Direct interaction between fd phage pilot protein pIII and the TolQ–TolR proton-dependent motor provides new insights into the import of filamentous phages

Pellegri, Callypso; Moreau, Alain; Duché, Denis; Houot, Laetitia

doi:10.1016/j.jbc.2023.105048

Cited by 4 publications

(2 citation statements)

References 59 publications

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“…Of note, the F‐pilus as encoded on the F‐plasmid can promote DNA transfer in two directions: first, export of its single‐stranded DNA copy from the cytoplasm during horizontal gene transfer to recipient cells and, second, import of DNA via attachment of filamentous phage from outside. In contrast to the ATP‐driven DNA export during bacterial conjugation, a filamentous phage just utilizes the pilus tip as primary attachment site, which is followed by pilus retraction [ 85 , 86 ], whereas the pore‐forming and proton‐dependent tol system of the host cell then serves for the import of its circular single‐stranded DNA genome [ 87 ]. This explains why the DNA transfer system of conjugative plasmids, here affected by the traD36 mutation, is unimportant for the infection process of filamentous phage such as M13—which constitutes a crucial step in bacterial phage display technology.…”

Section: Resultsmentioning

confidence: 99%

Comparative genome analysis of three classical E. coli cloning strains designed for blue/white selection: JM83, JM109 and XL1‐Blue

Achatz,

Skerra

2024

FEBS Open Bio

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The development of the Escherichia coli K‐12 laboratory strains JM83, JM109 and XL1‐Blue was instrumental in early gene technology. We report the comprehensive genome sequence analysis of JM83 and XL1‐Blue using Illumina and Oxford Nanopore technologies and a comparison with both the wild‐type sequence (MG1655) and the genome of JM109 deposited at GenBank. Our investigation provides insight into the way how the genomic background that allows blue/white colony selection—by complementing a functionally inactive ω‐fragment of β‐galactosidase (LacZ) with its α‐peptide encoded on the cloning vector—has been implemented independently in these three strains using classical bacterial genetics. In fact, their comparative analysis reveals recurrent motifs: (i) inactivation of the native enzyme via large deletions of chromosomal regions encompassing the lac locus, or a chemically induced frameshift deletion at the beginning of the lacZ cistron, and (ii) utilization of a defective prophage (ϕ80), or an F′‐plasmid, to provide the lacZ∆M15 allele encoding its ω‐fragment. While the genetic manipulations of the E. coli strains involved repeated use of mobile genetic elements as well as harsh chemical or physical mutagenesis, the individual modified traits appear remarkably stable as they can be found even in distantly related laboratory strains, beyond those investigated here. Our detailed characterization at the genome sequence level not only offers clues about the mechanisms of classical gene transduction and transposition but should also guide the future fine‐tuning of E. coli strains for gene cloning and protein expression, including phage display techniques, utilizing advanced tools for site‐specific genome engineering.

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Section: Resultsmentioning

confidence: 99%

Comparative genome analysis of three classical E. coli cloning strains designed for blue/white selection: JM83, JM109 and XL1‐Blue

Achatz,

Skerra

2024

FEBS Open Bio

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“…The fd phage , a nanowire-like filamentous phage , infects bacteria specifically, posing no harm to humans (Pellegri et al 2023 ), featuring a diameter of approximately 6.5 nm and a length extending to several micrometers. The virus consists of approximately 2700 copies of a major coating protein, pVIII, distributed along the virus’s length, and five copies each of four minor coating proteins (pIII, pVI, pVII, and pIX) that cap the ends of the virus (Rakonjac et al 2023 , 2017 ).…”

Section: Introductionmentioning

confidence: 99%

Advanced detection of cervical cancer biomarkers using engineered filamentous phage nanofibers

Zhou,

Wang,

Bao

et al. 2024

Appl Microbiol Biotechnol

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Cervical cancer is a major global health concern, characterized by its high incidence and mortality rates. The detection of tumor markers is crucial for managing cancer, making treatment decisions, and monitoring disease progression. Vascular endothelial growth factor (VEGF) and programmed death-ligand 1 (PDL-1) are key targets in cervical cancer therapy and valuable biomarkers in predicting treatment response and prognosis. In this study, we found that combining the measurement of VEGF and soluble PDL-1 can be used for diagnosing and evaluating the progression of cervical cancer. To explore a more convenient approach for detecting and assessing cervical cancer, we designed and prepared an engineered fd bacteriophage, a human-safe viral nanofiber, equipped with two peptides targeting VEGF and PD-L1. The dual-display phage nanofiber specifically recognizes and binds to both proteins. Utilizing this nanofiber as a novel capture agent, we developed a new enzyme-linked immunosorbent assay (ELISA) method. This method shows significantly enhanced detection sensitivity compared to conventional ELISA methods, which use either anti-VEGF or anti-PD-L1 antibodies as capture agents. Therefore, the phage dual-display nanofiber presents significant potential in detecting cancer markers, evaluating medication efficacy, and advancing immunotherapy drug development. Key points • The combined measurement of VEGF and soluble Programmed Death-Ligand 1(sPD-L1) demonstrates an additive effect in the diagnosis of cervical cancer. Fd phage nanofibers have been ingeniously engineered to display peptides that bind to VEGF and PD-L1, enabling the simultaneous detection of both proteins within a single assay • Genetically engineered phage nanofibers, adorned with two distinct peptides, can be utilized for the diagnosis and prognosis of cancer and can be mass-produced cost-effectively through bacterial infections • Employing dual-display fd phage nanofibers as capture probes, the phage ELISA method exhibited significantly enhanced detection sensitivity compared to traditional sandwich ELISA. Furthermore, phage ELISA facilitates the detection of a single protein or the simultaneous detection of multiple proteins, rendering them powerful tools for protein analysis and diagnosis across various fields, including cancer research Graphical Abstract

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Protein–Protein Interactions: Oxidative Bacterial Two Hybrid

Pellegri,

Bouveret,

Houot

2023

Methods in Molecular Biology

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Direct interaction between fd phage pilot protein pIII and the TolQ–TolR proton-dependent motor provides new insights into the import of filamentous phages

Cited by 4 publications

References 59 publications

Comparative genome analysis of three classical E. coli cloning strains designed for blue/white selection: JM83, JM109 and XL1‐Blue

Comparative genome analysis of three classical E. coli cloning strains designed for blue/white selection: JM83, JM109 and XL1‐Blue

Advanced detection of cervical cancer biomarkers using engineered filamentous phage nanofibers

Protein–Protein Interactions: Oxidative Bacterial Two Hybrid

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