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RepD-mediated recruitment of PcrA helicase at the Staphylococcus aureus pC221 plasmid replication origin, oriD

By Cristina Machόn, N.H. Thomson, G.P. Lynch, David J. Scott, C.D. Thomas and Panos Soultanas


Plasmid encoded replication initiation (Rep) proteins recruit host helicases to plasmid replication origins. Previously, we showed that RepD recruits directionally the PcrA helicase to the pC221 oriD, remains associated with it, and increases its processivity during plasmid unwinding. Here we show that RepD forms a complex extending upstream and downstream of the core oriD. Binding of RepD causes remodelling of a region upstream from the core oriD forming a 'landing pad' for the PcrA. PcrA is recruited by this extended RepD-DNA complex via an interaction with RepD at this upstream site. PcrA appears to have weak affinity for this region even in the absence of RepD. Upon binding of ADPNP (non-hydrolysable analogue of ATP), by PcrA, a conformational rearrangement of the RepD-PcrA-ATP initiation complex confines it strictly within the boundaries of the core oriD. We conclude that RepD-mediated recruitment of PcrA at oriD is a three step process. First, an extended RepD-oriD complex includes a region upstream from the core oriD; second, the PcrA is recruited to this upstream region and thirdly upon ATP-binding PcrA relocates within the core oriD

Publisher: Oxford Journals
Year: 2010
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Provided by: Nottingham ePrints

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  3. (1993). Characterization of the Staphylococcus aureus chromosomal gene pcrA, identified by mutations affecting plasmid pT181 replication. doi
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  6. (2004). DNA footprinting and biophysical characterization of the controller protein C.Ahdl suggests the basis of a genetic switch.
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  8. (1994). Footprinting with Exonuclease III.
  9. (2006). Genetic and biochemical characterization of the Streptococcus pneumoniae PcrA helicase and its role in plasmid rolling circle replication.
  10. (1990). In vitro studies of the initiation of staphylococcal plasmid replication. Specificity of RepD for its origin (oriD), and characterization of the Rep-ori tyrosyl ester intermediate.
  11. (2006). Investigating the basis of substrate recognition in the pC221 relaxosome.
  12. (1994). Multiple DNA conformational changes induced by an initiator protein precede the nicking reaction in a rolling circle replication origin.
  13. (2009). PcrA helicase tightly couples ATP hydrolysis to unwinding double-stranded DNA, modulated by the initiator protein for plasmid replication, doi
  14. (1998). PcrA is an essential DNA helicase of Bacillus subtilis fulfilling functions both in repair and rolling-circle replication. doi
  15. (1993). Plasmid pT181-linked suppressors of the Staphylococcus aureus pcrA3 chromosomal mutation. doi
  16. (1999). Plasmid replication initiator protein RepD increases the processivity of PcrA DNA helicase. doi
  17. (2005). Plasmid rolling-circle replication: highlights of two decades of research.
  18. (2000). Plasmid rolling-circle replication: recent developments.
  19. (2009). Plasmid-mediated antibiotic resistance in Staphylococcus aureus from patients and non-patients.
  20. (1998). Replication and control of circular bacterial plasmids.
  21. (1988). Replication termination for staphylococcal plasmids: plasmids pT181 and pC221 cross-react in the termination process.
  22. (2000). Size distribution analysis of macromolecules by sedimentation velocity ultracentrifugation and Lamm equation modeling.
  23. (1990). Specificity of RepC protein in plasmid pT181 DNA replication.
  24. (1989). Staphylococcal plasmids and their replication.
  25. (1989). Staphylococcus aureus chromosomal mutations that decrease efficiency of Rep utilization in replication of pT181 and related plasmids.
  26. (1986). Static and initiator protein-enhanced bending of DNA at a replication origin.
  27. (1988). Structural relationships among chloramphenicol-resistance plasmids of Staphylococcus aureus.
  28. (1989). Synthesis of single-stranded plasmid pT181 DNA in vitro: initiation and termination of DNA replication.
  29. (2009). The ATPase cycle of PcrA helicase and its coupling to translocation on DNA.
  30. (1985). The replication initiator protein of plasmid pT181 has sequence-specific endonuclease and topoisomerase-like activities.
  31. (1991). The Staphylococcus aureus mutation pcrA3 leads to the accumulation of pT181 replication initiation complexes. doi

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