Tale of two spikes in bacteriophage PRD1

Huiskonen, Juha T.; Manole, Violeta; Butcher, Sarah J.

doi:10.1073/pnas.0608625104

Cited by 40 publications

(44 citation statements)

References 53 publications

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“…After SDS treatment, some minor amounts of P2 appeared at~140 kDa in the second dimension, as well as significant amounts of P5 (Table 1, Figs 3 and S1). These results are in accordance with the model suggesting that P5 and P2 proteins interact with each other as separate spikes, and that P5 attaches to the virus particle via P31 (Huiskonen et al, 2007). Under most conditions, P16 and P31 appeared partly as monomers (Table 1, Figs S1-S3).…”

supporting

confidence: 79%

Probing protein interactions in the membrane-containing virus PRD1

Mattila

Oksanen

Bamford

2015

Journal of General Virology

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PRD1 is a Gram-negative bacteria infecting complex tailless icosahedral virus with an inner membrane. This type virus of the family Tectiviridae contains at least 18 structural protein species, of which several are membrane associated. Vertices of the PRD1 virion consist of complexes recognizing the host cell, except for one special vertex through which the genome is packaged. Despite extensive knowledge of the overall structure of the PRD1 virion and several individual proteins at the atomic level, the locations and interactions of various integral membrane proteins and membrane-associated proteins still remain a mystery. Here, we demonstrated that blue native PAGE can be used to probe protein-protein interactions in complex membrane-containing viruses. Using this technique and PRD1 as a model, we identified the known PRD1 multiprotein vertex structure composed of penton protein P31, spike protein P5, receptor-binding protein P2 and stabilizing protein P16 linking the vertex to the internal membrane. Our results also indicated that two transmembrane proteins, P7 and P14, involved in viral nucleic acid delivery, make a complex. In addition, we performed a zymogram analysis using mutant particles devoid of the special vertex that indicated that the lytic enzyme P15 of PRD1 was not part of the packaging vertex, thus contradicting previously published results. INTRODUCTIONAmong the prokaryotic viruses infecting either archaea or bacteria, close to half of the described virus morphotypes contain lipid as a structural component (reviewed by Atanasova et al., 2015). Membrane-containing viruses are the most characteristic virus types among the eukaryoteinfecting viruses. One type of virus is the icosahedral internal membrane-containing morphotype. This type of virus is found infecting organisms of all domains of life, exemplified by bacteriophages PRD1, PM2 and P23-77 (Abrescia et al., 2004, 2008;Jaatinen et al., 2008), archaeal viruses SH1, Sulfolobus turreted icosahedral virus and Haloarcula hispanica icosahedral virus 2 (Jäälinoja et al., 2008;Jaakkola et al., 2012;Veesler et al., 2013), and Paramecium bursaria chlorella virus 1 and mimivirus of eukaryotes (Xiao et al., 2005;Zhang et al., 2011).Bacteriophage PRD1 is one of the best-known internal membrane-containing icosahedral viruses. It infects a variety of Gram-negative bacteria harbouring incompatibility P, N or W group plasmids (Olsen et al., 1974). PRD1 is the type member of the family Tectiviridae (for review, see Grahn et al., 2006;. The linear dsDNA genome of PRD1 (~15 kb) has inverted terminal repeats and covalently linked terminal proteins at the 59 ends (Bamford & Mindich, 1984;Bamford et al., 1983Savilahti & Bamford, 1986). The internal membrane consists of both protein and phospholipids iñ 1 : 1 mass ratio (Davis et al., 1982). Eighteen out of the 31 genes encode structural proteins either associated with the membrane (half of the protein species) or with the capsid shell (Fig. 1). The major capsid protein P3 (43 kDa) is a trimer organized on the ps...

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confidence: 79%

Probing protein interactions in the membrane-containing virus PRD1

Mattila

Oksanen

Bamford

2015

Journal of General Virology

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“…There are several possible explanations for low occupancy including steric hindrance, owing to the large size of the integrin molecules, and nonphysiological binding conditions. Together, the conformational heterogeneity and the low occupancy effectively abolish much of the integrin density when averaging approaches are used, as has been observed for viral spikes and virus-antibody and virus-receptor complexes (6,37,52,77). We have previously used classification analysis and vertex reconstruction to sort out such variation, but this was precluded here because of the size of the integrin with respect to the capsid (14, 36, 37, 40).…”

