Crystal Structure of Bacteriophage SPP1 Distal Tail Protein (gp19.1)

Veesler, David; Robin, Gautier; Lichière, Julie; Auzat, Isabelle; Tavares, Paulo; Bron, Patrick; Campanacci, Valérie; Cambillau, Christian

doi:10.1074/jbc.m110.157529

Cited by 73 publications

(68 citation statements)

References 49 publications

Supporting

Mentioning

Contrasting

Order By: Relevance

“…Below the Dit density, a dome-like structure is observed, with its tip in distal position. Both volume and shape of this EM density are comparable with those observed in the equivalent position in the SPP1 virion EM reconstruction (5,31). The shape of this density is also reminiscent of the ORF16 one in phage p2 which is directly located under ORF15, a Dit homologue (17).…”

Section: Table 1 Correlation Coefficient (Cc) Values Resulting From Fsupporting

confidence: 58%

See 1 more Smart Citation

Structure and Molecular Assignment of Lactococcal Phage TP901-1 Baseplate

Bebeacua

Bron

Lai

et al. 2010

Journal of Biological Chemistry

Self Cite

View full text Add to dashboard Cite

P335 lactococcal phages infect the Gram؉ bacterium Lactococcus lactis using a large multiprotein complex located at the distal part of the tail and termed baseplate (BP). The BP harbors the receptor-binding proteins (RBPs), which allow the specific recognition of saccharidic receptors localized on the host cell surface. We report here the electron microscopic structure of the phage TP901-1 wild-type BP as well as those of two mutants bppL ؊ and bppU ؊ , lacking BppL (the RBPs) or both peripheral BP components (BppL and BppU), respectively. We also achieved an electron microscopic reconstruction of a partial BP complex, formed by BppU and BppL. This complex exhibits a tripod shape and is composed of nine BppLs and three BppUs. These structures, combined with lightscattering measurements, led us to propose that the TP901-1 BP harbors six tripods at its periphery, located around the central tube formed by ORF46 (Dit) hexamers, at its proximal end, and a ORF47 (Tal) trimer at its distal extremity. A total of 54 BppLs (18 RBPs) are thus available to mediate host anchoring with a large apparent avidity. TP901-1 BP exhibits an infection-ready conformation and differs strikingly from the lactococcal phage p2 BP, bearing only 6 RBPs, and which needs a conformational change to reach its activated state. The comparison of several Siphoviridae structures uncovers a close organization of their central BP core whereas striking differences occur at the periphery, leading to diverse mechanisms of host recognition.The first steps of phage infection require interactions between the phage receptor-binding proteins (RBPs) 2 (1, 2) and the receptors at the host cell surface. Although some RBPs are located at the tip of fibers (3), others belong to an elongated structure, the tail spike (4, 5). In bacteriophages infecting the Gram ϩ bacterium Lactococcus lactis such as p2 (936 group), TP901-1, and Tuc2009 (P335 group), RBPs are part of a large organelle (1-2 MDa) termed the baseplate (BP). We previously solved RBP structures of phages p2 (6, 7) and TP901-1 (8) as well as the RBP C-terminal domain ("head domain") of phage bIL170 (936 group) (9). It appeared that the RBP of phage TP901-1 (termed BppL, lower baseplate protein) was cleaved during crystallization, and the polypeptidic chain in the crystal structure starts either at residue 16 or at residue 32 (10). Proteolytic cleavage was also observed for the homologous RBP from Tuc2009 (11) as well as in the structure of a chimeric RBP comprising the N-terminal and linker domains of phage TP901-1 RBP fused to the C-terminal domain of phage p2 RBP (12). We demonstrated that individually expressed Tuc2009 BppU (upper baseplate protein) and BppL did not interact when mixed, and we attributed this to the proteolytic cleavage of the BppL N terminus that should normally plug into BppU (13). In contrast, co-expression of BppU and BppL yielded a well defined 3:9 complex (13).The BP architecture of the P335 phages TP901-1 and Tuc2009 has been investigated thoroughly using mutagenesis and immuno...

show abstract

Section: Table 1 Correlation Coefficient (Cc) Values Resulting From Fsupporting

confidence: 58%

“…S4). We previously determined the x-ray structure of the Dit protein (gp19.1) from phage SPP1 (31). In this structure, the N-terminal domains form two hexameric rings stacked back to back, whereas the C-terminal domains form a separate lectin-like domain located at the periphery of the rings.…”

Section: Table 1 Correlation Coefficient (Cc) Values Resulting From Fmentioning

confidence: 99%

Structure and Molecular Assignment of Lactococcal Phage TP901-1 Baseplate

Bebeacua

Bron

Lai

et al. 2010

Journal of Biological Chemistry

Self Cite

View full text Add to dashboard Cite

show abstract

“…Finally, a 34-Å-wide central channel is present in the center of the tail tube, aligned with the connector and baseplate channels, forming the dsDNA genome ejection pathway (17,(47)(48)(49)(50)(51) (Fig. 5G).…”

