Lcoation of Filamentous Phage Minor Coat Proteins in Phage and in Infected Cells

Endemann, Heike; Model, Peter

doi:10.1006/jmbi.1995.0393

Cited by 107 publications

(90 citation statements)

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“…However, in the case of the orf3 mutant, detection of a low amount of secreted forms was unexpected as this gene encodes a protein with homologies to pVII, a minor coat protein of phage Ff that caps the first end of the phage and has been shown to be necessary for its packaging. However, Endemann and Model have demonstrated that the absence of pVII leads to a drastic reduction of phage Ff production (Endemann & Model, 1995), and to the secretion of low amounts of phages, which is consistent with our observations. Altogether, these data demonstrate that the genomic organization of the MDA island is very close to that of Ff phages.…”

Section: Characterization Of the Orfs Of The Mda Islandsupporting

confidence: 82%

“…Results were compared with the consequences of mutations in genes located at similar places in M13, a filamentous phage of the Ff family, and are reported in Table 5 (Marvin & Hohn, 1969). (Endemann & Model, 1995;Lopez & Webster, 1983;Marvin, 1998). DDetection of the ds cytosolic form was performed by PCR using oligonucleotides 6-F with EXT1-b and EXT9-b with 6-R. §Detection of the ss supernatant form was performed by PCR using oligonucleotides 6-F with EXT1-b and EXT9-b with 6-R.…”

Section: Characterization Of the Orfs Of The Mda Islandmentioning

confidence: 99%

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Characterization of MDAΦ, a temperate filamentous bacteriophage of Neisseria meningitidis

et al. 2016

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The mechanism by which Neisseria meningitidis becomes invasive is not well understood. Comparative genomics identified the presence of an 8 kb island in strains belonging to invasive clonal complexes. This island was designated MDA for meningococcal disease associated. MDA is highly conserved among meningococcal isolates and its analysis revealed a genomic organization similar to that of a filamentous prophage such as CTXW of Vibrio cholerae. Subsequent molecular investigations showed that the MDA island has indeed the characteristics of a filamentous prophage, which can enter into a productive cycle and is secreted using the type IV pilus (tfp) secretin PilQ. At least three genes of the prophage are necessary for the formation of the replicative cytoplasmic form (orf1, orf2 and orf9). Immunolabelling of the phage with antibodies against the major capsid protein, ORF4, confirmed that filamentous particles, about 1200 nm long, covered with ORF4 are present at the bacterial surface forming bundles in some places and interacting with pili. The MDA bacteriophage is able to infect different N. meningitidis strains, using the type IV pili as a receptor via an interaction with the adsorption protein ORF6. Altogether, these data demonstrate that the MDA island encodes a functional prophage able to produce infectious filamentous phage particles.

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Section: Characterization Of the Orfs Of The Mda Islandsupporting

confidence: 82%

Section: Characterization Of the Orfs Of The Mda Islandmentioning

confidence: 99%

Characterization of MDAΦ, a temperate filamentous bacteriophage of Neisseria meningitidis

et al. 2016

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“…Then, of greater significance, we showed that antibody variable regions fused to pVII and pIX engaged in a dynamic interaction to display a functional Fv antibody, a representative heterodimeric motif. The pIII, pVI, pVIII, pVII, and pIX coat proteins are integral inner membrane proteins before assembly (24). With the advent of our results, all five now have been used for displaying proteins on the phage surface.…”

Section: Discussionmentioning

confidence: 99%

Making artificial antibodies: A format for phage display of combinatorial heterodimeric arrays

Gao

Mao

et al. 1999

Proc. Natl. Acad. Sci. U.S.A.

