title: Mismatch repair in Trypanosoma brucei: heterologous expression of MSH2 from Trypanosoma cruzi provides new insights into the response to oxidative damage.
abstract: Trypanosomes are unicellular eukaryotes that cause disease in humans and other mammals. Trypanosoma cruzi and Trypanosoma brucei are the causative agents, respectively, of Chagas disease in the Americas and sleeping sickness in sub-Saharan Africa. To better comprehend the interaction of these parasites with their hosts, understanding the mechanisms involved in the generation of genetic variability is critical. One such mechanism is mismatch repair (MMR), which has a crucial, evolutionarily conserved role in maintaining the fidelity of DNA replication, as well as acting in other cellular processes, such as DNA recombination. Here we have attempted to complement T. brucei MMR through the expression of MSH2 from T. cruzi. Our results show that T. brucei MSH2-null mutants are more sensitive to hydrogen peroxide (H2O2) than wild type cells, suggesting the involvement of MSH2 in the response to oxidative stress in this parasite. This phenotype is reverted by the expression of either the T. cruzi or the T. brucei MSH2 protein in the MSH2-null mutants. In contrast, MMR complementation, as assessed by resistance to N-methyl-N'-nitro-N-nitrosoguanidine (MNNG) and microsatellite instability, was not achieved by the heterologous expression of T. cruzi MSH2. This finding, associated to the demonstration that mutation of MLH1, another component of the MMR system, did not affect sensitivity of T. brucei cells to H2O2, suggests an additional role of MSH2 in dealing with oxidative damage in these parasites, which may occur independently of MMR.
authors.affiliation: Instituto de Ciências Biológicas, Universidade Federal de Minas Gerais, Departamento de Bioquímica e Imunologia, Belo Horizonte, Brazil.
authors.forename: Alice
authors.initials: A
authors.lastname: Machado-Silva
authors.forename: Santuza M R
authors.initials: SM
authors.lastname: Teixeira
authors.forename: Glória R
authors.initials: GR
authors.lastname: Franco
authors.forename: Andréa M
authors.initials: AM
authors.lastname: Macedo
authors.forename: Sérgio D J
authors.initials: SD
authors.lastname: Pena
authors.forename: Richard
authors.initials: R
authors.lastname: McCulloch
authors.forename: Carlos Renato
authors.initials: CR
authors.lastname: Machado
mesh_terms: D000818:Animals; D004249:DNA Damage; D053843:DNA Mismatch Repair; D017125:Genes, Protozoan; D006861:Hydrogen Peroxide / Q000378:metabolism / Q000494:pharmacology; D053842:Microsatellite Instability; D051718:MutS Homolog 2 Protein / Q000235:genetics*; D009154:Mutation; D018384:Oxidative Stress; D015800:Protozoan Proteins / Q000235:genetics*; D011995:Recombination, Genetic; D014346:Trypanosoma brucei brucei / Q000235:genetics*; D014349:Trypanosoma cruzi / Q000235:genetics*
chemical_list: D015800:Protozoan Proteins; D006861:Hydrogen Peroxide; C435509:MSH2 protein, Trypanosoma cruzi; D051718:MutS Homolog 2 Protein