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. 2017 Feb 1;45(6):3378–3394. doi: 10.1093/nar/gkx066

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© The Author(s) 2017. Published by Oxford University Press on behalf of Nucleic Acids Research.

This is an Open Access article distributed under the terms of the Creative Commons Attribution License (http://creativecommons.org/licenses/by-nc/4.0/), which permits non-commercial re-use, distribution, and reproduction in any medium, provided the original work is properly cited. For commercial re-use, please contact journals.permissions@oup.com

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Figure 1. — Analysis of A3C processivity on ssDNA oligonucleotides. Processivity of A3C was tested on ssDNA substrates that contain a fluorescein-labeled deoxythymidine (yellow star) between two 5΄TTC deamination motifs separated by different distances. (A and B) hA3C S188I is more processive than hA3C. (A) Deamination of a 60 nt ssDNA substrate with deamination targets spaced 5 nt apart. Single deaminations of the 5΄C and 3΄C are detected as the appearance of labeled 42- and 23-nt fragments, respectively; double deamination of both C residues on the same molecule results in a 5 nt labeled fragment. (B) Deamination of a 118 nt ssDNA substrate with deaminated cytosines spaced 63 nt apart. Single deaminations of the 5΄C and 3΄C are detected as the appearance of labeled 100- and 81-nt fragments, respectively; double deamination of both C residues on the same molecule results in a 63 nt labeled fragment. (C–F) cA3C and gA3C are more processive than hA3C. (C) Deamination of a 60 nt ssDNA substrate as for panel (A). (D) Deamination of a 118 nt ssDNA as for panel (B). (E) Deamination of a 69 nt ssDNA substrate with deamination targets spaced 14 nt apart. Single deaminations of the 5΄C and 3΄C are detected as the appearance of labeled 51- and 32-nt fragments, respectively; double deamination of both C residues on the same molecule results in a 14 nt labeled fragment. (F) Deamination of an 85 nt ssDNA substrate with deaminated cytosines spaced 30 nt apart. Single deaminations of the 5΄C and 3΄C are detected as the appearance of labeled 67- and 48-nt fragments, respectively; double deamination of both C residues on the same molecule results in a 30 nt labeled fragment. If no 5΄C and 3΄C band was detected, the processivity was denoted with N.D. (not detected). The measurements of enzyme processivity (processivity factor) and the S.D. are shown below the gels. All values are calculated from at least three independent experiments.