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Clostridium difficile has two parallel and essential Sec secretion systems.

Robert P Fagan1, Neil F Fairweather

  • 1Division of Cell and Molecular Biology, Centre for Molecular Microbiology and Infection, Imperial College London, London SW7 2AZ, United Kingdom. r.fagan@imperial.ac.uk

The Journal of Biological Chemistry
|June 11, 2011
PubMed
Summary
This summary is machine-generated.

Clostridium difficile utilizes two Sec systems for protein export. Both SecA1 and SecA2 systems are essential for bacterial growth and the proper translocation of cell wall proteins.

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Area of Science:

  • Microbiology
  • Bacterial Physiology
  • Protein Secretion

Background:

  • Protein translocation across the bacterial cytoplasmic membrane is vital, primarily mediated by the SecYEG channel and SecA ATPase.
  • A second, accessory Sec system involving SecA2 has been identified in some gram-positive bacteria for translocating specific substrates.
  • Clostridium difficile, a pathogenic gram-positive species, possesses two SecA homologs, suggesting the presence of both Sec systems.

Purpose of the Study:

  • To characterize the accessory Sec system in Clostridium difficile.
  • To identify the major substrates translocated by the C. difficile accessory Sec system.
  • To determine the essentiality of both Sec systems for C. difficile growth.

Main Methods:

  • Utilized inducible antisense RNA expression to knockdown secA1 and secA2.
  • Employed dominant-negative alleles of secA1 and secA2 to inhibit Sec system function.
  • Analyzed the translocation of S-layer proteins (SLPs) and cell wall protein CwpV.
  • Quantified protein precursor accumulation and mature protein levels in the cell wall.

Main Results:

  • Export of S-layer proteins (SLPs) and CwpV is dependent on the SecA2 accessory system.
  • Inhibition of either SecA1 or SecA2 led to cytoplasmic accumulation of protein precursors and reduced mature SLPs in the cell wall.
  • Disruption of either SecA1 or SecA2 function via dominant-negative alleles or antisense RNA significantly impaired bacterial growth.

Conclusions:

  • The SecA2 accessory Sec system plays a crucial role in the export of specific cell wall proteins in Clostridium difficile.
  • Both the primary (SecA1) and accessory (SecA2) Sec systems are essential for the viability and growth of C. difficile.
  • Understanding these Sec systems provides insights into the pathogenicity and cell envelope biogenesis of C. difficile.