Calcium inhibits bap-dependent multicellular behavior in Staphylococcus aureus

Bap (biofilm-associated protein) is a 254-kDa staphylococcal surface protein implicated in formation of biofilms by staphylococci isolated from chronic mastitis infections. The presence of potential EF-hand motifs in the amino acid sequence of Bap prompted us to investigate the effect of calcium on...

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Autores principales: Arrizubieta, M. J., Toledo-Arana, Alejandro, Amorena, B., Penadés, José R., Lasa, Inigo
Formato: article
Lenguaje:Inglés
Publicado: 2017
Acceso en línea:http://hdl.handle.net/20.500.11939/4450
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author Arrizubieta, M. J.
Toledo-Arana, Alejandro
Amorena, B.
Penadés, José R.
Lasa, Inigo
author_browse Amorena, B.
Arrizubieta, M. J.
Lasa, Inigo
Penadés, José R.
Toledo-Arana, Alejandro
author_facet Arrizubieta, M. J.
Toledo-Arana, Alejandro
Amorena, B.
Penadés, José R.
Lasa, Inigo
author_sort Arrizubieta, M. J.
collection ReDivia
description Bap (biofilm-associated protein) is a 254-kDa staphylococcal surface protein implicated in formation of biofilms by staphylococci isolated from chronic mastitis infections. The presence of potential EF-hand motifs in the amino acid sequence of Bap prompted us to investigate the effect of calcium on the multicellular behavior of Bap-expressing staphylococci. We found that addition of millimolar amounts of calcium to the growth media inhibited intercellular adhesion of and biofilm formation by Bap-positive strain V329. Addition of manganese, but not addition of magnesium, also inhibited biofilm formation, whereas bacterial aggregation in liquid media was greatly enhanced by metal-chelating agents. In contrast, calcium or chelating agents had virtually no effect on the aggregation of Bap-deficient strain M556. The biofilm elicited by insertion of bap into the chromosome of a biofilm-negative strain exhibited a similar dependence on the calcium concentration, indicating that the observed calcium inhibition was an inherent property of the Bap-mediated biofilms. Site-directed mutagenesis of two of the putative EF-hand domains resulted in a mutant strain that was capable of forming a biofilm but whose biofilm was not inhibited by calcium. Our results indicate that Bap binds Ca2+ with low affinity and that Ca2+ binding renders the protein noncompetent for biofilm formation and for intercellular adhesion. The fact that calcium inhibition of Bap-mediated multicellular behavior takes place in vitro at concentrations similar to those found in milk serum supports the possibility that this inhibition is relevant to the pathogenesis and/or epidemiology of the bacteria in the mastitis process.
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spelling ReDivia44502025-04-25T14:43:34Z Calcium inhibits bap-dependent multicellular behavior in Staphylococcus aureus Arrizubieta, M. J. Toledo-Arana, Alejandro Amorena, B. Penadés, José R. Lasa, Inigo Bap (biofilm-associated protein) is a 254-kDa staphylococcal surface protein implicated in formation of biofilms by staphylococci isolated from chronic mastitis infections. The presence of potential EF-hand motifs in the amino acid sequence of Bap prompted us to investigate the effect of calcium on the multicellular behavior of Bap-expressing staphylococci. We found that addition of millimolar amounts of calcium to the growth media inhibited intercellular adhesion of and biofilm formation by Bap-positive strain V329. Addition of manganese, but not addition of magnesium, also inhibited biofilm formation, whereas bacterial aggregation in liquid media was greatly enhanced by metal-chelating agents. In contrast, calcium or chelating agents had virtually no effect on the aggregation of Bap-deficient strain M556. The biofilm elicited by insertion of bap into the chromosome of a biofilm-negative strain exhibited a similar dependence on the calcium concentration, indicating that the observed calcium inhibition was an inherent property of the Bap-mediated biofilms. Site-directed mutagenesis of two of the putative EF-hand domains resulted in a mutant strain that was capable of forming a biofilm but whose biofilm was not inhibited by calcium. Our results indicate that Bap binds Ca2+ with low affinity and that Ca2+ binding renders the protein noncompetent for biofilm formation and for intercellular adhesion. The fact that calcium inhibition of Bap-mediated multicellular behavior takes place in vitro at concentrations similar to those found in milk serum supports the possibility that this inhibition is relevant to the pathogenesis and/or epidemiology of the bacteria in the mastitis process. 2017-06-01T10:10:10Z 2017-06-01T10:10:10Z 2004 NOV 2004 article Arrizubieta, M.J., Toledo-Arana, A., Amorena, B., Penades, J.R., Lasa, I. (2004). Calcium inhibits bap-dependent multicellular behavior in Staphylococcus aureus. Journal of Bacteriology, 186(22), 7490-7498. 0021-9193 http://hdl.handle.net/20.500.11939/4450 10.1128/JB.186.22.7490-7498.2004 en openAccess Impreso
spellingShingle Arrizubieta, M. J.
Toledo-Arana, Alejandro
Amorena, B.
Penadés, José R.
Lasa, Inigo
Calcium inhibits bap-dependent multicellular behavior in Staphylococcus aureus
title Calcium inhibits bap-dependent multicellular behavior in Staphylococcus aureus
title_full Calcium inhibits bap-dependent multicellular behavior in Staphylococcus aureus
title_fullStr Calcium inhibits bap-dependent multicellular behavior in Staphylococcus aureus
title_full_unstemmed Calcium inhibits bap-dependent multicellular behavior in Staphylococcus aureus
title_short Calcium inhibits bap-dependent multicellular behavior in Staphylococcus aureus
title_sort calcium inhibits bap dependent multicellular behavior in staphylococcus aureus
url http://hdl.handle.net/20.500.11939/4450
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