In vitro affinity of Deinococcus radiodurans MutS towards mismatched DNA exceeds that of its orthologues from Escherichia coli and Thermus thermophilus - Publication - MOST Wiedzy


In vitro affinity of Deinococcus radiodurans MutS towards mismatched DNA exceeds that of its orthologues from Escherichia coli and Thermus thermophilus


The mismatch binding protein MutS is responsible for the recognition of mispaired and unpaired bases, which is the initial step in DNA repair. Among the MutS proteins most extensively studied in vitro are those derived from Thermus thermophilus, Thermus aquaticus and Escherichia coli. Here, we present the first report on the in vitro examination of DNA mismatch binding activity of MutS protein from Deinococcus radiodurans and confront this with the properties of those from E. coli and T. thermophilus. The analyses which included mobility gel-shift assay, colorimetric and qPCR estimation of MutS-bound DNA clearly showed that D. radiodurans MutS exhibited much higher affinity towards mismatched DNA in vitro than its counterparts from E. coli and T. thermophilus. In addition, D. radiodurans MutS displayed a significantly higher specificity of DNA mismatch binding than the two other orthologues. The specificity expressed as the ratio of mismatched to fully complementary DNA bound reached over 4 and 20-fold higher values for D. radiodurans than for T. thermophilus and E. coli MutS, respectively. The results demonstrate mainly the biotechnological potential of D. radiodurans MutS but the in vitro characteristics of the MutS orthologues could reflect substantial differences in DNA mismatch binding activities existing in vivo.


