TY - JOUR
T1 - Inhibition of DNA replication by an anti-PCNA aptamer/PCNA complex
AU - Kowalska, Ewa
AU - Bartnicki, Filip
AU - Fujisawa, Ryo
AU - Bonarek, Piotr
AU - Hermanowicz, Pawet
AU - Tsurimoto, Toshiki
AU - Muszynska, Klaudia
AU - Strzalka, Wojciech
N1 - Funding Information:
We thank Agnieszka Banas, ´ Justyna Labuz - and Piotr Zg-lobicki for comments on this manuscript. National Centre of Research and Development [LIDER/28/54/L-2/10/NCBiR/2011 to W.S.]; Ministry of Science and Higher Education. Funding for open access charge: The Faculty of Biochemistry, Biophysics and Biotechnology of Jagiellonian University is a partner of the Leading National Research Center (KNOW) supported by the Ministry of Science and Higher Education.
Funding Information:
National Centre of Research and Development [LIDER/28/54/L-2/10/NCBiR/2011 to W.S.]; Ministry of Science and Higher Education. Funding for open access charge: The Faculty of Biochemistry, Biophysics and Biotechnology of Jagiellonian University is a partner of the Leading National Research Center (KNOW) supported by the Ministry of Science and Higher Education. Conflict of interest statement. None declared.
Publisher Copyright:
C The Author(s) 2017. Published by Oxford University Press on behalf of Nucleic Acids Research.
PY - 2018/1/9
Y1 - 2018/1/9
N2 - Proliferating cell nuclear antigen (PCNA) is a multifunctional protein present in the nuclei of eukaryotic cells that plays an important role as a component of the DNA replication machinery, as well as DNA repair systems. PCNA was recently proposed as a potential non-oncogenic target for anti-cancer therapy. In this study, using the Systematic Evolution of Ligands by EXponential enrichment (SELEX) method, we developed a short DNA aptamer that binds human PCNA. In the presence of PCNA, the anti-PCNA aptamer inhibited the activity of human DNA polymerase and at nM concentrations. Moreover, PCNA protected the anti-PCNA aptamer against the exonucleolytic activity of these DNA polymerases. Investigation of the mechanism of anti-PCNA aptamer-dependent inhibition of DNA replication revealed that the aptamer did not block formation, but was a component of PCNA/DNA polymerase or complexes. Additionally, the anti-PCNA aptamer competed with the primer-template DNA for binding to the PCNA/DNA polymerase or complex. Based on the observations, a model of anti-PCNA aptamer/PCNA complex-dependent inhibition of DNA replication was proposed.
AB - Proliferating cell nuclear antigen (PCNA) is a multifunctional protein present in the nuclei of eukaryotic cells that plays an important role as a component of the DNA replication machinery, as well as DNA repair systems. PCNA was recently proposed as a potential non-oncogenic target for anti-cancer therapy. In this study, using the Systematic Evolution of Ligands by EXponential enrichment (SELEX) method, we developed a short DNA aptamer that binds human PCNA. In the presence of PCNA, the anti-PCNA aptamer inhibited the activity of human DNA polymerase and at nM concentrations. Moreover, PCNA protected the anti-PCNA aptamer against the exonucleolytic activity of these DNA polymerases. Investigation of the mechanism of anti-PCNA aptamer-dependent inhibition of DNA replication revealed that the aptamer did not block formation, but was a component of PCNA/DNA polymerase or complexes. Additionally, the anti-PCNA aptamer competed with the primer-template DNA for binding to the PCNA/DNA polymerase or complex. Based on the observations, a model of anti-PCNA aptamer/PCNA complex-dependent inhibition of DNA replication was proposed.
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U2 - 10.1093/nar/gkx1184
DO - 10.1093/nar/gkx1184
M3 - Article
C2 - 29186524
AN - SCOPUS:85045958275
SN - 0305-1048
VL - 46
SP - 25
EP - 41
JO - Nucleic acids research
JF - Nucleic acids research
IS - 1
ER -
