Selective phage infection mediated by epitope expression on F pilus 1 1Edited by J. Karn

Malmborg, Ann-Christin; Söderlind, Eskil; Frost, Laura S.; Borrebaeck, Carl

doi:10.1006/jmbi.1997.1332

Cited by 37 publications

(12 citation statements)

References 24 publications

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“…Therefore, one way to reduce the rate of mutation would be to deliver multiple killing systems, either by duplication of one system or by combining several different systems in the phagemid. In addition, since alteration or loss of the phage receptor (21) may also lead to resistance, it may be prudent to employ phage cocktails containing several different phages or a single genetically engineered phage that recognizes different cell surface receptors (22).…”

Section: Vol 47 2003 Lethal-agent Delivery System For Bacterial Infmentioning

confidence: 99%

Use of Genetically Engineered Phage To Deliver Antimicrobial Agents to Bacteria: an Alternative Therapy for Treatment of Bacterial Infections

Westwater

Kasman

Schofield

et al. 2003

Antimicrob Agents Chemother

199

123

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The emergence and increasing prevalence of multidrug-resistant bacterial pathogens emphasizes the need for new and innovative antimicrobial strategies. Lytic phages, which kill their host following amplification and release of progeny phage into the environment, may offer an alternative strategy for combating bacterial infections. In this study, however, we describe the use of a nonlytic phage to specifically target and deliver DNA encoding bactericidal proteins to bacteria. To test the concept of using phage as a lethal-agent delivery vehicle, we used the M13 phagemid system and the addiction toxins Gef and ChpBK. Phage delivery of lethal-agent phagemids reduced target bacterial numbers by several orders of magnitude in vitro and in a bacteremic mouse model of infection. Given the powerful genetic engineering tools available and the present knowledge in phage biology, this technology may have potential use in antimicrobial therapies and DNA vaccine development.

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Section: Vol 47 2003 Lethal-agent Delivery System For Bacterial Infmentioning

confidence: 99%

Use of Genetically Engineered Phage To Deliver Antimicrobial Agents to Bacteria: an Alternative Therapy for Treatment of Bacterial Infections

Westwater

Kasman

Schofield

et al. 2003

Antimicrob Agents Chemother

199

123

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“…The engineered g3p mutants were constructed by using the Chl r vector pAM18 (pACYC184 derived) and the g3p gene sequence from bacteriophage R408 (18,28). The expression from the pAM18 vector is under the control of a lac promoter.…”

Section: Bacterialmentioning

confidence: 99%

The Phage Infection Process: a Functional Role for the Distal Linker Region of Bacteriophage Protein 3

Nilsson

Malmborg

Borrebaeck

2000

J Virol

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The filamentous bacteriophage infects Escherichia coli by interaction with the F pilus and the TolQRA complex. The virus-encoded protein initiating this process is the gene 3 protein (g3p). The g3p molecule can be divided into three different domains separated by two glycine-rich linker regions. Though there has been extensive evaluation of the importance of the diverse domains of g3p, no proper function has so far been assigned to these linker regions. Through the design of mutated variants of g3p that were displayed on the surface of bacteriophage, we were able to elucidate a possible role for the distal glycine-rich linker region. A phage that displayed a g3p comprised of only the N1 domain, the first linker region, and the C-terminal domain was able to infect cells at almost the same frequency as the wild-type phage. This infection was proven to be dependent on the motif between amino acid residues 68 and 86 (i.e., the first glycine-rich linker region of g3p) and on F-pilus expression.Filamentous bacteriophage is a nonlytic DNA virus that infects Escherichia coli cells harboring an F episome (19,20,26). In order for bacteriophage infection to occur, including the translocation of the single-stranded phage DNA over the cell membrane, the expression of two different receptor systems in the gram-negative host cell is critical. The primary receptor, i.e., the F pilus, mediates the adsorption of the phage to the bacterial cell (6,12). This interaction can take place at a long distance from the cell surface, since the pilus molecule extrudes away from the bacteria. Through the depolymerization of the pilus, the bacteriophage is brought close to the cell surface of the host, where the interaction with the coreceptor takes place (5, 31). The coreceptor, i.e., the TolQRA complex, is localized in the inner membrane and the periplasmic space and is attached to the inside of the outer membrane (9, 16). It was recently shown that it is the C-terminal part of the TolA domain that interacts with the bacteriophage adsorption protein (7,27). Upon binding to the TolA domain, the viral DNA translocates into the host cell by an as-yet-unknown mechanism.Expression of the gene 3 viral adsorption protein, g3p, on the surface of the bacteriophage is essential for normal infection to occur (1). The minor coat protein g3p is located together with g6p in three to five copies at one end of the filamentous phage (10). Mature g3p consists of 406 amino acid residues separated in several distinct regions (2). The N-terminal part can be divided into two different domains, N1 and N2, mediating penetration and adsorption during infection, respectively (1, 30). The C-terminal part is responsible for the interaction with g6p, thereby anchoring the g3p in the membrane of the phage coat (1, 14, 21). Furthermore, the g3p has a crucial role in the assembly of the filamentous phage, since in the absence of g3p, the proper termination of the assembly and the following release of the phage will not take place (24).Two glycine-rich linker regions divid...

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“…It has also been shown to achieve specific enrichment factors of 10 10 after only two rounds of selection (Dueñas and Borrebaeck, 1994). The principle of using specific proteinligand interactions that result in a biologically measurable event (Malmborg et al, 1997) has great potential in, for example, functional genomics, where there is a need to rapidly characterize novel gene products or for in vitro evolution where very high binding affinities are desired. However, to use selective infection as a tool in these applications we need to understand what effect the affinity of the interacting pairs has on the process.…”

Section: Discussionmentioning

confidence: 99%

Selective infection of E. coli as a function of a specific molecular interaction

Nilsson

Karlsson

Rakonjac

et al. 2002

J of Molecular Recognition

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Selective infection of phage is when the bacterial infection depends on the specific molecular interaction between an antigen and a phage-displayed protein sequence such as an antibody. Engineering of the normal infection into pathways, directed by a specific protein--protein interaction, has raised several mechanistic questions. Here, we address the type of display and the affinity between the interacting pairs. The deleted phage R408d3 was used for the first time in selective infection and was shown to exhibit a superior performance compared to the VCSM13 phage. Furthermore, the affinity between the interacting pairs also affected the selective infection process and a correlation between affinity and infection efficiency was detected, thus implying that selective infection is the method of choice for selection of rare high-affinity interactions in molecular libraries.

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Selective phage infection mediated by epitope expression on F pilus 1 1Edited by J. Karn

Cited by 37 publications

References 24 publications

Use of Genetically Engineered Phage To Deliver Antimicrobial Agents to Bacteria: an Alternative Therapy for Treatment of Bacterial Infections

Use of Genetically Engineered Phage To Deliver Antimicrobial Agents to Bacteria: an Alternative Therapy for Treatment of Bacterial Infections

The Phage Infection Process: a Functional Role for the Distal Linker Region of Bacteriophage Protein 3

Selective infection of E. coli as a function of a specific molecular interaction

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