Martin G. Marinus - BMP Faculty - UMass Medical School

Recombinational Repair of Drug-Induced Damage. Cells have repair mechanisms that protect their DNA from the action of various cytotoxic agents. The agents we have studied are cisplatin, methylating agents and bleomycin, which are antitumor agents, and nitric oxide, which is thought to produce free radical damage to DNA. Although these compounds produce different types of DNA damage, we have found that homologous recombination is required to efficiently repair damage caused by these agents.

DNA Methylation.  Dam methyltransferase forms N⁶-methyladenine in GATC sequences in Escherichia coli. Such methylation is important for regulation of gene expression, initiation of chromosome replication and directionality of mismatch repair.
     In enterohemorrhagic E. coli, dam mutants show dramatic increases in adherence and and actin pedestal formation to cultured human cells compared to wildtype. This increase is accompanied by enhanced levels of certain virulence factors although the mRNA transcript levels of these factors are the same as wildtype. We are currently exploring the mechanism of this post-transcriptional effect.
    We have used oligonucleotide microarrays to determine global transcription patterns in cells with varying levels of Dam and SeqA proteins. The SeqA protein binds to a subset of dam sites. The results indicate that DNA methylation status of the cell is an important factor in forming and/or maintaining chromosome structure.

DNA Mismatch Repair and Antirecombination. Cells use mismatch repair to correct replication errors and a subset of mismatch repair proteins for antirecombination which prevents recombination between closely related DNA sequences. MutS protein, which is a multimer, binds to DNA mismatches to initiate either mismatch repair or antirecombination. To determine if specific MutS multimeric forms are important for function, we are characterizing mutant derivatives of MutS that have monomeric or different multimeric forms for their ability to perform mismatch repair or antirecombination.

Mismatch Repair-Induced Cell Death. Dam mutant cells are more sensitive to cisplatin and methylating agents than wildtype cells. If mismatch repair capacity is inactivated, however, the cells become resistant to these drugs. We are investigating the mechanism by which mismatch repair sensitizes cells to the action of these drugs.

Resource

Figure

Microarray results show that global gene expression in a seqA mutant strain (left panel) is the same as that in a wildtype strain overproducing the Dam methyltransferase (center panel) but that neither is similar to that for the dam mutant (right panel).

Representative Publications

Marinus, M.G., Casadesus, J. (2009) Roles of DNA adenine methylation in host-pathogen interactions: mismatch repair, transcriptional regulation, and more. FEMS Microbiol Rev. 33, 488-503.

Marinus, M. G., and A. Løbner-Olesen. 13 February 2009, posting date. Chapter 4.4.5, DNA Methylation. In A. Böck, R. Curtiss III, J. B. Kaper, P. D. Karp, F. C. Neidhardt, T. Nyström, J. M. Slauch, C. L. Squires, and D. Ussery (ed.), EcoSal—Escherichia coli and Salmonella: cellular and molecular biology. http://www.ecosal.org. ASM Press, Washington, DC.

Løbner-Olesen, A., Slominska-Wojewodzka, M., Hansen, F.G. and Marinus, M.G. (2008) DnaC inactivation in Escherichia coli K-12 induces the SOS response and expression of nucleotide biosynthesis genes. PLoS ONE 3, e2984.

Murphy, K.C., Ritchie, J.M., Waldor, M.K., Løbner-Olesen, A. and M.G. Marinus (2008) Dam methyltransferase is required for stable lysogeny of the Shiga Toxin (Stx2)-encoding bacteriophage 933W of enterohemorrhagic Escherichia coli O157:H7. J. Bacteriol. 190, 438-441 .

Nowosielska, A. and Marinus, M.G. (2008) DNA mismatch repair-induced double-strand breaks. DNA Repair 7, 48-56.

Marinus, M.G. (2008) Methylation and other Modifications of Nucleic Acids and Proteins. In Schaechter, M (ed.), Encyclopedia of Microbiology, Third Edition. Elsevier BV.

