GpsB regulatory protein and StkP protein kinase have been proposed as molecular switches that balance septal and peripheral (side-wall like) peptidoglycan (PG) synthesis in Streptococcus pneumoniae (pneumococcus); yet, mechanisms of this switching remain unknown. We report that DdivIVA mutations are not epistatic to DgpsB division-protein mutations in progenitor D39 and related genetic backgrounds; nor is GpsB required for StkP localization or FDAA labeling at septal division rings. However, we confirm that reduction of GpsB amount leads to decreased protein phosphorylation by StkP and report that the essentiality of DgpsB mutations is suppressed by inactivation of PhpP protein phosphatase, which concomitantly restores protein phosphorylation levels. DgpsB mutations are also suppressed by other classes of mutations, including one that eliminates protein phosphorylation and may alter division. Moreover, DgpsB mutations are synthetically lethal with Dpbp1a, but not Dpbp2a or Dpbp1b mutations, suggesting GpsB activation of PBP2a activity. Consistent with this result, co-IP experiments showed that GpsB complexes with EzrA, StkP, PBP2a, PBP2b and MreC in pneumococcal cells. Furthermore, depletion of GpsB prevents PBP2x migration to septal centers. These results support a model in which GpsB negatively regulates peripheral PG synthesis by PBP2b and positively regulates septal ring closure through its interactions with StkP-PBP2x.

GpsB regulatory protein and StkP protein kinase have been proposed as molecular switches that balance septal and peripheral (side-wall like) peptidoglycan (PG) synthesis in Streptococcus pneumoniae (pneumococcus); yet, mechanisms of this switching remain unknown. We report that ΔdivIVA mutations are not epistatic to ΔgpsB division-protein mutations in progenitor D39 and related genetic backgrounds; nor is GpsB required for StkP localization or FDAA labeling at septal division rings. However, we confirm that reduction of GpsB amount leads to decreased protein phosphorylation by StkP and report that the essentiality of ΔgpsB mutations is suppressed by inactivation of PhpP protein phosphatase, which concomitantly restores protein phosphorylation levels. ΔgpsB mutations are also suppressed by other classes of mutations, including one that eliminates protein phosphorylation and may alter division. Moreover, ΔgpsB mutations are synthetically lethal with Δpbp1a, but not Δpbp2a or Δpbp1b, mutations, suggesting GpsB activation of PBP2a activity. Consistent with this result, co-IP experiments showed that GpsB complexes with EzrA, StkP, PBP2a, PBP2b, and MreC in pneumococcal cells. Furthermore, depletion of GpsB prevents PBP2x migration to septal centers. These results support a model in which GpsB negatively regulates peripheral PG synthesis by PBP2b and positively

Suppression and Synthetic-Lethal Genetic Relationships of ΔgpsB Mutations Indicate That GpsB Mediates Protein Phosphorylation and Penicillin-Binding Protein Interactions in Streptococcus pneumoniae D39 / Rued Britta, E.; Zheng Jiaqi, J.; Mura, Andrea; Tsui Ho Ching, T.; Boersma Michael, J.; Mazny Jeffrey, L.; Corona, Federico; Perez Amilcar, J.; Fadda, Daniela; Doubravová, Linda; Buriánková, Karolína; Branny, Pavel; Massidda, Orietta; Winkler Malcolm, E.. - In: MOLECULAR MICROBIOLOGY. - ISSN 0950-382X. - 103:6(2017), pp. 931-957. [10.1111/mmi.13613]

Suppression and Synthetic-Lethal Genetic Relationships of ΔgpsB Mutations Indicate That GpsB Mediates Protein Phosphorylation and Penicillin-Binding Protein Interactions in Streptococcus pneumoniae D39

