Filamentous phage structure, infection and assembly

Marvin, D.A.

doi:10.1016/s0959-440x(98)80032-8

Cited by 323 publications

(225 citation statements)

References 72 publications

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“…The criterion for a peak to be isolated is more stringent for dynamic assignments than for chemical assignments because spectral congestion in the dipolar dimension can easily pollute the accuracy of the dipolar fit. In this study nonoverlapping cross-peaks within a window of Ϯ0.25 ppm in both 13 C dimensions were accepted. For reference, a 2D 13 C-13 C spectrum from the Lee-Goldburg cross-polarization with dipolar assisted rotational resonance (LGCP-DARR) experiment is provided in Figs.…”

Section: Resultsmentioning

confidence: 99%

“…The virion consists of a circular single-stranded DNA genome of 7,349 nt (3) within a cylindrical capsid of Ϸ7,300 major coat protein subunits that have 1:1 stoichiometric interactions (4,5) with nucleotides along the filament, as well as a small number of other proteins interacting with reverse turns of DNA at the ends. A variety of spectroscopic and diffraction studies of Pf1 have been carried out (5)(6)(7)(8)(9)(10)(11)(12)(13)(14)(15)(16)(17)(18)(19), including studies by solid-state NMR of the protein in the intact virion (16)(17)(18)(19), as in the present study, and by solution NMR of the isolated protein in detergent micelles (8,14) as a model of its structure in the membrane before virion assembly. Virion assembly occurs at the membrane in such a way that major coat protein subunits are packed in the filament as ␣-helices with their Nterminal regions on the outside and their C-terminal regions buried in the interior to interact with the DNA.…”

mentioning

confidence: 99%

“…There are contradictory data and unresolved questions in both aspects of the protein structure. For example, it has been recently proposed that there is a minor ''outward-bulge'' conformer in the backbone of residues 15-17 that is highly dynamic (16), but such a minor conformer was not observed in the 13 C and 15 N chemical shift assignments (18). Is the backbone indeed dynamic in this region?…”

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confidence: 99%

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Conformational dynamics of an intact virus: Order parameters for the coat protein of Pf1 bacteriophage

Lorieau

Day²,

McDermott

2008

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

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This study has examined the atomic-level dynamics of the protein in the capsid of filamentous phage Pf1. This capsid consists of Ϸ7,300 small subunits of only 46 aa in a helical array around a highly extended, circular single-stranded DNA molecule of 7,349 nt. Measurements were made of site-specific, solid-state NMR order parameters, ͗S͘, the values which are dimensionless quantities between 0 (mobile) and 1 (static) that characterize the amplitudes of molecular bond angular motions that are faster than microseconds. It was found that the protein subunit backbone is very static, and of particular interest, it appears to be static at residues glycine 15 and glutamine 16 where it had been previously thought to be mobile. In contrast to the backbone, several side chains display large-amplitude angular motions. Side chains on the virion exterior that interact with solvent are highly mobile, but surprisingly, the side chains of residues arginine 44 and lysine 45 near the DNA deep in the interior of the virion are also highly mobile. The largeamplitude dynamic motion of these positively charged side chains in their interactions with the DNA were not previously expected. The results reveal a highly dynamic aspect of a DNA-protein interface within a virus.solid-state NMR ͉ motion ͉ side chain T he Pf1 bacteriophage is a 2-m-long filamentous virus that infects Pseudomonas aeruginosa strain K (1, 2). The virion consists of a circular single-stranded DNA genome of 7,349 nt (3) within a cylindrical capsid of Ϸ7,300 major coat protein subunits that have 1:1 stoichiometric interactions (4, 5) with nucleotides along the filament, as well as a small number of other proteins interacting with reverse turns of DNA at the ends. A variety of spectroscopic and diffraction studies of Pf1 have been carried out (5-19), including studies by solid-state NMR of the protein in the intact virion (16)(17)(18)(19), as in the present study, and by solution NMR of the isolated protein in detergent micelles (8,14) as a model of its structure in the membrane before virion assembly. Virion assembly occurs at the membrane in such a way that major coat protein subunits are packed in the filament as ␣-helices with their Nterminal regions on the outside and their C-terminal regions buried in the interior to interact with the DNA.One of the studies of the Pf1 virion by solid-state NMR was on magnetically aligned hydrated virions, and it characterized the path and orientation of the backbone of the ␣-helical major capsid protein subunit by means of 1 H-15 N polarization inversion spin exchange at the magic angle (PISEMA) (16,17). It was proposed that the subunit consists of an N terminus that forms a double hook, a C terminus with an unraveled ␣-helix, and a central portion of three ␣-helices with two bends near the center. An assumed capsid helical symmetry for the high-temperature form of the virion (6, 20) was used to generate the intersubunit contacts for a model of the capsid (16,17). In a more recent solid-state NMR study from our group (18, 19), m...

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Section: Resultsmentioning

confidence: 99%

mentioning

confidence: 99%

mentioning

confidence: 99%

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Conformational dynamics of an intact virus: Order parameters for the coat protein of Pf1 bacteriophage

Lorieau

Day²,

McDermott

2008

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

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“…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%

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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“…They consist of a single-stranded circular DNA packed in a tube composed of the major coat protein pVIII (98% by mass), and a few copies of the minor coat proteins capping the ends of the phage particle (reviewed in [8,9]). …”

Section: Phage As Bioselective Nanomaterialsmentioning

confidence: 99%

Landscape phage as a molecular recognition interface for detection devices

Petrenko

2008

Microelectronics Journal

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Filamentous phages are thread-shaped bacterial viruses. Their outer coat is a tube formed by thousands equal copies of the major coat protein pVIII. Libraries of random peptides fused to pVIII domains were used for selection of phages probes specific for a panel of test antigens and biological threat agents. Because the viral carrier in the phage borne bio-selective probes is infective, they can be cloned individually and propagated indefinitely without needs of their chemical synthesis or reconstructing. As a new bioselective material, landscape phages combine unique characteristics of affinity reagents and self assembling proteins. Biorecognition layers formed by the phage-derived probes bind biological agents with high affinity and specificity and generate detectable signals in analytical platforms. The performance of phage-derived materials as biorecognition interface was illustrated by detection of Bacillus anthracis spores and Salmonella typhimurium cells. With further refinement, the phage-derived analytical platforms for detecting and monitoring of numerous threat agents may be developed, since phage interface against any bacteria, virus or toxin may be readily selected from the landscape phage libraries. As an interface in the field-use detectors, they may be superior to antibodies, since they are inexpensive, highly specific and strong binders, resistant to high temperatures and environmental stresses.

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Filamentous phage structure, infection and assembly

Cited by 323 publications

References 72 publications

Conformational dynamics of an intact virus: Order parameters for the coat protein of Pf1 bacteriophage

Conformational dynamics of an intact virus: Order parameters for the coat protein of Pf1 bacteriophage

Characterization of MDAΦ, a temperate filamentous bacteriophage of Neisseria meningitidis

Landscape phage as a molecular recognition interface for detection devices

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