The effects of methionine acquisition and synthesis on Streptococcus pneumoniae growth and virulence

2.50
Hdl Handle:
http://hdl.handle.net/10547/576993
Title:
The effects of methionine acquisition and synthesis on Streptococcus pneumoniae growth and virulence
Authors:
Basavanna, Shilpa; Chimalapati, Suneeta; Rubbo, Bruna; Yuste, Jose; Hosie, Arthur H.F. ( 0000-0002-1327-7901 ) ; Thomas, Gavin H.; Brown, Jeremy S.; Maqbool, Abbas; Ogunniyi, Abiodun D.; Paton, James C.
Abstract:
Bacterial pathogens need to acquire nutrients from the host, but for many nutrients their importance during infection remain poorly understood. We have investigated the importance of methionine acquisition and synthesis for Streptococcus pneumoniae growth and virulence using strains with gene deletions affecting a putative methionine ABC transporter lipoprotein (Sp_0149, metQ) and/or methionine biosynthesis enzymes (Sp_0585 - Sp_0586, metE and metF). Immunoblot analysis confirmed MetQ was a lipoprotein and present in all S. pneumoniae strains investigated. However, vaccination with MetQ did not prevent fatal S. pneumoniae infection in mice despite stimulating a strong specific IgG response. Tryptophan fluorescence spectroscopy and isothermal titration calorimetry demonstrated that MetQ has both a high affinity and specificity for L-methionine with a KD of ~25 nM, and a ΔmetQ strain had reduced uptake of C14-methionine. Growth of the ΔmetQ/ΔmetEF strain was greatly impaired in chemically defined medium containing low concentrations of methionine and in blood but was partially restored by addition of high concentrations of exogenous methionine. Mixed infection models showed no attenuation of the ΔmetQ, ΔmetEF and ΔmetQ/ΔmetEF strains in their ability to colonise the mouse nasopharnyx. In a mouse model of systemic infection although significant infection was established in all mice, there were reduced spleen bacterial CFU after infection with the ΔmetQ/ΔmetEF strain compared to the wild-type strain. These data demonstrate that Sp_0149 encodes a high affinity methionine ABC transporter lipoprotein and that Sp_0585 – Sp_0586 are likely to be required for methionine synthesis. Although Sp_0149 and Sp_0585-Sp_0586 make a contribution towards full virulence, neither was essential for S. pneumoniae survival during infection.
Citation:
Basavanna, S., Chimalapati, S., Rubbo, B., Yuste, J., Hosie, A., Thomas, G., Brown, J.S., (2013) 'The effects of methionine acquisition and synthesis on Streptococcus pneumoniae growth and virulence'. PLOS One 8 (1)
Publisher:
PLoS
Journal:
PLoS ONE
Issue Date:
Jan-2013
URI:
http://hdl.handle.net/10547/576993
DOI:
10.1371/journal.pone.0049638
Additional Links:
http://journals.plos.org/plosone/article?id=10.1371/journal.pone.0049638
Type:
Article
Language:
en
ISSN:
1932-6203
Sponsors:
This work was undertaken at UCLH/UCL, which received a proportion of funding from the Department of Health’s NIHR Biomedical Research Centre’s funding scheme, and was supported by the UCL Charities funding and grants from the British Lung Foundation (P05/3) and the Wellcome Trust (grant reference 076442). The funders had no role in study design, data collection and analysis, decision to publish, or preparation of the manuscript.
Appears in Collections:
Cell and Cryobiology Research Group

Full metadata record

DC FieldValue Language
dc.contributor.authorBasavanna, Shilpaen
dc.contributor.authorChimalapati, Suneetaen
dc.contributor.authorRubbo, Brunaen
dc.contributor.authorYuste, Joseen
dc.contributor.authorHosie, Arthur H.F.en
dc.contributor.authorThomas, Gavin H.en
dc.contributor.authorBrown, Jeremy S.en
dc.contributor.authorMaqbool, Abbasen
dc.contributor.authorOgunniyi, Abiodun D.en
dc.contributor.authorPaton, James C.en
dc.date.accessioned2015-09-09T10:09:20Zen
dc.date.available2015-09-09T10:09:20Zen
dc.date.issued2013-01en
dc.identifier.citationBasavanna, S., Chimalapati, S., Rubbo, B., Yuste, J., Hosie, A., Thomas, G., Brown, J.S., (2013) 'The effects of methionine acquisition and synthesis on Streptococcus pneumoniae growth and virulence'. PLOS One 8 (1)en
dc.identifier.issn1932-6203en
dc.identifier.doi10.1371/journal.pone.0049638en
dc.identifier.urihttp://hdl.handle.net/10547/576993en
dc.description.abstractBacterial pathogens need to acquire nutrients from the host, but for many nutrients their importance during infection remain poorly understood. We have investigated the importance of methionine acquisition and synthesis for Streptococcus pneumoniae growth and virulence using strains with gene deletions affecting a putative methionine ABC transporter lipoprotein (Sp_0149, metQ) and/or methionine biosynthesis enzymes (Sp_0585 - Sp_0586, metE and metF). Immunoblot analysis confirmed MetQ was a lipoprotein and present in all S. pneumoniae strains investigated. However, vaccination with MetQ did not prevent fatal S. pneumoniae infection in mice despite stimulating a strong specific IgG response. Tryptophan fluorescence spectroscopy and isothermal titration calorimetry demonstrated that MetQ has both a high affinity and specificity for L-methionine with a KD of ~25 nM, and a ΔmetQ strain had reduced uptake of C14-methionine. Growth of the ΔmetQ/ΔmetEF strain was greatly impaired in chemically defined medium containing low concentrations of methionine and in blood but was partially restored by addition of high concentrations of exogenous methionine. Mixed infection models showed no attenuation of the ΔmetQ, ΔmetEF and ΔmetQ/ΔmetEF strains in their ability to colonise the mouse nasopharnyx. In a mouse model of systemic infection although significant infection was established in all mice, there were reduced spleen bacterial CFU after infection with the ΔmetQ/ΔmetEF strain compared to the wild-type strain. These data demonstrate that Sp_0149 encodes a high affinity methionine ABC transporter lipoprotein and that Sp_0585 – Sp_0586 are likely to be required for methionine synthesis. Although Sp_0149 and Sp_0585-Sp_0586 make a contribution towards full virulence, neither was essential for S. pneumoniae survival during infection.en
dc.description.sponsorshipThis work was undertaken at UCLH/UCL, which received a proportion of funding from the Department of Health’s NIHR Biomedical Research Centre’s funding scheme, and was supported by the UCL Charities funding and grants from the British Lung Foundation (P05/3) and the Wellcome Trust (grant reference 076442). The funders had no role in study design, data collection and analysis, decision to publish, or preparation of the manuscript.en
dc.language.isoenen
dc.publisherPLoSen
dc.relation.urlhttp://journals.plos.org/plosone/article?id=10.1371/journal.pone.0049638en
dc.subjectmethionineen
dc.subjectStreptococcus pneumoniaeen
dc.titleThe effects of methionine acquisition and synthesis on Streptococcus pneumoniae growth and virulenceen
dc.typeArticleen
dc.identifier.journalPLoS ONEen
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