of telomeric DNA by these two proteins was disrupted at NaCl concentrations higher than 0.3 m, indicating that electrostatic interaction contributed significantly to the binding process. Because both proteins bound to strand TG(1-3) DNA positioned at the 3' end, the 5' end, or in the middle of the oligonucleotide substrates, our results indicated that the location of TG(1-3) in single-stranded DNA does not appear to be important for Cdc13p binding. Moreover, using DNase I footprint analysis, the structure of the telomeric DNA complexes of Cdc13p and Cdc13(451-693)p was analyzed. The DNase I footprints of these two proteins to three different telomeric DNA substrates were virtually identical, indicating that the telomere contact region of Cdc13p is within Cdc13(451-693)p...Continue Reading

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Citations

Oct 5, 2011·Proceedings of the National Academy of Sciences of the United States of America·Yun Wu, Virginia A Zakian
Mar 12, 2009·The Journal of Biological Chemistry·Tzung-Ju WuJing-Jer Lin
Oct 4, 2006·Journal of the Royal Society, Interface·C J ProctorT B L Kirkwood
Oct 5, 2001·Molecular and Cellular Biology·K Förstemann, J Lingner
Jun 26, 2010·Protein Expression and Purification·Wei QianJin-Qiu Zhou
Dec 2, 2005·Journal of Molecular Biology·J RhodinM Cohn
Sep 24, 2016·Scientific Reports·Yu-Fan ChenJing-Jer Lin
Aug 11, 2017·Journal of Cell Science·Hsi-Hsien HsiehJing-Jer Lin
Oct 5, 2001·The Journal of Biological Chemistry·Y C LinJ J Lin
Jan 10, 2003·The Journal of Biological Chemistry·Te-Ling PangJing-Jer Lin
Sep 7, 2007·The Biochemical Journal·Yi-Ching HsiehJing-Jer Lin
Apr 9, 2021·Journal of the American Chemical Society·Yi-Yun LinJing-Jer Lin

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