Competition of bacteriophage polypeptides with native replicase proteins for binding to the DNA sliding clamp reveals a novel mechanism for DNA replication arrest in Staphylococcus aureus

Adam Belley, Mario Callejo, Francis Arhin, Mohammed Dehbi, Ibtihal Fadhil, Jing Liu, Geoffrey McKay, Ramakrishnan Srikumar, Pascale Bauda, Nhuan Ha, Michael DuBow, Philippe Gros, Jerry Pelletier, Greg Moeck

Research output: Contribution to journalArticle

15 Citations (Scopus)

Abstract

Bacteriophages have evolved specific mechanisms that redirect bacterial metabolic pathways to the bacteriophage reproduction cycle. In this study, we characterized the bactericidal mechanism of two polypeptides from bacteriophages Twort and G1 that target the DNA sliding clamp of Staphylococcus aureus. The DNA sliding clamp, which tethers DNA polymerase to its template and thereby confers processivity upon the enzyme, was found to be essential for the viability of S. aureus. Expression of polypeptides TwortORF168 and G1ORF240 in S. aureus selectively inhibited DNA replication which in turn resulted in cell death. Both polypeptides specifically inhibited the S. aureus DNA replicase that was reconstituted in vitro but not the corresponding replicase of Streptococcus pyogenes. We demonstrated that inhibition of DNA synthesis is multifaceted and occurs via binding the DNA sliding clamp: TwortORF168 and G1ORF240 bound tightly to the DNA sliding clamp and prevented both its loading onto DNA and its interaction with DNA polymerase C. These results elucidate the impact of bacteriophage polypeptide expression upon DNA replication in the growing cell.

Original languageEnglish
Pages (from-to)1132-1143
Number of pages12
JournalMolecular Microbiology
Volume62
Issue number4
DOIs
Publication statusPublished - 1 Nov 2006
Externally publishedYes

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DNA-Binding Proteins
DNA Replication
Bacteriophages
Staphylococcus aureus
Peptides
DNA
Streptococcus pyogenes
DNA-Directed DNA Polymerase
Metabolic Networks and Pathways
Reproduction
Cell Death
Enzymes

ASJC Scopus subject areas

  • Molecular Biology
  • Microbiology

Cite this

Competition of bacteriophage polypeptides with native replicase proteins for binding to the DNA sliding clamp reveals a novel mechanism for DNA replication arrest in Staphylococcus aureus. / Belley, Adam; Callejo, Mario; Arhin, Francis; Dehbi, Mohammed; Fadhil, Ibtihal; Liu, Jing; McKay, Geoffrey; Srikumar, Ramakrishnan; Bauda, Pascale; Ha, Nhuan; DuBow, Michael; Gros, Philippe; Pelletier, Jerry; Moeck, Greg.

In: Molecular Microbiology, Vol. 62, No. 4, 01.11.2006, p. 1132-1143.

Research output: Contribution to journalArticle

Belley, A, Callejo, M, Arhin, F, Dehbi, M, Fadhil, I, Liu, J, McKay, G, Srikumar, R, Bauda, P, Ha, N, DuBow, M, Gros, P, Pelletier, J & Moeck, G 2006, 'Competition of bacteriophage polypeptides with native replicase proteins for binding to the DNA sliding clamp reveals a novel mechanism for DNA replication arrest in Staphylococcus aureus', Molecular Microbiology, vol. 62, no. 4, pp. 1132-1143. https://doi.org/10.1111/j.1365-2958.2006.05427.x
Belley, Adam ; Callejo, Mario ; Arhin, Francis ; Dehbi, Mohammed ; Fadhil, Ibtihal ; Liu, Jing ; McKay, Geoffrey ; Srikumar, Ramakrishnan ; Bauda, Pascale ; Ha, Nhuan ; DuBow, Michael ; Gros, Philippe ; Pelletier, Jerry ; Moeck, Greg. / Competition of bacteriophage polypeptides with native replicase proteins for binding to the DNA sliding clamp reveals a novel mechanism for DNA replication arrest in Staphylococcus aureus. In: Molecular Microbiology. 2006 ; Vol. 62, No. 4. pp. 1132-1143.
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