Broadbent,S.E., Balbontin, R., Casadesus, J., Marinus, M.G. and van der Woude, M. (2007) YhdJ, a non-essential CcrM-like DNA methyltransferase of Escherichia coli and Salmonella enterica. J. Bacteriol. 89, 4325-4327.

Campellone, K.G., Roe, A.J., Løbner-Olesen, A., Murphy, K.C., Magoun, L., Brady, M.J., Donohue-Rolfe, A.J., Tzipori, S., Gally, D.L., Leong, J.M. and Marinus, M. G. (2007) Increased adherence and actin pedestal formation by dam-deficient enterohemorrhagic Escherichia coli O157:H7. Molec. Microbiol. 63, 1468-1481 .

Riber, L., Olsson, J.A., Jensen, R.B., Skovgaard, O., Dasgupta, S., Marinus, M.G., and Lobner-Olesen, A. (2006) Hda-mediated inactivation of the DnaA protein and dnaA gene autoregulation act in concert to ensure homeostatic maintenance of the Escherichia coli chromosome. Genes Dev. 20, 2121-2134.

Nowosielska A, Smith SA, Engelward BP, Marinus MG. (2006) Homologous recombination prevents methylation-induced toxicity in Escherichia coli. Nucleic Acids Res. 34, 2258-68.

Lopez de Saro FJ, Marinus MG, Modrich P, O'Donnell M. (2006) The beta sliding clamp binds to multiple sites within MutL and MutS. J Biol Chem. 281, 14340-9.

Calmann, M.A. and Marinus, M.G. (2005) Differential effects of cisplatin and MNNG on dna mutants of Escherichia coli. Mutation Research 578, 406-416.

Calmann, M.A., Nowosielska, A. and Marinus, M.G. (2005) The MutS C-terminus is essential for mismatch repair activity in vivo. J. Bacteriol. 2005 187: 6577-6579.

Calmann, M.A., Evans, J.E. and Marinus, M. G. (2005) MutS inhibits RecA-mediated strand transfer with methylated DNA substrates. Nucleic Acids Research 33, 3591-3597.

Robbins-Manke, J.L,. Zdraveski, Z.Z., Marinus, M.G., and Essigmann, J.M. (2005) Analysis of global gene expression and double-strand-break formation in DNA adenine methyltransferase- and mismatch repair-deficient Escherichia coli. J. Bacteriol. 187, 7027-7037.

Nowosielska, A. and Marinus, M.G. (2005) Cisplatin induces DNA double-strand break formation in Escherichia coli dam mutants. DNA Repair 4, 773-781.

Løbner-Olesen, A., Skovgaard, O. and Marinus, M.G. (2005) Dam methylation: coordinating cellular processes. Current Opinion in Microbiology 8, 154-160.

Calmann, M.A., Nowosielska, A. and Marinus, M.G. (2005) Separation of mutation avoidance and antirecombination functions in an Escherichia coli mutS mutant. Nucleic Acids Research 33: 1193-1200.

Marinus, M.G. (2005) Dr. Jekyll and Mr. Hyde: How the MutSLH Repair System Kills the Cell, pp. 413-429. In Higgins, N.P (ed.) The Bacterial Chromosome. ASM Press, Washington DC.

Calmann, M.A. and Marinus, M.G. (2004) MutS inhibits RecA-mediated strand exchange with platinated DNA substrates. Proc.Natl.Acad.Sci. USA 101, 14174-14179.

Kosa, J.L., Zdraveski, Z.Z., Currier, C., Marinus, M.G., Essigmann, J.M. (2004) RecN and RecG are required for Escherichia coli survival of bleomycin-induced damage. Mutation Research 554, 149-157.

Nowosielska, A., Calmann, M.A., Zdraveski, Z., Essigmann, J.M. and Marinus, M.G. (2004) Spontaneous and cisplatin-induced recombination in Escherichia coli. DNA Repair 3, 719-728.

