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The effect of rhamnolipid production on Pseudomonas aeruginosa physiology

Başlık çevirisi mevcut değil.

  1. Tez No: 402935
  2. Yazar: ŞEBNEM BUKAVAZ
  3. Danışmanlar: DR. FRANK ROSENAU
  4. Tez Türü: Doktora
  5. Konular: Moleküler Tıp, Molecular Medicine
  6. Anahtar Kelimeler: Belirtilmemiş.
  7. Yıl: 2015
  8. Dil: İngilizce
  9. Üniversite: Ulm Unıversıty
  10. Enstitü: Yurtdışı Enstitü
  11. Ana Bilim Dalı: Belirtilmemiş.
  12. Bilim Dalı: Belirtilmemiş.
  13. Sayfa Sayısı: Belirtilmemiş.

Özet

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Özet (Çeviri)

Pseudomonas aeruginosa is a notorious human pathogen associated with a range of lifethreatening nosocomial infections. P. aeruginosa is known to produce a range of virulence factors that enhance its ability to damage the host tissue and cause disease. One of the most important virulent-related molecules is rhamnolipid. The rhamnolipids are known in P. aeruginosa as a multifunctional surfactant. Identifying the functions of rhamnolipids in detail is clearly warranted to elucidate of the mechanisms of P. aeruginosa pathogenesis and to explore new strategies for treatment. P. aeruginosa regulates rhamnolipid production by using an intercellular communication mechanism called quorum sensing, which is mediated by small signaling molecules, or auto-inducers. In the present study, two rhamnolipid mutants were constructed and their physiological functions on P. aeruginosa were investigated under Pi depletion conditions. In this study, the surface-associated GroEL protein, which can be a host-pathogen interaction and anti-biofilm target, promoted early biofilm formation in P. aeruginosa. Pi depletion is known to rapidly develop following major surgery and organ injury and independently predicts the development of lethal sepsis (261),(106). The QSSM profile identification study for rhamnolipid-deficient mutants revealed high PQS and C4-HSL levels that significantly impact several virulence-associated phenotypes, such as Pvds-regulated endoprotease, lipase and phospholipase production, which was confirmed by extracellular proteomic and phenotypic analyses. However, similar phenotypic characteristics for a mono-rhamnolipid-producing strain and wild type PAO1 were obtained. The non-swarming and flat biofilm forming phenotype of the ΔrhlA strain suggested an altered outer membrane protein profile, which was verified by proteomic analyses. Furthermore, the GroEL protein was detected at different expression levels in a rhamnolipiddeficient mutant compared to wild type PAO1. This result led to the question of whether GroEL has an additional function, and GroEL was successfully overexpressed in a rhamnolipiddeficient mutant for subsequent functional characterization. The higher expression level of extracellular surface-associated GroEL protein was confirmed by extracellular and cytoplasmic proteome analyses. The suggested role was shown by crystal violet assay and the control of cell lysis was confirmed by β-lactamase activity assay and LipH Western blotting analyses. The presented data in this study is the first report of a proteome-based study of rhamnolipid mutants and the surface-associated GroEL and its new role in P. aeruginosa. Moreover, these results indicate anti-biofilm targets in P. aeruginosa.

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