Penicillin-binding Protein 2x of Streptococcus Pneumoniae: the Mutation Ala707Asp Within the C-terminal PASTA2 Domain Leads to Destabilization

Overview

Journal Microb Drug Resist

Publisher Mary Ann Liebert

Specialties Infectious Diseases
Microbiology

Date 2014 May 21

PMID 24841912

Citations 8

Authors

Inga Schweizer

Katharina Peters

Christoph Stahlmann

Regine Hakenbeck

Dalia Denapaite

Affiliations

Soon will be listed here.

Abstract

Streptococcus pneumoniae penicillin-binding protein 2x (PBP2x) is an enzyme involved in the last stages of peptidoglycan assembly and essential for bacterial growth and survival. PBP2x localizes to the division site, a process that depends on its Penicillin-Binding Protein And Serine-Threonine-kinase Associated (PASTA) domains, which was previously demonstrated via GFP-PBP2x in living cells. During this study a mutant strain was isolated in which the GFP-PBP2x fusion protein did not localize at division sites and it contained reduced amounts of the full-length GFP-PBP2x. We now show that this defect is due to a point mutation within the C-terminal PASTA2 domain of PBP2x. The mutant protein was analyzed in detail in terms of beta-lactam binding, functionality, and localization in live cells. We demonstrate that the mutation affects the GFP-tagged PBP2x variant severely and renders it susceptible to the protease/chaperone HtrA.

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References

Salles C, Creancier L, Claverys J, Mejean V . The high level streptomycin resistance gene from Streptococcus pneumoniae is a homologue of the ribosomal protein S12 gene from Escherichia coli. Nucleic Acids Res. 1992; 20(22):6103. PMC: 334482. DOI: 10.1093/nar/20.22.6103. View

Shah I, Laaberki M, Popham D, Dworkin J . A eukaryotic-like Ser/Thr kinase signals bacteria to exit dormancy in response to peptidoglycan fragments. Cell. 2008; 135(3):486-96. PMC: 2892110. DOI: 10.1016/j.cell.2008.08.039. View

Macheboeuf P, Contreras-Martel C, Job V, Dideberg O, Dessen A . Penicillin binding proteins: key players in bacterial cell cycle and drug resistance processes. FEMS Microbiol Rev. 2006; 30(5):673-91. DOI: 10.1111/j.1574-6976.2006.00024.x. View

Zapun A, Vernet T, Pinho M . The different shapes of cocci. FEMS Microbiol Rev. 2008; 32(2):345-60. DOI: 10.1111/j.1574-6976.2007.00098.x. View

Peters K, Schweizer I, Beilharz K, Stahlmann C, Veening J, Hakenbeck R . Streptococcus pneumoniae PBP2x mid-cell localization requires the C-terminal PASTA domains and is essential for cell shape maintenance. Mol Microbiol. 2014; 92(4):733-55. DOI: 10.1111/mmi.12588. View

Pares S, Mouz N, Petillot Y, Hakenbeck R, Dideberg O . X-ray structure of Streptococcus pneumoniae PBP2x, a primary penicillin target enzyme. Nat Struct Biol. 1996; 3(3):284-9. DOI: 10.1038/nsb0396-284. View

Tsui H, Keen S, Sham L, Wayne K, Winkler M . Dynamic distribution of the SecA and SecY translocase subunits and septal localization of the HtrA surface chaperone/protease during Streptococcus pneumoniae D39 cell division. mBio. 2011; 2(5). PMC: 3188284. DOI: 10.1128/mBio.00202-11. View

Massidda O, Novakova L, Vollmer W . From models to pathogens: how much have we learned about Streptococcus pneumoniae cell division?. Environ Microbiol. 2013; 15(12):3133-57. DOI: 10.1111/1462-2920.12189. View

Maurer P, Koch B, Zerfass I, Krauss J, van der Linden M, Frere J . Penicillin-binding protein 2x of Streptococcus pneumoniae: three new mutational pathways for remodelling an essential enzyme into a resistance determinant. J Mol Biol. 2008; 376(5):1403-16. DOI: 10.1016/j.jmb.2007.12.058. View

10.

