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Assembly of bacteriophage PRD1 spike complex: role of the multidomain protein P5
J Caldentey1, R Tuma, D H Bamford
1Institute of Biotechnology and Department of Biosciences, Viikki Biocenter, P.O. Box 56 (Viikinkaari 5), 00014 University of Helsinki, Finland. javier.caldentey@helsinki.fi
Abstract:
The spike structure of bacteriophage PRD1 is comprised of proteins P2, P5, and P31. It resembles the corresponding receptor-binding structure of adenoviruses. We show that purified recombinant protein P5 is an elongated (30 x 2.7 nm; R(h) = 5.5 nm), multidomain trimer which can slowly associate into nonamers. Cleavage of the 340 amino acid long P5 with collagenase yields 2 fragments. The larger, 205 amino acid long C-terminal fragment appears to contain the residues responsible for the trimerization of the protein, whereas the smaller N-terminal part mediates the interaction of P5 with the pentameric vertex protein P31 (24 x 2.5 nm, R(h) = 4.2 nm). In addition, the presence of the N-terminal sequence is required for the formation of the P5 nonamer. The results presented here suggest that P5 and P31 form an elongated adaptor complex at the 5-fold vertexes of the virion which anchors the adsorption protein P2 (21 x 2.5 nm; R(h) = 4.1 nm). Our results also suggest that the P5 trimer forms a substantial part of the viral spike shaft that was previously thought to be composed exclusively of protein P2.
Insights
Bacteriophage PRD1 spike proteins P5 and P31 form an adaptor complex. This complex anchors the adsorption protein P2 and suggests P5 forms part of the viral spike shaft.
Area of Science:
- Structural biology
- Virology
- Biochemistry
Background:
- The bacteriophage PRD1 spike structure shares similarities with adenovirus structures.
- Understanding the composition and assembly of viral spikes is crucial for virology.
Purpose of the Study:
- To elucidate the structural roles of proteins P5 and P31 in the bacteriophage PRD1 spike complex.
- To investigate the assembly and interactions of these proteins.
Main Methods:
- Purification and characterization of recombinant protein P5.
- Limited proteolysis using collagenase to identify functional domains.
- Analysis of protein interactions and complex formation.
Main Results:
- Protein P5 is an elongated trimer that can form nonamers, with its C-terminus mediating trimerization and N-terminus mediating interaction with P31.
- Protein P31 is a pentameric vertex protein.
- P5 and P31 form an elongated adaptor complex at the virion's 5-fold vertices, anchoring protein P2.
- The P5 trimer contributes significantly to the viral spike shaft structure.
Conclusions:
- Protein P5 and P31 form a key adaptor complex in the bacteriophage PRD1 spike.
- Protein P5 plays a dual role in spike assembly, contributing to both the shaft and vertex structure.
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