Abstract—
A modified method for culturing, concentrating, and purifying phage ϕ24B preparations was developed. In particular, a new lysogenic phage-producing strain lacking flagella was used, induction conditions were optimized, and purification in a sucrose gradient followed by concentration by deposition on a Freon 113 cushion were used. Using this method, a preparation of the Stx-converting bacteriophage ϕ24B was obtained, suitable for CryoEM direct analysis. Based on CryoEM data for this phage, the first primary three-dimensional reconstruction of its virions was performed. The structure of the phage ϕ24B tail is described. It was shown that the adsorption apparatus of this virus is represented by six thin lateral fibrils, and an axial fibril located at the end of the tail. This arrangement of the tail structure is consistent with the previously proposed hypothesis based on analysis of the receptor binding proteins (RBPs) of this bacteriophage.
This is a preview of subscription content, log in via an institution to check access.
Similar content being viewed by others
REFERENCES
Allison, H.E., Sergeant, M.J., James, C.E., Saunders, J.R., Smith, D.L., Sharp, R.J., Marks, T.S., and McCarthy, A.J., Immunity profiles of wild-type and recombinant shiga-like toxin-encoding bacteriophages and characterization of novel double lysogens, Infect. Immun., 2003, vol. 71, no. 6, pp. 3409‒3418.
Blake, K.S., Choi, J., and Dantas, G., Approaches for characterizing and tracking hospital-associated multidrug-resistant bacteria, Cell Mol. Life Sci., 2021, vol. 78, no. 6, pp. 2585‒2606.
Callaway, T.R., Carr, M.A., Edrington, T.S., Anderson, R.C., and Nisbet, D.J., Diet, Escherichia coli O157:H7, and cattle: a review after 10 years, Curr. Iss. Mol. Biol., 2009, vol. 11, no. 2, pp. 67‒79.
de Oliveira, G.A. and Silva, J.L., Cryo-EM to visualize the structural organization of viruses, Curr. Opin. Virol., 2021, vol. 49, pp. 86‒91.
Freedman, S.B., Xie, J., Neufeld, M.S., Hamilton, W.L., Hartling, L., Tarr, P.I., Alberta Provincial Pediatric Enteric Infection Team, Nettel-Aguirre, A., Chuck, A., Lee, B., Johnson, D., Currie, G., Talbot, J., Jiang, J., Dickinson, J., et al., Shiga toxin-producing Escherichia coli infection, antibiotics, and risk of developing hemolytic uremic syndrome: a meta-analysis, Clin. Infect. Dis., 2016, vol. 62, no. 10, pp. 1251‒1258.
Golomidova, A.K., Efimov, A.D., Kulikov, E.E., Kuznetsov, A.S., Belalov, I.S., and Letarov, A.V., O antigen restricts lysogenization of non-O157 Escherichia coli strains by Stx-converting bacteriophage phi24B, Sci. Rep., 2021, vol. 11, no. 1, p. 3035.
Kulikov, E.E., Golomidova, A.K., Prokhorov, N.S., Ivanov, P.A., and Letarov, A.V., High-throughput LPS profiling as a tool for revealing of bacteriophage infection strategies, Sci. Rep., 2019, vol. 9, no. 1, p. 2958.
Llarena, A.K., Aspholm, M., O’Sullivan, K., Wegrzyn, G., and Lindback, T., Replication region analysis reveals non-lambdoid Shiga toxin converting bacteriophages, Front. Microbiol., 2021, vol. 12, p. 640945.
Mathieu, A., Dion, M., Deng, L., Tremblay, D., Moncaut, E., Shah, S.A., Stokholm, J., Krogfelt, K.A., Schjorring, S., Bisgaard, H., Nielsen, D.S., Moineau, S., and Petit, M.A., Virulent coliphages in 1-year-old children fecal samples are fewer, but more infectious than temperate coliphages, Nat. Commun., 2020, vol. 11, no. 1, p. 378.
Muniesa, M., Serra-Moreno, R., and Jofre, J., Free Shiga toxin bacteriophages isolated from sewage showed diversity although the stx genes appeared conserved, Environ. Microbiol., 2004, vol. 6, no. 7, pp. 716‒725.
Riley, L.W. Pandemic lineages of extraintestinal pathogenic Escherichia coli, Clin. Microbiol. Infect., 2014, vol. 20, no. 5, pp. 380‒390.
Rollauer, S.E., Sooreshjani, M.A., Noinaj, N., and Buchanan, S.K., Outer membrane protein biogenesis in gram-negative bacteria, Philos. Trans. R. Soc. Lond. B Biol. Sci., 2015, vol. 370, no. 1679.
Sausset, R., Petit, M.A., Gaboriau-Routhiau, V., and De Paepe, M., New insights into intestinal phages, Mucosal Immunol., 2020, vol. 13, no. 2, pp. 205‒215.
Smith, D.L., James, C.E., Sergeant, M.J., Yaxian, Y., Saunders, J.R., McCarthy, A.J., and Allison, H.E., Short-tailed stx phages exploit the conserved YaeT protein to disseminate Shiga toxin genes among enterobacteria, J. Bacteriol., 2007, vol. 189, no. 20, pp. 7223‒7233.
Smith, D.L., Rooks, D.J., Fogg, P.C., Darby, A.C., Thomson, N.R., McCarthy, A.J., and Allison, H.E., Comparative genomics of Shiga toxin encoding bacteriophages, BMC Genomics, 2012, vol. 13, p. 311.
Todd, E.P.D., Preliminary estimates of costs of foodborne disease in Canada and costs to reduce salmonellosis, J. Food Prot., 1989, vol. 52, no. 8, pp. 586‒594.
Trachtman, H., Austin, C., Lewinski, M., and Stahl, R.A., Renal and neurological involvement in typical Shiga toxin-associated HUS, Nat. Rev. Nephrol., 2012, vol. 8, no. 11, pp. 658‒669.
Zhang, J.T., Yang, F., Du, K., Li, W.F., Chen, Y., Jiang, Y.L., Li, Q., and Zhou, C.Z., Structure and assembly pattern of a freshwater short-tailed cyanophage Pam1, Structure, 2022, vol. 30, no. 2, pp. 240‒251, e244.
Funding
This work was supported by ongoing institutional funding. No additional grants to carry out or direct this particular research were obtained.
Author information
Authors and Affiliations
Corresponding author
Ethics declarations
ETHICS APPROVAL AND CONSENT TO PARTICIPATE
This work does not contain any studies involving human and animal subjects.
CONFLICT OF INTEREST
The authors of this work declare that they have no conflicts of interest.
Additional information
Translated by E. Makeeva
Publisher’s Note.
Pleiades Publishing remains neutral with regard to jurisdictional claims in published maps and institutional affiliations.
Rights and permissions
About this article
Cite this article
Kuznetsov, A.S., Moiseenko, A.V., Kulikov, E.E. et al. CryoEM Investigation of Three-Dimentional Structure of the Stx-Converting Bacteriophage ϕ24B. Microbiology 93, 349–352 (2024). https://doi.org/10.1134/S0026261723604591
Received:
Revised:
Accepted:
Published:
Issue Date:
DOI: https://doi.org/10.1134/S0026261723604591