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JOURNAL OF BIOTECHNOLOGY no. 252, pages 55 - 64,
ISSN: 0168-1656
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Bibliographic description:
Banasik M., Stanisławska-Sachadyn A., Hildebrandt E., Sachadyn P.: In vitro affinity of Deinococcus radiodurans MutS towards mismatched DNA exceeds that of its orthologues from Escherichia coli and Thermus thermophilus// JOURNAL OF BIOTECHNOLOGY. -Vol. 252, (2017), s.55-64
Digital Object Identifier (open in new tab) 10.1016/j.jbiotec.2017.05.010
Bibliography: test
  1. Banasik, M., Sachadyn, P., 2016. A colorimetric microplate assay for DNA-binding activity of his-tagged MutS protein. Mol. Biotechnol. 58, 521-527. open in new tab
  2. Battista, J.R., Earl, A.M., Park, M.J., 1999. Why is Deinococcus radiodurans so resistant to ionizing radiation? Trends Microbiol. 7, 362-365. open in new tab
  3. Binkowski, B.F., Richmond, K.E., Kaysen, J., Sussman, M.R., Belshaw, P.J., 2005. Correcting errors in synthetic DNA through consensus shuffling. Nucleic Acids Res. 33, e55. open in new tab
  4. Biswas, I., Hsieh, P., 1996. Identification and characterization of a thermostable MutS homolog from Thermus aquaticus. J. Biol. Chem. 271, 5040-5048.
  5. Blackwell, L.J., Bjornson, K.P., Allen, D.J., Modrich, P., 2001. Distinct MutS DNA-binding modes that are differentially modulated by ATP binding and hydrolysis. J. Biol. Chem. 276, 34339-34347. open in new tab
  6. Brown, J., Brown, T., Fox, K.R., 2001. Affinity of mismatch-binding protein MutS for heteroduplexes containing different mismatches. Biochem. J. 354, 627-633. open in new tab
  7. Cho, M., Chung, S., Heo, S.D., Ku, J., Ban, C., 2007. A simple fluorescent method for detecting mismatched DNAs using a MutS-fluorophore conjugate. Biosens. Bioelectron. 22, 1376-1381. open in new tab
  8. Cox, M.M., Battista, J.R., 2005. Deinococcus radiodurans-the consummate survivor. Nat. Rev. Microbiol. 3, 882-892. open in new tab
  9. Fukui, K., Bessho, Y., Shimada, A., Yokoyama, S., Kuramitsu, S., 2013. Thermostable mismatch-recognizing protein MutS suppresses nonspecific amplification during polymerase chain reaction (PCR). Int. J. Mol. Sci. 14, 6436-6453. open in new tab
  10. Ito, H., Watanabe, H., Takehisa, M., Iizuka, H., 1983. Isolation and identification of radiation-resistant cocci belonging to the genus Deinococcus from sewage sludges and animal feeds. Agric. Biol. Chem. 47, 1239-1247. open in new tab
  11. Jiricny, J., 2013. Postreplicative mismatch repair. Cold Spring Harb. Perspect. Biol. 5, a012633. open in new tab
  12. Joshi, A., Rao, B.J., 2001. MutS recognition: multiple mismatches and sequence context effects. J. Biosci. 26, 595-606. open in new tab
  13. Kunkel, T.A., Erie, D.A., 2005. DNA mismatch repair. Annu. Rev. Biochem. 74, 681-710. open in new tab
  14. Lamers, M.H., Perrakis, A., Enzlin, J.H., Winterwerp, H.H., de Wind, N., Sixma, T.K., 2000. The crystal structure of DNA mismatch repair protein MutS binding to a G x T mismatch. Nature 407, 711-717. open in new tab
  15. Lishanski, A., Ostrander, E.A., Rine, J., 1994. Mutation detection by mismatch binding protein, MutS, in amplified DNA: application to the cystic fibrosis gene. Proc. Natl. Acad. Sci. U. S. A. 91, 2674-2678. open in new tab
  16. Lu, A.L., Clark, S., Modrich, P., 1983. Methyl-directed repair of DNA base-pair mismatches in vitro. Proc. Natl. Acad. Sci. U. S. A. 80, 4639-4643. open in new tab
  17. Mennecier, S., Coste, G., Servant, P., Bailone, A., Sommer, S., 2004. Mismatch repair ensures fidelity of replication and recombination in the radioresistant organism Deinococcus radiodurans. Mol. Genet. Genom.: MGG 272, 460-469. open in new tab
  18. Minton, K.W., 1994. DNA repair in the extremely radioresistant bacterium Deinococcus radiodurans. Mol. Microbiol. 13, 9-15. open in new tab
  19. Moseley, B.E., Mattingly, A., 1971. Repair of irradiation transforming deoxyribonucleic acid in wild type and a radiation-sensitive mutant of Micrococcus radiodurans. J. Bacteriol. 105, 976-983. open in new tab
  20. Obmolova, G., Ban, C., Hsieh, P., Yang, W., 2000. Crystal structures of mismatch repair protein MutS and its complex with a substrate DNA. Nature 407, 703-710. open in new tab
  21. Olchowy, J., Kur, K., Sachadyn, P., Milewski, S., 2006. Construction, purification, and functional characterization of his-tagged Candida albicans glucosamine-6-phosphate synthase expressed in Escherichia coli. Protein Expr. Purif. 46, 309-315. open in new tab
  22. Sachadyn, P., 2010. Conservation and diversity of MutS proteins. Mutat. Res. 694, 20-30. open in new tab
  23. Sixma, T.K., 2001. DNA mismatch repair: MutS structures bound to mismatches. Curr. Opin. Struct. Biol. 11, 47-52. open in new tab
  24. Stanislawska-Sachadyn, A., Sachadyn, P., Jedrzejczak, R., Kur, J., 2003. Construction and purification of his6-Thermus thermophilus MutS protein. Protein Expr. Purif. 28, 69-77. open in new tab
  25. Stanislawska-Sachadyn, A., Paszko, Z., Kluska, A., Skasko, E., Sromek, M., Balabas, A., Janiec-Jankowska, A., Wisniewska, A., Kur, J., Sachadyn, P., 2005. Preliminary studies on DNA retardation by MutS applied to the detection of point mutations in clinical samples. Mutat. Res. 570, 97-103. open in new tab
  26. Stanislawska-Sachadyn, A., Sachadyn, P., Ihle, K., Sydorczuk, C., Wiejacha, K., Kur, J., 2006. The construction of bifunctional fusion proteins consisting of MutS and GFP. J. Biotechnol. 121, 134-143. open in new tab
  27. Takamatsu, S., Kato, R., Kuramitsu, S., 1996. Mismatch DNA recognition protein from an extremely thermophilic bacterium, Thermus thermophilus HB8. Nucleic Acids Res. 24, 640-647. open in new tab
  28. Whitehouse, A., Deeble, J., Parmar, R., Taylor, G.R., Markham, A.F., Meredith, D.M., 1997. Analysis of the mismatch and insertion/deletion binding properties of Thermus thermophilus, HB8, MutS. Biochem. Biophys. Res. Commun. 233, 834-837. open in new tab
  29. Zhong, T., Zhou, Y., Bi, L., Zhang, X.E., 2011. MutS-mediated enrichment of mutated DNA produced by directed evolution in vitro. World J. Microbiol. Biotechnol. 27, 1367-1372. open in new tab
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