Abstract
The main strategy used by pro-and eukaryotic cells for replication of damaged DNA is translesion synthesis (TLS). Here, we investigate the TLS process catalyzed by DNA polymerases β and λ on DNA substrates using mono-or dinucleotide gaps opposite damage located in the template strand. An analog of a natural apurinic/apyrimidinic site, the 3-hydroxy-2-hydroxymetylthetrahydrofuran residue (THF), was used as damage. DNA was synthesized in the presence of either Mg2+ or Mn2+. DNA polymerases β and λ were able to catalyze DNA synthesis across THF only in the presence of Mn2+. Moreover, strand displacement synthesis was not observed. The primer was elongated by only one nucleotide. Another unusual aspect of the synthesis is that dTTP could not serve as a substrate in all cases. dATP was a preferential substrate for synthesis catalyzed by DNA polymerase β. As for DNA polymerase λ, dGMP was the only incorporated nucleotide out of four investigated. Modified on heterocyclic base photoreactive analogs of dCTP and dUTP showed substrate specificity for DNA polymerase β. In contrast, the dCTP analog modified on the exocyclic amino group was a substrate for DNA polymerase λ. We also observed that human replication protein A inhibited polymerase incorporation by both DNA polymerases β and λ on DNA templates containing damage.
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Abbreviations
- AP-site:
-
apurinic/apyrimidinic site
- dRP:
-
deoxyribose phosphate
- hRPA:
-
human replication protein A
- THF:
-
3-hydroxy-2-hydroxymethyltetrahydrofuran
- TLS:
-
translesion synthesis
References
Goodman, M. F. (2002) Annu. Rev. Biochem., 71, 17–50.
Efrati, E., Tocco, G., Eritja, R., Wilson, S. H., and Goodman, M. F. (1997) J. Biol. Chem., 272, 2559–2569.
Ramadan, K., Shevelev, I. V., Maga, G., and Hubscher, U. (2002) J. Biol. Chem., 277, 18454–18458.
Wang, T. S. (1991) Annu. Rev. Biochem., 60, 513–552.
Sobol, R. W., Horton, J. K., Kuhn, R., Gu, H., Singhal, R. K., Prasad, R., Rajewsky, K., and Wilson, S. H. (1996) Nature, 379, 183–186.
Dianov, G. L., Prasad, R., Wilson, S. H., and Bohr, V. A. (1999) J. Biol. Chem., 274, 13741–13743.
Plug, A. W., Clairmont, C. A., Sapi, E., Ashley, T., and Sweasy, J. B. (1997) Proc. Natl. Acad. Sci. USA, 94, 1327–1331.
Sugo, N., Aratani, Y., Nagashima, Y., Kubota, Y., and Koyama, H. (2000) EMBO J., 19, 1397–1404.
Kunkel, T. A., and Alexander, P. S. (1986) J. Biol. Chem., 261, 160–166.
Singhal, R. K., and Wilson, S. H. (1993) J. Biol. Chem., 268, 15906–15911.
Ahn, J., Kraynov, V. S., Zhong, X. J., Werneburg, B. G., and Tsai, M. D. (1998) Biochem. J., 331, 79–87.
Vaisman, A., Warren, M. W., and Chaney, S. G. (2001) J. Biol. Chem., 276, 18999–19005.
Uchiyama, Y., Kimura, S., Yamamoto, T., Ishibashi, T., and Sakaguchi, K. (2004) Eur. J. Biochem., 271, 2799–2807.
Lebedeva, N. A., Rechkunova, N. I., Dezhurov, S. V., Khodyreva, S. N., Favre, A., Blanco, L., and Lavrik, O. I. (2005) Biochim. Biophys. Acta, 1751, 150–158.
Garcia-Diaz, M., Dominguez, O., Lopez-Fernandez, L. A., de Lera, L. T., Saniger, M. L., Ruiz, J. F., Parraga, M., Garcia-Ortiz, M. J., Kirchhoff, T., del Mazo, J., Bernard, A., and Blanco, L. (2000) J. Mol. Biol., 301, 851–867.
Lee, J. W., Blanco, L., Zhou, T., Garcia-Diaz, M., Bebenek, K., Kunkel, T. A., Wang, Z., and Povirk, L. F. (2004) J. Biol. Chem., 279, 805–811.
