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An Assay for Quantifying Protein-RNA Binding in Bacteria
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Published on: June 12, 2019

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An outer membrane determinant for RNA phage genome entry in Pseudomonas aeruginosa

Hee-Won Bae1, Shin-Yae Choi1, You-Hee Cho1

  • 1Department of Pharmacy, College of Pharmacy and Institute of Pharmaceutical Sciences, CHA University, Gyeonggi-do 13488, Korea.

Iscience
|January 12, 2024
PubMed

Insights

RNA phage PP7 infectivity in Pseudomonas aeruginosa is limited by lipopolysaccharide (LPS) structure, not just type IV pilus (TFP) proteins. Modifying LPS biosynthesis enhances phage entry, revealing new host range determinants.

Area of Science:

  • Microbiology
  • Virology
  • Bacteriology

Background:

  • Phage host range is influenced by multiple infection stages.
  • Previous studies identified type IV pilus (TFP) as a receptor for RNA phage PP7 in Pseudomonas aeruginosa.
  • Variations in TFP genes between P. aeruginosa strains (PAO1 vs. PA14) affect PP7 susceptibility.

Purpose of the Study:

  • To investigate additional determinants beyond TFP that limit PP7 phage infectivity in P. aeruginosa.
  • To explore the role of lipopolysaccharide (LPS) structure in regulating PP7 phage entry.

Main Methods:

  • Created a PA14 derivative (PA14P) expressing PAO1 pilin gene.
  • Conducted transposon mutant screens to identify genes affecting PP7 infectivity in PA14P.
  • Utilized Polymyxin B treatment to assess its effect on phage entry.

Main Results:

  • PP7 infectivity was restored in PA14P mutants lacking htrB2, a gene involved in lipid A biosynthesis.
  • The absence of the secondary acyltransferase (HtrB2) led to a loosened LPS structure.
  • This altered LPS structure facilitated increased RNA phage entry.
  • Polymyxin B treatment also selectively enhanced RNA phage entry.

Conclusions:

  • Lipopolysaccharide (LPS) structure is a significant determinant limiting the entry stage of RNA phages.
  • Factors beyond TFP, specifically LPS modifications, contribute to the host range of PP7 in P. aeruginosa.
  • Targeting LPS biosynthesis or structure could be a strategy to modulate phage-bacterial interactions.
Keywords:
BiochemistryViral microbiologymicrobiologymolecular microbiology

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