Carpe pili! Hunting strategy of phage JBD30 revealed by
combination of cryo-electron and fluorescent microscopy

a 2023

Carpe pili! Hunting strategy of phage JBD30 revealed by combination of cryo-electron and fluorescent microscopy

VALENTOVÁ, Lucie, Tibor FÜZIK a Pavel PLEVKA

Základní údaje

Originální název

Carpe pili! Hunting strategy of phage JBD30 revealed by combination of cryo-electron and fluorescent microscopy

Autoři

VALENTOVÁ, Lucie, Tibor FÜZIK a Pavel PLEVKA

Vydání

XVIII Discussions in Structural Molecular Biology and 5th User Meeting of CIISB, 2023

Další údaje

Jazyk

angličtina

Typ výsledku

Konferenční abstrakt

Obor

10607 Virology

Stát vydavatele

Česká republika

Utajení

není předmětem státního či obchodního tajemství

Odkazy

URL

Organizační jednotka

Přírodovědecká fakulta

Klíčová slova anglicky

bacteriophage jbd30, replication strategy, cryo-electron microscopy, tomography, fluorescent microscopy

Změněno: 12. 7. 2023 09:36, Ing. Lucie Valentová, Ph.D.

Anotace

V originále

Increasing numbers of infections caused by multi-drug resistant bacteria have renewed the interest in phage therapy. However, the exact mechanisms of phage – bacterium interaction are not known for the most of them. Here, we present the virion structure and infection strategy of Siphoviridae bacteriophage JBD30 revealed by the combination of cryo-electron and fluorescent microscopy. Bacteriophage JBD30 uses its tail fibres for recognising P. aeruginosa pili type IV. After the attachment to pili, the virion either diffuses along it or is pulled by pili retraction towards the cellular surface, where it irreversibly binds by the tripod complex of receptor binding protein trimers. Afterwards, the phage punctures the outer cellular membrane and degrades the peptidoglycan layer using the enzymatically active domains of receptor binding protein and tape measure protein C-terminal trimeric α-helical coiled coil domain. Bioinformatic analysis of tape measure protein showed that its N-terminal part is composed of three domains: hydrophobic transmembrane domain I (residues 57–79), cytoplasmic domain (residues 80–384) and hydrophobic transmembrane domain II (residues 385–409). Furthermore, the estimated size of these domains corresponds to the thickness of P. aeruginosa cellular membranes. We assume that the N-terminal part of the tape measure protein might form a channel spanning the whole cell wall facilitating DNA transition from the virion capsid into the host cytoplasm. New phage progeny is released approximately 80 minutes post ejection of phage DNA into the host cell. The combination of cryo-electron microscopy analysis techniques, cryo-electron tomography and fluorescent microscopy allowed us to propose the mechanism of key stages of phage infection and describe it at time resolved molecular level.

Návaznosti

LL1906, projekt VaV

Název: Replikace fágů v bakteriálním biofilmu

Investor: Ministerstvo školství, mládeže a tělovýchovy ČR, Replikace fágů v bakteriálním biofilmu

Citovat

VALENTOVÁ, Lucie, Tibor FÜZIK a Pavel PLEVKA. Carpe pili! Hunting strategy of phage JBD30 revealed by combination of cryo-electron and fluorescent microscopy. In XVIII Discussions in Structural Molecular Biology and 5th User Meeting of CIISB. 2023.

@proceedings{2297085,
   author = {Valentová, Lucie and Füzik, Tibor and Plevka, Pavel},
   booktitle = {XVIII Discussions in Structural Molecular Biology and 5th User Meeting of CIISB},
   keywords = {bacteriophage jbd30, replication strategy, cryo-electron microscopy, tomography, fluorescent microscopy},
   language = {eng},
   title = {Carpe pili! Hunting strategy of phage JBD30 revealed by combination of cryo-electron and fluorescent microscopy},
   url = {https://cssb.structbio.org/xix-discussions-in-structural-molecular-biology-and-6th-user-meeting-of-ciisb-czech-infrastructure-for-integrative-structural-biology/},
   year = {2023}
}

TY  - CONF
ID  - 2297085
AU  - Valentová, Lucie - Füzik, Tibor - Plevka, Pavel
PY  - 2023
TI  - Carpe pili! Hunting strategy of phage JBD30 revealed by combination of cryo-electron and fluorescent microscopy
KW  - bacteriophage jbd30, replication strategy, cryo-electron microscopy, tomography, fluorescent microscopy
UR  - https://cssb.structbio.org/xix-discussions-in-structural-molecular-biology-and-6th-user-meeting-of-ciisb-czech-infrastructure-for-integrative-structural-biology/
N2  - Increasing numbers of infections caused by multi-drug resistant bacteria have renewed the interest in phage therapy. However, the exact mechanisms of phage – bacterium interaction are not known for the most of them. Here, we present the virion structure and infection strategy of Siphoviridae bacteriophage JBD30 revealed by the combination of cryo-electron and fluorescent microscopy. Bacteriophage JBD30 uses its tail fibres for recognising P. aeruginosa pili type IV. After the attachment to pili, the virion either diffuses along it or is pulled by pili retraction towards the cellular surface, where it irreversibly binds by the tripod complex of receptor binding protein trimers. Afterwards, the phage punctures the outer cellular membrane and degrades the peptidoglycan layer using the enzymatically active domains of receptor binding protein and tape measure protein C-terminal trimeric α-helical coiled coil domain. Bioinformatic analysis of tape measure protein showed that its N-terminal part is composed of three domains: hydrophobic transmembrane domain I (residues 57–79), cytoplasmic domain (residues 80–384) and hydrophobic transmembrane domain II (residues 385–409). Furthermore, the estimated size of these domains corresponds to the thickness of P. aeruginosa cellular membranes. We assume that the N-terminal part of the tape measure protein might form a channel spanning the whole cell wall facilitating DNA transition from the virion capsid into the host cytoplasm. New phage progeny is released approximately 80 minutes post ejection of phage DNA into the host cell. The combination of cryo-electron microscopy analysis techniques, cryo-electron tomography and fluorescent microscopy allowed us to propose the mechanism of key stages of phage infection and describe it at time resolved molecular level.
ER  -

VALENTOVÁ, Lucie, Tibor FÜZIK a Pavel PLEVKA. Carpe pili! Hunting strategy of phage JBD30 revealed by combination of cryo-electron and fluorescent microscopy. In \textit{XVIII Discussions in Structural Molecular Biology and 5th User Meeting of CIISB}. 2023.

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