Lanie J, Ng W, Kazmierczak K, Andrzejewski T, Davidsen T, Wayne K . Genome sequence of Avery's virulent serotype 2 strain D39 of Streptococcus pneumoniae and comparison with that of unencapsulated laboratory strain R6. J Bacteriol. 2006; 189(1):38-51. PMC: 1797212. DOI: 10.1128/JB.01148-06. View

11.

Alloing G, Granadel C, Morrison D, Claverys J . Competence pheromone, oligopeptide permease, and induction of competence in Streptococcus pneumoniae. Mol Microbiol. 1996; 21(3):471-8. DOI: 10.1111/j.1365-2958.1996.tb02556.x. View

12.

Zapun A, Contreras-Martel C, Vernet T . Penicillin-binding proteins and beta-lactam resistance. FEMS Microbiol Rev. 2008; 32(2):361-85. DOI: 10.1111/j.1574-6976.2007.00095.x. View

13.

Mascher T, Heintz M, Zahner D, Merai M, Hakenbeck R . The CiaRH system of Streptococcus pneumoniae prevents lysis during stress induced by treatment with cell wall inhibitors and by mutations in pbp2x involved in beta-lactam resistance. J Bacteriol. 2006; 188(5):1959-68. PMC: 1426552. DOI: 10.1128/JB.188.5.1959-1968.2006. View

14.

Kell C, Sharma U, Dowson C, Town C, Balganesh T, Spratt B . Deletion analysis of the essentiality of penicillin-binding proteins 1A, 2B and 2X of Streptococcus pneumoniae. FEMS Microbiol Lett. 1993; 106(2):171-5. DOI: 10.1111/j.1574-6968.1993.tb05954.x. View

15.

Maurer P, Todorova K, Sauerbier J, Hakenbeck R . Mutations in Streptococcus pneumoniae penicillin-binding protein 2x: importance of the C-terminal penicillin-binding protein and serine/threonine kinase-associated domains for beta-lactam binding. Microb Drug Resist. 2012; 18(3):314-21. DOI: 10.1089/mdr.2012.0022. View

16.

Dessen A, Mouz N, Gordon E, Hopkins J, Dideberg O . Crystal structure of PBP2x from a highly penicillin-resistant Streptococcus pneumoniae clinical isolate: a mosaic framework containing 83 mutations. J Biol Chem. 2001; 276(48):45106-12. DOI: 10.1074/jbc.M107608200. View

17.

Lacks S, HOTCHKISS R . A study of the genetic material determining an enzyme in Pneumococcus. Biochim Biophys Acta. 1960; 39:508-18. DOI: 10.1016/0006-3002(60)90205-5. View

18.

Mir M, Asong J, Li X, Cardot J, Boons G, Husson R . The extracytoplasmic domain of the Mycobacterium tuberculosis Ser/Thr kinase PknB binds specific muropeptides and is required for PknB localization. PLoS Pathog. 2011; 7(7):e1002182. PMC: 3145798. DOI: 10.1371/journal.ppat.1002182. View

19.

OTTOLENGHI E, HOTCHKISS R . Release of genetic transforming agent from pneumococcal cultures during growth and disintegration. J Exp Med. 1962; 116:491-519. PMC: 2137623. DOI: 10.1084/jem.116.4.491. View

20.

Morlot C, Bayle L, Jacq M, Fleurie A, Tourcier G, Galisson F . Interaction of Penicillin-Binding Protein 2x and Ser/Thr protein kinase StkP, two key players in Streptococcus pneumoniae R6 morphogenesis. Mol Microbiol. 2013; 90(1):88-102. DOI: 10.1111/mmi.12348. View