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. Author manuscript; available in PMC: 2010 Dec 11.
Published in final edited form as: Mol Cell. 2009 Dec 11;36(5):743–753. doi: 10.1016/j.molcel.2009.10.014

Figure 5. A Ddc1 peptide containing the bipartite Mec1 activation sequence activates Mec1.

Figure 5

A 30 mer peptide containing the bipartite Ddc1 sequence from 346 to 365 aa and 537 to 546 aa is shown. Residues Trp352 and Trp544 required for Mec1 activation are indicated.

(A) Kinase activity of Mec1 was detected using Rad53-kd as a substrate in buffer containing the indicated levels of NaCl. Increasing levels of Ddc1 peptide were added and phosphorylated Rad53-kd analyzed after 10 min.

(B) Ddc1 peptides with W352A or W544A mutations are inactive for Mec1 stimulation. The assay as in (A) was carried out in the presence of 60 mM NaCl.

(C) Quantification of Mec1 activation by Ddc1 peptides in (B), and those with W352Y, W352H, W352A, and W352R mutations. Average of three experiments with error bars are shown.