Massidda, Orietta;
2017-01-01

Abstract

GpsB regulatory protein and StkP protein kinase have been proposed as molecular switches that balance septal and peripheral (side-wall like) peptidoglycan (PG) synthesis in Streptococcus pneumoniae (pneumococcus); yet, mechanisms of this switching remain unknown. We report that ΔdivIVA mutations are not epistatic to ΔgpsB division-protein mutations in progenitor D39 and related genetic backgrounds; nor is GpsB required for StkP localization or FDAA labeling at septal division rings. However, we confirm that reduction of GpsB amount leads to decreased protein phosphorylation by StkP and report that the essentiality of ΔgpsB mutations is suppressed by inactivation of PhpP protein phosphatase, which concomitantly restores protein phosphorylation levels. ΔgpsB mutations are also suppressed by other classes of mutations, including one that eliminates protein phosphorylation and may alter division. Moreover, ΔgpsB mutations are synthetically lethal with Δpbp1a, but not Δpbp2a or Δpbp1b, mutations, suggesting GpsB activation of PBP2a activity. Consistent with this result, co-IP experiments showed that GpsB complexes with EzrA, StkP, PBP2a, PBP2b, and MreC in pneumococcal cells. Furthermore, depletion of GpsB prevents PBP2x migration to septal centers. These results support a model in which GpsB negatively regulates peripheral PG synthesis by PBP2b and positively
2017
6
Rued Britta, E.; Zheng Jiaqi, J.; Mura, Andrea; Tsui Ho Ching, T.; Boersma Michael, J.; Mazny Jeffrey, L.; Corona, Federico; Perez Amilcar, J.; Fadda, Daniela; Doubravová, Linda; Buriánková, Karolína; Branny, Pavel; Massidda, Orietta; Winkler Malcolm, E.
Suppression and Synthetic-Lethal Genetic Relationships of ΔgpsB Mutations Indicate That GpsB Mediates Protein Phosphorylation and Penicillin-Binding Protein Interactions in Streptococcus pneumoniae D39 / Rued Britta, E.; Zheng Jiaqi, J.; Mura, Andrea; Tsui Ho Ching, T.; Boersma Michael, J.; Mazny Jeffrey, L.; Corona, Federico; Perez Amilcar, J.; Fadda, Daniela; Doubravová, Linda; Buriánková, Karolína; Branny, Pavel; Massidda, Orietta; Winkler Malcolm, E.. - In: MOLECULAR MICROBIOLOGY. - ISSN 0950-382X. - 103:6(2017), pp. 931-957. [10.1111/mmi.13613]
File in questo prodotto:
File Dimensione Formato  
Rued_et_al-2017-Molecular_Microbiology .pdf

Solo gestori archivio

Descrizione: articolo originale
Tipologia: Versione editoriale (Publisher’s layout)
Licenza: Tutti i diritti riservati (All rights reserved)
Dimensione 1.4 MB
Formato Adobe PDF
1.4 MB Adobe PDF   Visualizza/Apri
Rued_et_al-suppinfo .pdf

accesso aperto

Descrizione: articolo materiale supplementare
Tipologia: Altro materiale allegato (Other attachments)
Licenza: Tutti i diritti riservati (All rights reserved)
Dimensione 2.26 MB
Formato Adobe PDF
2.26 MB Adobe PDF Visualizza/Apri
Lewis-2017-Molecular_Microbiology.pdf

Solo gestori archivio

Descrizione: Journal higlight su richiesta dell'Editor per evidenziare i nostri risultati pubblicatti e l'impatto del lavoro
Tipologia: Versione editoriale (Publisher’s layout)
Licenza: Tutti i diritti riservati (All rights reserved)
Dimensione 220.74 kB
Formato Adobe PDF
220.74 kB Adobe PDF   Visualizza/Apri
Suppression_and_Synthetic-Lethal_Genetic_Relations (1).pdf

accesso aperto

Tipologia: Post-print referato (Refereed author’s manuscript)
Licenza: Tutti i diritti riservati (All rights reserved)
Dimensione 4.57 MB
Formato Adobe PDF
4.57 MB Adobe PDF Visualizza/Apri

I documenti in IRIS sono protetti da copyright e tutti i diritti sono riservati, salvo diversa indicazione

Utilizza questo identificativo per citare o creare un link a questo documento: https://hdl.handle.net/11572/187554
Citazioni
  • ???jsp.display-item.citation.pmc??? 35
  • Scopus 48
  • ???jsp.display-item.citation.isi??? 45
social impact