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Related Experiment Videos

Insertion mutations in pilE differentially alter gonococcal pilin antigenic variation.

B Howell-Adams1, H S Seifert

  • 1Department of Microbiology-Immunology, Northwestern University Medical School, Chicago, Illinois 60611, USA.

Journal of Bacteriology
|September 28, 1999
PubMed
Summary

Neisseria gonorrhoeae uses pilus antigenic variation to evade the immune system. Disrupting DNA sequences in the expressed pilin gene (pilE) impairs recombination, suggesting distinct roles for silent (pilS) and expressed (pilE) genes in this process.

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Area of Science:

  • Microbiology
  • Molecular Biology
  • Genetics

Background:

  • Neisseria gonorrhoeae exhibits pilus antigenic variation through DNA recombination.
  • Silent pilin loci (pilS) transfer DNA to the expressed pilin gene (pilE), altering the pilin protein.
  • Previous studies indicated conserved DNA sequences (cys2) are crucial for this recombination.

Purpose of the Study:

  • To investigate the role of the expressed pilin gene (pilE) in pilus antigenic variation.
  • To determine the effect of disrupting the conserved cys2 sequence in pilE on recombination.
  • To compare the functions of silent (pilS) and expressed (pilE) pilin loci in antigenic variation.

Main Methods:

  • Introduction of heterologous DNA insertions and deletions into the pilE gene.

Related Experiment Videos

  • Analysis of pilin recombination frequency and spectrum of utilized donor sequences.
  • Comparison of recombination defects caused by modifications in pilE versus pilS.

    • Main Results:

    • Disruption or deletion of the cys2 sequence in pilE significantly affects pilin recombination.
    • Insertions in pilE impaired, but did not completely block, recombination of flanking sequences.
    • Modifications in pilE altered the repertoire of silent donor copies used during variation.

    Conclusions:

    • The expressed pilin gene (pilE) and silent pilin loci (pilS) play distinct roles in antigenic variation.
    • Disruption of conserved sequences in pilE impacts recombination differently than in pilS.
    • Alternative recombination mechanisms become active when high-frequency pathways are inhibited.