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bioRxiv · 10.1101/2021.05.28.446192

Manganese transport by Streptococcus sanguinis in acidic conditions and its impact on growth in vitro and in vivo

Puccio, T.·Schultz, A. C.·Lizarraga, C. A.·Bryant, A. S.·Culp, D. J.·Burne, R.·Kitten, T.
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Abstract

Streptococcus sanguinis is an oral commensal and an etiological agent of infective endocarditis. Previous studies have identified the SsaACB manganese transporter as essential for endocarditis virulence; however, the significance of SsaACB in the oral environment has never been examined. Here we report that a {Delta}ssaACB mutant of strain SK36 exhibits reduced growth and manganese uptake under acidic conditions. Further studies revealed that these deficits resulted from the decreased activity of TmpA, shown in the accompanying paper to function as a ZIP-family manganese transporter. Transcriptomic analysis of fermentor-grown cultures of SK36 WT and {Delta}ssaACB strains identified pH-dependent changes related to carbon catabolite repression in both strains, though their magnitude was generally greater in the mutant. In strain VMC66, which possesses a MntH transporter, loss of SsaACB did not significantly alter growth or cellular manganese levels under the same conditions. Interestingly, there were only modest differences between SK36 and its {Delta}ssaACB mutant in competition with Streptococcus mutans in vitro and in a murine oral colonization model. Our results suggest that the heterogeneity of the oral environment may provide a rationale for the variety of manganese transporters found in S. sanguinis and point to strategies for enhancing the safety of oral probiotics. Graphical Abstract O_FIG O_LINKSMALLFIG WIDTH=200 HEIGHT=162 SRC="FIGDIR/small/446192v1_ufig1.gif" ALT="Figure 1"> View larger version (44K): org.highwire.dtl.DTLVardef@a5afa7org.highwire.dtl.DTLVardef@1af3cdforg.highwire.dtl.DTLVardef@1c58693org.highwire.dtl.DTLVardef@1063980_HPS_FORMAT_FIGEXP M_FIG Depiction of methods. Streptococcal strains used are depicted at the top. The four methods illustrated are: 1. S. sanguinis cells were grown in media at pH 7.3 and pH 6.2 and quantified by plating. 2. S. sanguinis cells were grown in a fermentor vessel for RNA-sequencing and metal analysis. 3. S. sanguinis and S. mutans cells were grown in 12-well plates singly or in competition, then plated and the pH of the media measured. 4. S. sanguinis and S. mutans cells were inoculated into the mouths of mice. Oral swabs and dental biofilms from molars were assayed for bacterial composition by qPCR. (Biorender) C_FIG

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Puccio, T., Schultz, A. C., Lizarraga, C. A., Bryant, A. S., Culp, D. J., Burne, R., Kitten, T.. 2021-05-28. Manganese transport by Streptococcus sanguinis in acidic conditions and its impact on growth in vitro and in vivo. https://doi.org/10.1101/2021.05.28.446192

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