Calmann, M.A. and Marinus, M.G. (2003) Regulated expression of the Escherichia coli dam gene. J. Bacteriology 185, 5012-5014.

Løbner-Olesen, A., Marinus, M.G. and Hansen, F.G. (2003) Role of SeqA and Dam in Escherichia coli gene expression- a global/microarray analysis. Proc. Natl. Acad. Sci. USA 100, 4672-4677.

Roberts, R.J. et al (including Marinus, M.G.) (2003) A nomenclature for restriction enzymes, DNA methyltransferases, homing endonucleases and their genes. Nucleic Acids Research 31, 1805-1812.

Spek, E.J., Vuong, L.N., Matsuguchi, T., Marinus, M.G. and Engelward, B.P. (2002) Nitric oxide-induced homologous recombination in Escherichia coli is promoted by DNA glycosylases. J. Bacteriol. 184, 3501-3507.

Wu, T.H., Loh, T. and Marinus M.G. (2002) The function of Asp70, Glu77 and Lys79 in the Escherichia coli MutH protein. Nucleic Acids Res. 30, 818-822.

Zdraveski, Z.Z., Mello, J.A., Farinelli, C.K., Essigmann, J.M. and Marinus, M.G. (2002). MutS preferentially recognizes cisplatin- over oxaliplatin-modified DNA. J. Biol. Chem. 277, 1255-1260.

Loh, T., Murphy, K.C. and Marinus, M.G. (2001) Mutational analysis of the MutH protein from Escherichia coli. J. Biol. Chem. 276: 12113-12119.

Spek, E.J., Wright, T.I., Stitt, M.S., Taghizadeh, N.R., Tannenbaum, S.R., Marinus, M.G. and Engelward, B.P. (2001). Recombinational repair is critical for the survival of Escherichia coli exposed to nitric oxide. J. Bacteriol. 183, 131-138

Marinus, M.G. (2000) Recombination is essential for viability of an Escherichia coli dam (DNA adenine methyltransferase) mutant. J. Bacteriol., 182, 463-468.

Zdraveski, Z.Z., Mello, J.A., Marinus, M.G. and Essigmann, J.M. (2000) Multiple pathways of recombination define cellular responses to cisplatin. Chemistry and Biology, 7, 39-50.

Horst, J-.P., Wu, T-.H. and Marinus, M.G. (1999) Escherichia coli mutator genes. Trends in Microbiology. 7, 29-36.

Wu, T-.H. and Marinus, M.G. (1999) Deletion mutation analysis of the mutS gene in Escherichia coli. J. Biol. Chem. 274, 5948-5952.

Rotation Projects

Bleomycin is an anticancer drug that produces breaks in cellular DNA as well as damage to bases. Recombination-deficient cell lines show increased sensitivity towards this agent indicating that homologous recombination is essential to repair DNA damage.

1. Survey the known recombination-deficient strains for sensitivity to bleomycin to asses the importance of RecBCD versus RecFOR pathways.

3. Determine the number of DNA double-strand breaks as a function of bleomycin dose.

Academic Background

B.Sc.(Hons.), 1965, PhD, Otago, New Zealand, 1968
American Cancer Society Faculty Research Award, 1976-1981
Visiting Professor, University of Sussex, UK 1980-1981
Erskine Fellow, University of Canterbury, NZ 1991
Visiting Professor, University of Goettingen, Germany 1996-1997
Visiting Professor, Centre de Génétique Moléculaire, CNRS, Gif-sur-Yvette 2006

Office: LRB 917 , Lab 970F
Phone: 508-856-3330
E-mail: Martin.Marinus@umassmed.edu
Keywords: DNA Damage and Repair, DNA Replication, Genetics, DNA Recombination

Martin Marinus, Ph.D.

Research Home Page

DNA Repair and Recombination

Resource

Figure

Representative Publications

Rotation Projects

Academic Background