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. 2024 Jun 10;24(1):207.
doi: 10.1186/s12866-024-03349-7.

Quorum sensing gene lasR promotes phage vB_Pae_PLY infection in Pseudomonas aeruginosa

Yan Liu  1 , Zhuocheng Yao  1 , Zhenzhi Mao  1 , Miran Tang  1 , Huanchang Chen  1 , Changrui Qian  1 , Weiliang Zeng  1 , Tieli Zhou  2 , Qing Wu  3
Affiliations

Quorum sensing gene lasR promotes phage vB_Pae_PLY infection in Pseudomonas aeruginosa

Yan Liu et al. BMC Microbiol. .

Abstract

Background: Quorum sensing (QS) is a cell density-based intercellular communication system that controls virulence gene expression and biofilm formation. In Pseudomonas aeruginosa (P. aeruginosa), the LasR system sits at the top of the QS hierarchy and coordinates the expression of a series of important traits. However, the role of lasR in phage infection remains unclear. This study aims to investigate the role of lasR QS in phage infection.

Methods: The P. aeruginosa phage was isolated from sewage, and its biological characteristics and whole genome were analyzed. The adsorption receptor was identified via a phage adsorption assay. Following lasR gene knockout, the adsorption rate and bactericidal activity of phage were analyzed. Finally, real-time quantitative polymerase chain reaction (RT-qPCR) was conducted to explore how lasR promoting phage infection.

Results: The lytic phage vB_Pae_PLY was isolated and lipopolysaccharide (LPS) was identified as its adsorption receptor. The adsorption rate and bactericidal activity of vB_Pae_PLY were reduced after lasR knockout. RT-qPCR results showed that the expression of galU, a key gene involved in LPS synthesis, was down-regulated, and several genes related to type IV pili (T4P) were also down-regulated in the lasR mutant PaΔlasR.

Conclusions: The study showed that QS lasR may promote phage vB_Pae_PLY infection by involving in the synthesis of LPS and T4P. This study provides an example of QS in promoting phage infection and deepens the understanding of phage-bacteria interactions.

Keywords: Pseudomonas aeruginosa; lasR; Phage infection; Phage receptor; Quorum sensing.

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Conflict of interest statement

The authors declare no competing interests.

Figures

Fig. 1
Fig. 1
(a) Plaque formation on a double-layer agar plate of vB_Pae_PLY; (b) Morphology of vB_Pae_PLY under TEM.
Fig. 2
Fig. 2
Lytic activity of vB_Pae_PLY against 40 strains of P. aeruginosa. +, phage formed a clear zone or plaque; −, phage formed no clear zone nor plaque
Fig. 3
Fig. 3
The one-step growth curve showed a latency period of 40 min and high burst size (up to 853 PFU/infected cell)
Fig. 4
Fig. 4
The genomic structure of vB_Pae_PLY. The first circles represent the 64 coding DNA sequences (CDS) of the phage genome. The second circle shows GC content. The third circle shows the GC skew
Fig. 5
Fig. 5
Phage taxonomy and phylogeny analysis. (a) Viral proteomic tree; (b) Phylogenetic tree based on the 349 genome sequences of Pseudomonadota phages of the family Autophagoviridae
Fig. 6
Fig. 6
Identification of LPS as an important absorption receptor of vB_Pae_PLY. (a) NaIO4 treatment significantly reduced the adsorption of vB_Pae_PLY; (b) LPS adsorption assay. The adsorption rate was increased in the LPS-added group compared to that in the control group. **P < 0.01, ns, no significance
Fig. 7
Fig. 7
Phage sensitivity assay. (a) Ten-fold serial dilutions of the phage vB_Pae_PLY plated on PAO1 and lasR mutants PaΔlasR; (b) Comparison of adsorption rates of the phage vB_Pae_PLY to PAO1 and PAO1ΔlasR; (c) Time killing effect in the presence or absence of phage at an MOI of 10. *P < 0.05
Fig. 8
Fig. 8
lasR knockout down-regulated the expression of phage absorption-related genes. The reference gene was rpsL. *P < 0.05, **P < 0.01, and ***P < 0.001
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