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. Author manuscript; available in PMC: 2013 Jul 31.
Published in final edited form as: Biochemistry. 2012 Jul 23;51(30):6028–6038. doi: 10.1021/bi3006412

Figure 2.

Figure 2

Reaction of UNG with free and histone-bound 147 bp 601 DNA containing global T→U substitutions. (a) Denaturing polyacrylamide gel separation of the DNA fragments resulting from base-catalyzed cleavage of the abasic sites generated from UNG excision of uracils. The DNA strand contained a 5′Fam-label and was reacted with hUNG for 0 s, 5 s, 10 s, 20 s, 30 s, 1 min, 2.5 min, 5 min, 10 min, 15 min, 30 min, 60 min and 95 min (lanes 1–13, left to right). The asterisk (*) indicates the migration position of a 5′ Fam-labeled DNA internal standard that was added post-reaction to normalize for lane loading differences (see Methods). (b) Same analysis as (a) except the DNA is in the context of the NCP. Reaction times are 0 s, 10 s, 32 s, 1 min, 2 min, 5 min, 10.9 min, 20 min, 30 min, 45 min, 60 min, 90 min, 120 min and 190 min (lanes 1–14, left to right). (c), (d) Relative activity of UNG for uracil sites in strand one and two of free and histone bound 601 DNA. The relative activity is defined as the initial rate of a uracil site in the NCP divided by that of the same site measured in the free DNA.