Section: Sequence Analysismentioning

confidence: 99%

Cryo-Electron Microscopy Structure of Lactococcal Siphophage 1358 Virion

Spinelli

Bebeacua

Orlov

et al. 2014

J Virol

Self Cite

View full text Add to dashboard Cite

Lactococcus lactis, a Gram؉ lactic acid-producing bacterium used for the manufacture of several fermented dairy products, is subject to infection by diverse virulent tailed phages, leading to industrial fermentation failures. This constant viral risk has led to a sustained interest in the study of their biology, diversity, and evolution. Lactococcal phages now constitute a wide ensemble of at least 10 distinct genotypes within the Caudovirales order, many of them belonging to the Siphoviridae family. Lactococcal siphophage 1358, currently the only member of its group, displays a noticeably high genomic similarity to some Listeria phages as well as a host range limited to a few L. lactis strains. These genomic and functional characteristics stimulated our interest in this phage. Here, we report the cryo-electron microscopy structure of the complete 1358 virion. Phage 1358 exhibits noteworthy features, such as a capsid with dextro handedness and protruding decorations on its capsid and tail. Observations of the baseplate of virion particles revealed at least two conformations, a closed and an open, activated form. Functional assays uncovered that the adsorption of phage 1358 to its host is Ca 2؉ independent, but this cation is necessary to complete its lytic cycle. Taken together, our results provide the complete structural picture of a unique lactococcal phage and expand our knowledge on the complex baseplate of phages of the Siphoviridae family. IMPORTANCE Phages of Lactococcus lactis are investigated mainly because they are sources of milk fermentation failures in the dairy industry.Despite the availability of several antiphage measures, new phages keep emerging in this ecosystem. In this study, we provide the cryo-electron microscopy reconstruction of a unique lactococcal phage that possesses genomic similarity to particular Listeria phages and has a host range restricted to only a minority of L. lactis strains. The capsid of phage 1358 displays the almost unique characteristic of being dextro handed. Its capsid and tail exhibit decorations that we assigned to nonspecific sugar binding modules. We observed the baseplate of 1358 in two conformations, a closed and an open form. We also found that the adsorption to its host, but not infection, is Ca 2؉ independent. Overall, this study advances our understanding of the adhesion mechanisms of siphophages.

show abstract

“…In siphophages, high-resolution structures of the host adsorption device of the Lactococcus phages p2 (5) and TP901-1 (6), and part of it for the Bacillus subtilis phage SPP1 (7,8), are available: they are made up of a complex baseplate containing multiple copies of the saccharide-binding RBP (18 and 54 for phages p2 and TP901-1, respectively) or a tail spike containing three copies of the protein-binding RBP (SPP1) (9). In these bacteriophages, which infect Gram-positive bacteria, a common docking hub between the tail tube and the tail adsorption device is the Dit-Tal complex (8).…”

mentioning

confidence: 99%

Crystal Structure of pb9, the Distal Tail Protein of Bacteriophage T5: a Conserved Structural Motif among All Siphophages

Flayhan

Vellieux

Lurz

et al. 2014

J Virol

View full text Add to dashboard Cite

fThe tail of Caudovirales bacteriophages serves as an adsorption device, a host cell wall-perforating machine, and a genome delivery pathway. In Siphoviridae, the assembly of the long and flexible tail is a highly cooperative and regulated process that is initiated from the proteins forming the distal tail tip complex. In Gram-positive-bacterium-infecting siphophages, the distal tail (Dit) protein has been structurally characterized and is proposed to represent a baseplate hub docking structure. It is organized as a hexameric ring that connects the tail tube and the adsorption device. In this study, we report the characterization of pb9, a tail tip protein of Escherichia coli bacteriophage T5. By immunolocalization, we show that pb9 is located in the upper part of the cone of the T5 tail tip, at the end of the tail tube. The crystal structure of pb9 reveals a two-domain protein. Domain A exhibits remarkable structural similarity with the N-terminal domain of known Dit proteins, while domain B adopts an oligosaccharide/ oligonucleotide-binding fold (OB-fold) that is not shared by these proteins. We thus propose that pb9 is the Dit protein of T5, making it the first Dit protein described for a Gram-negative-bacterium-infecting siphophage. Multiple sequence alignments suggest that pb9 is a paradigm for a large family of Dit proteins of siphophages infecting mostly Gram-negative hosts. The modular structure of the Dit protein maintains the basic building block that would be conserved among all siphophages, combining it with a more divergent domain that might serve specific host adhesion properties.

show abstract

Crystal Structure of Bacteriophage SPP1 Distal Tail Protein (gp19.1)

Cited by 73 publications

References 49 publications

Structure and Molecular Assignment of Lactococcal Phage TP901-1 Baseplate

Structure and Molecular Assignment of Lactococcal Phage TP901-1 Baseplate

Cryo-Electron Microscopy Structure of Lactococcal Siphophage 1358 Virion

Crystal Structure of pb9, the Distal Tail Protein of Bacteriophage T5: a Conserved Structural Motif among All Siphophages

Contact Info

Product

Resources

About