Garcia-Diaz, M., Bebenek, K., Sabariegos, R., Dominguez, O., Rodrigues, J., Kirchhoff, T., Garcia-Palomero, E., Picher, A. J., Juarez, R., Ruiz, J. F., Kunkel, T. A., and Blanco, L. (2002) J. Biol. Chem., 277, 13184–13191.
Lindahl, T., and Wood, R. D. (1999) Science, 286, 1897–1905.
Maga, G., Villani, G., Crespan, E., Wimmer, U., Ferrari, E., Bertocci, B., and Hubscher, U. (2007) Nature, 447, 606–608.
Cox, M. M. (2001) Annu. Rev. Biochem., 71, 71–100.
Wilson, D. M., III (2005) J. Mol. Biol., 345, 1003–1014.
Cahill, D., Connor, B., and Carney, J. P. (2006) Front. Biosci., 11, 1958–1976.
Braithwaite, E. K., Prasad, R., Shock, D. D., Hou, E. W., Beard, W. A., and Wilson, S. H. (2005) J. Biol. Chem., 280, 18469–18475.
Date, T., Yamaguchi, M., Hirose, F., Nishimoto, Y., Tanihara, K., and Matsukage, A. (1998) Biochemistry, 27, 2983–2990.
Wlassoff, W. A., Dobrikov, M. I., Safronov, I. V., Dudko, R. Y., Bogachev, V. S., Kandaurova, V. V., Shishkin, G. V., Dymshits, G. M., and Lavrik, O. I. (1995) Bioconjug. Chem., 6, 352–360.
Kolpashchikov, D. M., Zakharenko, A. L., Dezhurov, S. V., Rechkunova, N. I., Khodyreva, S. N., Degtyarev, S. Kh., Litvak, V. V., and Lavrik, O. I. (1999) Bioorg. Khim., 25, 129–136.
Dezhurov, S. V., Khodyreva, S. N., Plekhanova, E. S., and Lavrik, O. I. (2005) Bioconjug. Chem., 16, 215–222.
Yamshchikov, V. F. (1990) in Methods of Molecular Genetics and Gene Engineering (Salganik, R. I., ed.) [in Russian], Nauka, Novosibirsk, p. 28.
Yamshchikov, V. F. (1990) in Methods of Molecular Genetics and Gene Engineering (Salganik, R. I., ed.) [in Russian], Nauka, Novosibirsk, pp. 145–154.
Khodyreva, S. N., and Lavrik, O. I. (2005) Curr. Med. Chem., 12, 641–655.
Blanca, G., Villani, G., Shevelev, I., Ramadan, K., Spadari, S., Hubscher, U., and Maga, G. (2004) Biochemistry, 43, 11605–11615.
Taylor, J. S. (2002) Mutat. Res., 510, 55–70.
Kunkel, T. A., and Alexander, S. J. (1986) J. Biol. Chem., 261, 160–166.
Maga, G., Villani, G., Ramadan, K., Shevelev, I., le Gac, N. T., Blanco, L., Blanca, G., Spadari, S., and Hubscher, U. (2002) J. Biol. Chem., 277, 48434–48440.
Maga, G., Shevelev, I., Villani, Spadari, S., and Hubscher, U. (2006) Nucleic Acids Res., 34, 1405–1415.
Crespan, E., Hubscher, U., and Maga, G. (2007) Nucleic Acids Res., 35, 5173–1581.
Krasikova, Yu. S., Belousova, E. A., Lebedeva, N. A., Pestryakov, P. E., and Lavrik, O. I. (2008) Biochemistry (Moscow), 73, 1042–1046.
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Original Russian Text © A. A. Shtygasheva, E. A. Belousova, N. I. Rechkunova, N. A. Lebedeva, O. I. Lavrik, 2008, published in Biokhimiya, 2008, Vol. 73, No. 11, pp. 1504–1512.
Originally published in Biochemistry (Moscow) On-Line Papers in Press, as Manuscript BM07-440, October 19, 2008.
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Shtygasheva, A.A., Belousova, E.A., Rechkunova, N.I. et al. DNA polymerases β and λ as potential participants of TLS during genomic DNA replication on the lagging strand. Biochemistry Moscow 73, 1207–1213 (2008). https://doi.org/10.1134/S0006297908110060
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DOI: https://doi.org/10.1134/S0006297908110060