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Infection and Immunity, August 2001, p. 5098-5106, Vol. 69, No. 8
0019-9567/01/$04.00+0 DOI: 10.1128/IAI.69.8.5098-5106.2001
Copyright © 2001, American Society for Microbiology. All rights reserved.
Contribution of Mn-Cofactored Superoxide Dismutase
(SodA) to the Virulence of Streptococcus
agalactiae
Claire
Poyart,1,2,*
Elisabeth
Pellegrini,1
Olivier
Gaillot,1
Claire
Boumaila,1
Marina
Baptista,1,2 and
Patrick
Trieu-Cuot1,2
INSERM U-4111 and
Laboratoire Mixte Pasteur-Necker de Recherche sur les
Streptocoques et Streptococcies,2
Faculté de Médecine Necker-Enfants Malades, 75730 Paris
Cedex 15, France
Received 27 November 2000/Returned for modification 26 February
2001/Accepted 14 May 2001
Superoxide dismutases convert superoxide anions to molecular oxygen
and hydrogen peroxide, which, in turn, is metabolized by catalases
and/or peroxidases. These enzymes constitute one of the major defense
mechanisms of cells against oxidative stress and hence play a role in
the pathogenesis of certain bacteria. We previously demonstrated that
group B streptococci (GBS) possess a single Mn-cofactored superoxide
dismutase (SodA). To analyze the role of this enzyme in the
pathogenicity of GBS, we constructed a sodA-disrupted
mutant of Streptococcus agalactiae NEM316 by allelic
exchange. This mutant was subsequently cis complemented by
integration into the chromosome of pAT113/Sp harboring the wild-type
sodA gene. The SOD specific activity detected by gel analysis in cell extracts confirmed that active SODs were present in
the parental and complemented strains but absent in the
sodA mutant. The growth rates of these strains in standing
cultures were comparable, but the sodA mutant was extremely
susceptible to the oxidative stress generated by addition of paraquat
or hydrogen peroxide to the culture medium and exhibited a higher
mutation frequency in the presence of rifampin. In mouse bone
marrow-derived macrophages, the sodA mutant showed an
increased susceptibility to bacterial killing by macrophages. In a
mouse infection model, after intravenous injection the survival of the
sodA mutant in the blood and the brain was markedly reduced
in comparison to that of the parental and complemented strains whereas
only minor effects on survival in the liver and the spleen were
observed. These results suggest that SodA plays a role in GBS pathogenesis.
*
Corresponding author. Mailing address: Laboratoire de
Microbiologie, Faculté de Médecine Necker-Enfants Malades,
156 rue de Vaugirard, 75730 Paris Cedex 15, France. Phone: (33) (1) 40 61 56 79. Fax: (33) (1) 40 61 55 92. E-mail:
cpoyart{at}pasteur.fr.
Infection and Immunity, August 2001, p. 5098-5106, Vol. 69, No. 8
0019-9567/01/$04.00+0 DOI: 10.1128/IAI.69.8.5098-5106.2001
Copyright © 2001, American Society for Microbiology. All rights reserved.
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