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Multiplex PCR Assay for Typing of Staphylococcal Cassette Chromosome Mec Types I to V in Methicillin-resistant Staphylococcus aureus
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Staphylococcal SCCmec elements encode an active MCM-like helicase and thus may be replicative.

Ignacio Mir-Sanchis1, Christina A Roman1, Agnieszka Misiura1

  • 1Department of Biochemistry and Molecular Biology, The University of Chicago, Chicago, Illinois, USA.

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|August 30, 2016
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Summary

Staphylococcal cassette chromosome (SCC) elements, responsible for MRSA, are not nonreplicative. We identified a novel helicase, Cch, and DNA-binding protein, suggesting SCC replication enhances horizontal gene transfer.

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Area of Science:

  • Microbiology
  • Molecular Biology
  • Genomics

Background:

  • Methicillin-resistant Staphylococcus aureus (MRSA) poses a significant global health challenge.
  • The staphylococcal cassette chromosome (SCC) is the mobile genetic element conferring the MRSA phenotype.
  • SCC has been traditionally considered a nonreplicative mobile element.

Purpose of the Study:

  • To investigate the potential DNA replication functions of conserved proteins encoded by SCC.
  • To characterize the Cch protein and its associated functions.
  • To explore the broader evolutionary context of SCC elements within mobile genetic elements.

Main Methods:

  • Bioinformatic analysis to predict protein functions.
  • Biochemical assays to determine helicase activity.
  • X-ray crystallography to elucidate protein structure.
  • Comparative genomics to analyze related mobile elements.

Main Results:

  • The SCC protein Cch exhibits homology to initiator helicases from other genomic islands and possesses active 3'-to-5' helicase activity.
  • An adjacent open reading frame (ORF) encodes a single-stranded DNA-binding protein.
  • The crystal structure reveals Cch forms a hexameric ring and belongs to the AAA+ ATPase superfamily, similar to MCM replicative helicases.
  • SCC elements are part of a larger family of mobile elements that encode replication initiators upstream of recombinases.

Conclusions:

  • SCC elements possess a functional replication initiation system, challenging the long-held view of their nonreplicative nature.
  • The identified replication machinery likely enhances the efficiency of SCC's horizontal gene transfer.
  • This finding provides new insights into the evolution and dissemination of antibiotic resistance genes.