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. 2010 Nov 24;39(7):2580–2592. doi: 10.1093/nar/gkq1222

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© The Author(s) 2010. Published by Oxford University Press.

This is an Open Access article distributed under the terms of the Creative Commons Attribution Non-Commercial License (http://creativecommons.org/licenses/by-nc/2.5), which permits unrestricted non-commercial use, distribution, and reproduction in any medium, provided the original work is properly cited.

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Figure 4. — Sld2T84D binds to ssARS1-1. (A) Biotinylated dsARS1, ssARS1-2 or ssARS1-1 (7 pmol) was conjugated to streptavidin agarose and then mixed with varying amounts of radiolabeled Sld2T84D, as indicated in the figure, for 5 min at 30°C. As a control, beads without DNA were incubated with Sld2T84D. The beads were washed and analyzed as described in ‘Materials and Methods’ section. The results from experiments similar to (A) were quantified and plotted as the percent Sld2T84D bound versus the input protein concentration (B). Sld2T84D binds more tightly to ssARS1-1 than either ssARS1-2 or dsARS1. (C) Biotinylated ssARS1-1 was tested for binding to CDK-phosphorylated Sld2 and PKA-phosphorylated Sld2. The results from experiments similar to (C) were quantified and plotted as the fraction of Sld2 bound versus the input protein concentration (D). These results suggest CDK phosphorylation of Sld2 stimulates binding to ssARS1-1.