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Volume 157, Issue 10

Insights into the function of protein P30 from orthologous gene replacement Free

Abstract

The attachment organelles of bacterial species belonging to the phylogenetic cluster are required for host cytadherence, gliding motility and virulence. Despite being closely related, these bacteria possess distinct cellular morphologies and gliding characteristics. The molecular mechanisms for most attachment organelle phenotypes, including shape and ability to power motility, are obscure. The attachment organelle-associated P30 protein of is implicated in both adherence and motility, with mutations negatively impacting cell morphology, adherence, gliding and virulence. To test whether the P30 alleles of different mycoplasma species confer species-specific attachment organelle properties, we created an strain in which the P30 orthologue, P32, was substituted for the native P30. Selected clones were visualized by scanning electron microscopy to assess morphology and by indirect immunofluorescence microscopy to localize P32. Cytadherence ability and gliding motility were assessed by haemadsorption assay and phase-contrast microcinematography, respectively. Cell and attachment organelle morphologies were indistinguishable from wild-type as well as II-3 expressing a C-terminally 6×His-tagged P30 construct. P32 was localized to the tip of the attachment organelle of transformant cells. Although a specific role for P30 in species-specific phenotypes was not identified, this first test of orthologous gene replacement in different mycoplasma species demonstrates that the differences in the and proteins contribute little if anything to the different attachment organelle phenotypes between these species.

  • Received:
  • Accepted:
  • Revised:
  • Published Online:
Funding
This study was supported by the:
  • National Institutes of Health (Award R15 AI073994)
  • Miami University
© 2011 SGM
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2011-10-01
2024-04-25
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Insights into the function of Mycoplasma pneumoniae protein P30 from orthologous gene replacement
Microbiology 157, 2862 (2011); https://doi.org/10.1099/mic.0.052464-0
/content/journal/micro/10.1099/mic.0.052464-0
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