151

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The gene VII protein (pVII) and gene IX protein (pIX) are associated closely on the surface of filamentous bacteriophage that is opposite of the end harboring the widely exploited pIII protein. We developed a phagemid format wherein antibody heavy-and light-chain variable regions were fused to the amino termini of pVII and pIX, respectively. Significantly, the fusion proteins interacted to form a functional Fv-binding domain on the phage surface. Our approach will be applicable to the display of generic peptide and protein libraries that can form combinatorial heterodimeric arrays. Consequently, it represents a first step toward artificial antibodies and the selection of novel biological activities.Now that combinatorial antibody libraries have been secured (1-9), a powerful next step is the evolution toward artificial antibody constructs. Artificial antibodies are defined here as protein motifs of large diversity that use the functional strategy of the antibody molecule, but can be free of loop and framework structural constraints. When reduced to its essence, the antibody molecule is a biological device for the presentation of a combinatorial array of peptide elements in threedimensional space. The essential feature is that while CDRs (complementarity-determining regions) cooperate to form a binding site, their interaction is dynamic and functional with little structural association between the CDRs themselves. In this way, the full complement of amino acid residues is available for antigen recognition at a minimum energetic cost for binding. We propose that the ability to control the combinatorial design not only of sequence space but also of three-dimensional space would recapitulate and ultimately transcend the natural design of the immune repertoire.Although phage display has been investigated intensively, many details of the phage particle itself have not been fully elucidated, and the possibility of alternative display formats also remain to be explored. The filamentous bacteriophage fd, and, similarly, M13, consists of a circular, single-stranded DNA molecule surrounded by a cylinder of coat proteins (Fig. 1). The molecular mass of a particle is about 1.6 ϫ 10 7 Da, of which 88% is protein and 12% is DNA (10). There are about 2,700 molecules of the major coat protein pVIII that envelope the phage. At one end of the particle, there are five copies each of gene VII and VI proteins (pIII and pVI) that are involved in host-cell binding and in the termination of the assembly process. The other end contains five copies each of pVII and pIX that are actually hydrophobic peptides of 33 and 32 aa, respectively, required for the initiation of assembly and for maintenance of virion stability. Whereas pIII, pVI, and pVIII have been used to display biological molecules, pVII and pIX have not been utilized (1, 11).Substantial information has been accumulated about the structural and the phage display characteristics of pIII and pVIII. Yet, the data concerning the minor proteins encoded by genes VII and IX are...

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“…Unlike most bacterial viruses, which are assembled in the cytoplasm and released by cell lysis, filamentous phage are coordinately exported as they are assembled at the cell surface in a secretory process that leaves the cells fully viable (1). The five structural proteins of the virus are anchored in the inner membrane before their incorporation into the phage particle (2), and the phage genome is present in the cytoplasm. As assembly begins, the phage genome traverses the inner and outer membranes, picking up two minor coat proteins, pVII and pIX, at the leading tip (3) and a helical array of major coat protein subunits (pVIII) along its length.…”

mentioning

confidence: 99%

Assembling filamentous phage occlude pIV channels

Marciano

Russel

Simon

2001

Proc. Natl. Acad. Sci. U.S.A.

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Filamentous phage f1 is exported from its Escherichia coli host without killing the bacterial cell. Phage-encoded protein pIV, which is required for phage assembly and secretion, forms large highly conductive channels in the outer membrane of E. coli. It has been proposed that the phage are extruded across the bacterial outer membrane through pIV channels. To test this prediction, we developed an in vivo assay by using a mutant pIV that functions in phage export but whose channel opens in the absence of phage extrusion. In E. coli lacking its native maltooligosacharride transporter LamB, this pIV variant allowed oligosaccharide transport across the outer membrane. This entry of oligosaccharide was decreased by phage production and still further decreased by production of phage that cannot be released from the cell surface. Thus, exiting phage block the pIV-dependent entry of oligosaccharide, suggesting that phage occupy the lumen of pIV channels. This study provides the first evidence, to our knowledge, for viral exit through a large aqueous channel.

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Lcoation of Filamentous Phage Minor Coat Proteins in Phage and in Infected Cells

Cited by 107 publications

References 0 publications

Characterization of MDAΦ, a temperate filamentous bacteriophage of Neisseria meningitidis

Characterization of MDAΦ, a temperate filamentous bacteriophage of Neisseria meningitidis

Making artificial antibodies: A format for phage display of combinatorial heterodimeric arrays

Assembling filamentous phage occlude pIV channels

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