Gaasbeek, Esther J.;
van der Wal, Fimme J.;
van Putten, Jos P. M.;
de Boer, Paulo;
van der Graaf-van Bloois, Linda;
de Boer, Albert G.;
Vermaning, Bart J.;
Wagenaar, Jaap A.
Functional Characterization of Excision Repair and RecA-Dependent Recombinational DNA Repair in Campylobacter jejuni
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Media type:
E-Article
Title:
Functional Characterization of Excision Repair and RecA-Dependent Recombinational DNA Repair in Campylobacter jejuni
Contributor:
Gaasbeek, Esther J.;
van der Wal, Fimme J.;
van Putten, Jos P. M.;
de Boer, Paulo;
van der Graaf-van Bloois, Linda;
de Boer, Albert G.;
Vermaning, Bart J.;
Wagenaar, Jaap A.
Description:
<jats:title>ABSTRACT</jats:title>
<jats:p>
The presence and functionality of DNA repair mechanisms in
<jats:italic>Campylobacter jejuni</jats:italic>
are largely unknown. In silico analysis of the complete translated genome of
<jats:italic>C. jejuni</jats:italic>
NCTC 11168 suggests the presence of genes involved in methyl-directed mismatch repair (MMR), nucleotide excision repair, base excision repair (BER), and recombinational repair. To assess the functionality of these putative repair mechanisms in
<jats:italic>C. jejuni</jats:italic>
,
<jats:italic>mutS</jats:italic>
,
<jats:italic>uvrB</jats:italic>
,
<jats:italic>ung</jats:italic>
, and
<jats:italic>recA</jats:italic>
knockout mutants were constructed and analyzed for their ability to repair spontaneous point mutations, UV irradiation-induced DNA damage, and nicked DNA. Inactivation of the different putative DNA repair genes did not alter the spontaneous mutation frequency. Disruption of the UvrB and RecA orthologues, but not the putative MutS or Ung proteins, resulted in a significant reduction in viability after exposure to UV irradiation. Assays performed with uracil-containing plasmid DNA showed that the putative uracil-DNA glycosylase (Ung) protein, important for initiation of the BER pathway, is also functional in
<jats:italic>C. jejuni</jats:italic>
. Inactivation of
<jats:italic>recA</jats:italic>
also resulted in a loss of natural transformation. Overall, the data indicate that
<jats:italic>C. jejuni</jats:italic>
has multiple functional DNA repair systems that may protect against DNA damage and limit the generation of genetic diversity. On the other hand, the apparent absence of a functional MMR pathway may enhance the frequency of on-and-off switching of phase variable genes typical for
<jats:italic>C. jejuni</jats:italic>
and may contribute to the genetic heterogeneity of the
<jats:italic>C. jejuni</jats:italic>
population.
</jats:p>