RESUMO
The Escherichia coli SeqA protein, a negative regulator of chromosome DNA replication, prevents the overinitiation of replication within one cell cycle by binding to hemimethylated GATC sequences in the replication origin, oriC. In addition to the hemimethylated DNA-binding activity, the SeqA protein has a self-association activity, which is also considered to be essential for its regulatory function in replication initiation. To study the SeqA protein biological activity, we performed a SeqA protein model to examine its architecture. SeqA has a bipartite structure composed of a large and small lobe. The SeqA spatial conformation contributes to its ability to bind to a pair of hemimethylated GATC sequences and to its cooperative behavior.
Assuntos
Proteínas da Membrana Bacteriana Externa/química , Proteínas da Membrana Bacteriana Externa/metabolismo , DNA Bacteriano/química , Proteínas de Ligação a DNA/química , Proteínas de Ligação a DNA/metabolismo , Proteínas de Escherichia coli/química , Proteínas de Escherichia coli/metabolismo , Replicação do DNA , Dimerização , Estabilidade Enzimática , Escherichia coli/genética , Escherichia coli/metabolismo , Modelos Moleculares , Conformação Proteica , Dobramento de ProteínaRESUMO
Numerous studies have shown that Salmonella typhimurium dam mutants are highly attenuated for virulence in mice. In vivo studies have also shown that, on oral and intraperitoneal administration, low number of these mutants is able to colonize and persist in target organs. So, they must sense and overcome a myriad of host killing mechanisms. Our goal was to evaluate the effect of in vivo passage, in mice, on S. typhimurium dam mutant virulence and fitness. Swiss albino mice were used for the determination of LD50 and enumeration of bacteria recovered from liver eight days postinfection. Our results indicate that LD50 values of re-isolated mutants were at least two to three-fold lower than those of control strains. Strains re-isolated from liver showed decreased in vitro sensitivity toward sodium deoxycholate and H(2)O(2) than control strains. In addition, the number of re-isolated mutants colonizing spleen and liver was relatively higher than control strains. According to these results, we suggest that persistence of S. typhimurium dam mutants within target organs can increase their in vivo virulence in mice.