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Identification of an additional minor pilin essential for piliation in the archaeon Methanococcus maripaludis

Divya B Nair1, Daniel K C Chung1, James Schneider1

  • 1Department of Biomedical and Molecular Sciences, Queen's University, Kingston, Ontario, Canada.

Plos One
|January 4, 2014
PubMed

Insights

Researchers investigated the function of six pilin genes in Methanococcus maripaludis. Deleting the mmp1283 gene resulted in nonpiliated cells, identifying it as essential for type IV pili assembly in this archaeon.

Area of Science:

  • Microbiology
  • Archaea
  • Cellular Biology

Background:

  • Methanococcus maripaludis possesses archaella and type IV-like pili.
  • MMP1685 is the major structural pilin; EpdA, EpdB, and EpdC are minor pilins.
  • Pilin proteins are processed by the pilin-specific prepilin peptidase EppA.

Purpose of the Study:

  • To determine the essentiality of six previously identified pilin-like genes for pili formation in M. maripaludis.
  • To investigate the role of mmp0528, mmp0600, mmp0601, mmp0709, mmp0903, and mmp1283 in type IV pili assembly.

Main Methods:

  • Gene deletion of six putative pilin-encoding genes in M. maripaludis.
  • Analysis of pili formation in mutant strains using electron microscopy.
  • RT-PCR to confirm mRNA transcript presence.
  • Complementation studies to validate gene function.

Main Results:

  • mRNA transcripts for all six genes were detected.
  • Deletion of mmp1283 resulted in a complete loss of pili formation (nonpiliated cells).
  • Complementation with a wildtype mmp1283 gene restored piliation.

Conclusions:

  • The mmp1283 gene is essential for the assembly of type IV pili in Methanococcus maripaludis.
  • The study highlights the complexity of the type IV pili system in this archaeon.
  • The functions of the other five pilin-like genes and potential alternative pili structures remain to be elucidated.

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