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. Author manuscript; available in PMC: 2012 Sep 2.
Published in final edited form as: Cell. 2012 Feb 23;148(5):922–932. doi: 10.1016/j.cell.2012.01.033

Figure 3.

Figure 3

Ku80Δ28 Protein Loses Affinity for Both dsDNA and TLC1-KBS RNA

(A) Binding of 22-bp dsDNA with a 14-nt telomeric 3' overhang by purified WT Ku and Ku80Δ28 proteins assayed by EMSA. (Throughout this paper, Ku80Δ28 protein refers to a heterodimer of Ku70 and Ku80Δ28.) Lanes 1 – 6 contain the following amounts of active WT Ku protein: 0; 0.02 nM; 0.2 nM; 1.7 nM; 17.2 nM; 172 nM. Lanes 7 – 12 contain active Ku80Δ28 protein in the same amounts as in lanes 1 – 6.

(B) Binding of TLC1-KBS RNA to WT Ku and Ku80Δ28 assayed via EMSA. The amounts of active protein are the same as in (A).

(C) Graphical representation of the DNA binding seen in (A). The active Kd for WT Ku is 0.08 nM. The active Kd for Ku80Δ28 is 0.78 nM.

(D) Graphical representation of (B). The fitted data yielded active Kd = 4.9 nM for WT Ku and active Kd = 87.1 nM for Ku80Δ28 binding to TLC1-KBS RNA. See also Figure S3.