Section: Discussionmentioning

confidence: 99%

Interaction of α _V β ₃ and α _V β ₆ Integrins with Human Parechovirus 1

Seitsonen

Susi

Heikkilä

et al. 2010

J Virol

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Human parechovirus (HPEV) infections are very common in early childhood and can be severe in neonates. It has been shown that integrins are important for cellular infectivity of HPEV1 through experiments using peptide blocking assays and function-blocking antibodies to ␣ V integrins. The interaction of HPEV1 with ␣ V integrins is presumably mediated by a C-terminal RGD motif in the capsid protein VP1. We characterized the binding of integrins ␣ V ␤ 3 and ␣ V ␤ 6 to HPEV1 by biochemical and structural studies. We showed that although HPEV1 bound efficiently to immobilized integrins, ␣ V ␤ 6 bound more efficiently than ␣ V ␤ 3 to immobilized HPEV1. Moreover, soluble ␣ V ␤ 6, but not ␣ V ␤ 3, blocked HPEV1 cellular infectivity, indicating that it is a high-affinity receptor for HPEV1. We also showed that HPEV1 binding to integrins in vitro could be partially blocked by RGD peptides. Using electron cryo-microscopy and image reconstruction, we showed that HPEV1 has the typical T‫1؍‬ (pseudo T‫)3؍‬ organization of a picornavirus. Complexes of HPEV1 and integrins indicated that both integrin footprints reside between the 5-fold and 3-fold symmetry axes. This result does not match the RGD position predicted from the coxsackievirus A9 X-ray structure but is consistent with the predicted location of this motif in the shorter C terminus found in HPEV1. This first structural characterization of a parechovirus indicates that the differences in receptor binding are due to the amino acid differences in the integrins rather than to significantly different viral footprints.Picornaviruses consist of a positive-sense, single-stranded infectious RNA genome of approximately 7.3 kb enclosed in a capsid composed of 60 copies of each of the three or four capsid proteins (VP1 to VP4). Human parechovirus 1 (HPEV1) is a member of the Parechovirus genus of the Picornaviridae family (38, 70). There are currently eight completely sequenced human parechovirus types and 14 described types (4,19,24,30,38,39,51,58,78). In addition, the Parechovirus genus currently has four Ljungan virus members that infect rodents. HPEV1 exhibits several distinct molecular characteristics compared to other picornaviruses (38, 71). These include the lack of the maturation cleavage of the capsid proteins VP0 to VP4 (Nterminal) and VP2 (C-terminal), existence of an approximately 30-amino-acid-long extension to the N terminus of VP3, a unique nonstructural protein 2A, and a 5Ј untranslated region that is more closely related to picornaviruses infecting animals than those infecting humans.HPEV infections are common during the first years of life and are often mild or asymptomatic (20,28,42,73,80). Recently, a number of new types have been identified, and their prevalence in stool samples, for example, highlights their clinical importance. Normally, they cause gastroenteritis and respiratory infections, but severe illnesses, such as infections of the central nervous system, generalized infections of neonates, and myocarditis, have also been associated with HPEV infect...

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“…Since C557 and A223 homologues are present in STIV2 (see Table S2 in the supplemental material) but C381 is found only in STIV, we propose that the STIV turret ears are most likely formed from C381. The turrets thus remind us of the T-even phage tail fibers and the spike protein, P5, of phage PRD1, which are important for increasing the efficiency of infection by helping in host cell recognition but are not essential for irreversible attachment to the host (21,22,61).…”

Section: Discussionmentioning

confidence: 99%

Familial Relationships in Hyperthermo- and Acidophilic Archaeal Viruses

Happonen

Redder

Peng

et al. 2010

J Virol

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Archaea often live in extreme, harsh environments such as acidic hot springs and hypersaline waters. To date, only two icosahedrally symmetric, membrane-containing archaeal viruses, SH1 and Sulfolobus turreted icosahedral virus (STIV), have been described in detail. We report the sequence and three-dimensional structure of a third such virus isolated from a hyperthermoacidophilic crenarchaeon, Sulfolobus strain G4ST-2. Characterization of this new isolate revealed it to be similar to STIV on the levels of genome and structural organization. The genome organization indicates that these two viruses have diverged from a common ancestor. Interestingly, the prominent surface turrets of the two viruses are strikingly different. By sequencing and mass spectrometry, we mapped several large insertions and deletions in the known structural proteins that could account for these differences and showed that both viruses can infect the same host. A combination of genomic and proteomic analyses revealed important new insights into the structural organization of these viruses and added to our limited knowledge of archaeal virus life cycles and host-cell interactions.

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Tale of two spikes in bacteriophage PRD1

Cited by 40 publications

References 53 publications

Probing protein interactions in the membrane-containing virus PRD1

Probing protein interactions in the membrane-containing virus PRD1

Interaction of α _V β ₃ and α _V β ₆ Integrins with Human Parechovirus 1

Familial Relationships in Hyperthermo- and Acidophilic Archaeal Viruses

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Tale of two spikes in bacteriophage PRD1

Cited by 40 publications

References 53 publications

Probing protein interactions in the membrane-containing virus PRD1

Probing protein interactions in the membrane-containing virus PRD1

Interaction of α V β 3 and α V β 6 Integrins with Human Parechovirus 1

Familial Relationships in Hyperthermo- and Acidophilic Archaeal Viruses

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Interaction of α _V β ₃ and α _V β ₆ Integrins with Human Parechovirus 1