The molecular structure and structural transition of the α-helical capsid in filamentous bacteriophage Pf1

Welsh, Liam; Symmons, Martyn F.; Marvin, D.A.

doi:10.1107/s0907444999015334

Cited by 38 publications

(58 citation statements)

References 42 publications

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“…Structural fitting 21 of assigned experimental NMR data yields a unique three-dimensional structure of the protein. For the Pf1 coat protein structure described here (PDB entry 1PJF), the effective resolution determined by taking into account all sources of experimental error and uncertainties in the spin interaction tensors is estimated to be 1.5 Å , which compares favorably to the values reported for the structures determined by X-ray 5 fiber and neutron 7,8 diffraction. At atomic resolution, the structure of Pf1 coat protein is remarkably complex.…”

supporting

confidence: 56%

Structure of the Coat Protein in Pf1 Bacteriophage Determined by Solid-state NMR Spectroscopy

Thiriot

Nevzorov

Zagyanskiy

et al. 2004

Journal of Molecular Biology

134

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The atomic resolution structure of Pf1 coat protein determined by solidstate NMR spectroscopy of magnetically aligned filamentous bacteriophage particles in solution is compared to the structures previously determined by X-ray fiber and neutron diffraction, the structure of its membrane-bound form, and the structure of fd coat protein. These structural comparisons provide insights into several biological properties, differences between class I and class II filamentous bacteriophages, and the assembly process. The six N-terminal amino acid residues adopt an unusual "double hook" conformation on the outside of the bacteriophage particle. The solid-state NMR results indicate that at 30 8C, some of the coat protein subunits assume a single, fully structured conformation, and some have a few mobile residues that provide a break between two helical segments, in agreement with structural models from X-ray fiber and neutron diffraction, respectively. The atomic resolution structure determined by solid-state NMR for residues 7 -14 and 18 -46, which excludes the N-terminal double hook and the break between the helical segments, but encompasses more than 80% of the backbone including the distinct kink at residue 29, agrees with that determined by X-ray fiber diffraction with an RMSD value of 2.0 Å . The symmetry and distance constraints determined by X-ray fiber and neutron diffraction enable the construction of an accurate model of the bacteriophage particle from the coordinates of the coat protein monomers.

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supporting

confidence: 56%

Structure of the Coat Protein in Pf1 Bacteriophage Determined by Solid-state NMR Spectroscopy

Thiriot

Nevzorov

Zagyanskiy

et al. 2004

Journal of Molecular Biology

134

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“…Anti-Pf4 polyclonal antibodies were developed by using a synthetic peptide (Auspep, Parkville, Australia) with the following amino acid sequence: Gly-Val-Ile-Asp-Thr-Ser-Ala-Val-GluSer-Ala-Ile-Thr-Asp-Gly-Cys. This sequence corresponds to residues 1 to 15 of CoaB (PA0723; the major coat protein of the filamentous phage virion) of Pf1 (and of Pf4), which is exposed on the outer surface of the bacteriophage virion (35,61). An extra cysteine residue was added to the N terminus of this peptide for coupling to the carrier protein, keyhole limpet hemocyanin.…”

Section: Methodsmentioning

confidence: 99%

Bacteriophage and Phenotypic Variation in Pseudomonas aeruginosa Biofilm Development

2004

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A current question in biofilm research is whether biofilm-specific genetic processes can lead to differentiation in physiology and function among biofilm cells. In Pseudomonas aeruginosa, phenotypic variants which exhibit a small-colony phenotype on agar media and a markedly accelerated pattern of biofilm development compared to that of the parental strain are often isolated from biofilms. We grew P. aeruginosa biofilms in glass flow cell reactors and observed that the emergence of small-colony variants (SCVs) in the effluent runoff from the biofilms correlated with the emergence of plaque-forming Pf1-like filamentous phage (designated Pf4) from the biofilm. Because several recent studies have shown that bacteriophage genes are among the most highly upregulated groups of genes during biofilm development, we investigated whether Pf4 plays a role in SCV formation during P. aeruginosa biofilm development. We carried out immunoelectron microscopy using antiPf4 antibodies and observed that SCV cells, but not parental-type cells, exhibited high densities of Pf4 filaments on the cell surface and that these filaments were often tightly interwoven into complex latticeworks surrounding the cells. Moreover, infection of P. aeruginosa planktonic cultures with Pf4 caused the emergence of SCVs within the culture. These SCVs exhibited enhanced attachment, accelerated biofilm development, and large regions of dead and lysed cells inside microcolonies in a manner identical to that of SCVs obtained from biofilms. We concluded that Pf4 can mediate phenotypic variation in P. aeruginosa biofilms. We also performed partial sequencing and analysis of the Pf4 replicative form and identified a number of open reading frames not previously recognized in the genome of P. aeruginosa, including a putative postsegregational killing operon.

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“…Based on the calorimetry data, Hinz et al (18) hypothesized that one or more hydrophobic side chains change positions to lower their surface contact with water. Welsh et al (11), to account for their proposed conversion of a monomeric asymmetric unit in Pf1 L to a trimeric asymmetric unit in Pf1 H , suggested more extensive changes in subunit contacts and structure on the basis of proposed similarities to molecular crystals.…”

Section: Locations Of Atoms With Perturbed Chemical Shifts On Hydrophmentioning

confidence: 99%

“…One difference between Pf1 and the others that might account for the presence of a transition in Pf1 and not the others is its 1:1 stoichiometry. Diffraction studies of oriented phage in solution showed that the low and high temperature forms differ slightly in the orientation of the subunit with respect to the filament axis (10,11). The two states have slightly different numbers of subunits per turn in the helical arrangement of their capsids, which produce very different diffraction patterns even though the structures are closely similar.…”

mentioning

confidence: 99%

Intersubunit Hydrophobic Interactions in Pf1 Filamentous Phage

Goldbourt

Day²,

McDermott

2010

Journal of Biological Chemistry

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Magic angle spinning solid-state NMR has been used to study the structural changes in the Pf1 filamentous bacteriophage, which occur near 10°C. Comparisons of NMR spectra recorded above and below 10°C reveal reversible perturbations in many NMR chemical shifts, most of which are assigned to atoms of hydrophobic side chains of the 46-residue subunit. The changes mainly involve groups located in patches on the interfaces between neighboring capsid subunits. The observations show that the transition adjusts the hydrophobic interfaces between fairly rigid subunits. The low temperature form has been generally more amenable to structure determination; spin diffusion experiments on this form revealed unambiguous contacts between side chains of neighboring subunits. These contacts are important constraints for structure modeling.

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The molecular structure and structural transition of the α-helical capsid in filamentous bacteriophage Pf1

Cited by 38 publications

References 42 publications

Structure of the Coat Protein in Pf1 Bacteriophage Determined by Solid-state NMR Spectroscopy

Structure of the Coat Protein in Pf1 Bacteriophage Determined by Solid-state NMR Spectroscopy

Bacteriophage and Phenotypic Variation in Pseudomonas aeruginosa Biofilm Development

Intersubunit Hydrophobic Interactions in Pf1 Filamentous Phage

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