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Structure of phage P22 cell envelope–penetrating needle

Abstract

Bacteriophage P22 infects Salmonella enterica by injecting its genetic material through the cell envelope. During infection, a specialized tail needle, gp26, is injected into the host, likely piercing a hole in the host cell envelope. The 2.1-Å crystal structure of gp26 reveals a 240-Å elongated protein fiber formed by two trimeric coiled-coil domains interrupted by a triple β-helix. The N terminus of gp26 plugs the portal protein channel, retaining the genetic material inside the virion. The C-terminal tip of the fiber exposes β-hairpins with hydrophobic tips similar to those seen in class II fusion peptides. The α-helical core connecting these two functionally polarized tips presents four trimerization octads with consensus sequence IXXLXXXV. The slender conformation of the gp26 fiber minimizes the surface exposed to solvent, which is consistent with the idea that gp26 traverses the cell envelope lipid bilayers.

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Figure 1: Three-dimensional structure of phage P22 tail needle gp26.
Figure 2: Helical core of gp26.
Figure 3: Comparative analysis of gp26 helical core (domain II) and the ectodomain of influenza hemagglutinin.
Figure 4: The basic C-terminal tip of gp26 may be involved in membrane penetration.
Figure 5: Fitting of tail needle gp26 into the asymmetric cryo-EM reconstruction of mature P22 phage.

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References

  1. Lander, G.C. et al. The structure of an infectious p22 virion shows the signal for headful DNA packaging. Science 312, 1791–1795 (2006).

    Article  CAS  Google Scholar 

  2. Chang, J., Weigele, P., King, J., Chiu, W. & Jiang, W. Cryo-EM asymmetric reconstruction of bacteriophage P22 reveals organization of its DNA packaging and infecting machinery. Structure 14, 1073–1082 (2006).

    Article  CAS  Google Scholar 

  3. Strauss, H. & King, J. Steps in the stabilization of newly packaged DNA during phage P22 morphogenesis. J. Mol. Biol. 172, 523–543 (1984).

    Article  CAS  Google Scholar 

  4. Lenk, E., Casjens, S., Weeks, J. & King, J. Intracellular visualization of precursor capsids in phage P22 mutant infected cells. Virology 68, 182–199 (1975).

    Article  CAS  Google Scholar 

  5. Berget, P.B. & Poteete, A.R. Structure and functions of the bacteriophage P22 tail protein. J. Virol. 34, 234–243 (1980).

    CAS  PubMed  PubMed Central  Google Scholar 

  6. Molineux, I.J. No syringes please, ejection of phage T7 DNA from the virion is enzyme driven. Mol. Microbiol. 40, 1–8 (2001).

    Article  CAS  Google Scholar 

  7. Israel, V. A model for the adsorption of phage P22 to Salmonella typhimurium. J. Gen. Virol. 40, 669–673 (1978).

    Article  CAS  Google Scholar 

  8. Bhardwaj, A., Olia, A.S., Walker-Kopp, N. & Cingolani, G. Domain organization and polarity of tail needle GP26 in the portal vertex structure of bacteriophage P22. J. Mol. Biol. 371, 374–387 (2007).

    Article  CAS  Google Scholar 

  9. Andrews, D. et al. Bacteriophage P22 tail accessory factor GP26 is a long triple-stranded coiled-coil. J. Biol. Chem. 280, 5929–5933 (2005).

    Article  CAS  Google Scholar 

  10. Israel, V. E proteins of bacteriophage P22. I. Identification and ejection from wild-type and defective particles. J. Virol. 23, 91–97 (1977).

    CAS  PubMed  PubMed Central  Google Scholar 

  11. Moak, M. & Molineux, I.J. Peptidoglycan hydrolytic activities associated with bacteriophage virions. Mol. Microbiol. 51, 1169–1183 (2004).

    Article  CAS  Google Scholar 

  12. Cingolani, G., Andrews, D. & Casjens, S. Crystallogenesis of bacteriophage P22 tail accessory factor gp26 at acidic and neutral pH. Acta Crystallogr. Sect. F Struct. Biol. Cryst. Commun. 62, 477–482 (2006).

    Article  CAS  Google Scholar 

  13. Bullough, P.A., Hughson, F.M., Skehel, J.J. & Wiley, D.C. Structure of influenza haemagglutinin at the pH of membrane fusion. Nature 371, 37–43 (1994).

    Article  CAS  Google Scholar 

  14. Eckert, D.M. & Kim, P.S. Mechanisms of viral membrane fusion and its inhibition. Annu. Rev. Biochem. 70, 777–810 (2001).

    Article  CAS  Google Scholar 

  15. Crick, F.H.C. The packing of α-helices: simple coiled-coils. Acta Crystallogr. 6, 689–697 (1953).

    Article  CAS  Google Scholar 

  16. Burkhard, P., Kammerer, R.A., Steinmetz, M.O., Bourenkov, G.P. & Aebi, U. The coiled-coil trigger site of the rod domain of cortexillin I unveils a distinct network of interhelical and intrahelical salt bridges. Structure 8, 223–230 (2000).

    Article  CAS  Google Scholar 

  17. Stummeyer, K., Dickmanns, A., Muhlenhoff, M., Gerardy-Schahn, R. & Ficner, R. Crystal structure of the polysialic acid-degrading endosialidase of bacteriophage K1F. Nat. Struct. Mol. Biol. 12, 90–96 (2005).

    Article  CAS  Google Scholar 

  18. Tang, L., Marion, W.R., Cingolani, G., Prevelige, P.E. & Johnson, J.E. Three-dimensional structure of the bacteriophage P22 tail machine. EMBO J. 24, 2087–2095 (2005).

    Article  CAS  Google Scholar 

  19. Modis, Y., Ogata, S., Clements, D. & Harrison, S.C. Structure of the dengue virus envelope protein after membrane fusion. Nature 427, 313–319 (2004).

    Article  CAS  Google Scholar 

  20. Evilevitch, A., Lavelle, L., Knobler, C.M., Raspaud, E. & Gelbart, W.M. Osmotic pressure inhibition of DNA ejection from phage. Proc. Natl. Acad. Sci. USA 100, 9292–9295 (2003).

    Article  CAS  Google Scholar 

  21. Kindt, J., Tzlil, S., Ben-Shaul, A. & Gelbart, W.M. DNA packaging and ejection forces in bacteriophage. Proc. Natl. Acad. Sci. USA 98, 13671–13674 (2001).

    Article  CAS  Google Scholar 

  22. Coombs, D.H. & Ariska, F. in Molecular Biology of Bacteriophage T4 (ed. Karam, J.D.) 259–281 (American Society of Microbiology, Washington DC, 1994).

    Google Scholar 

  23. Goldberg, E., Grinius, L. & Letellier, L. in Molecular Biology of Bacteriophage T4 (ed. Karam, J.D.) 347–356 (American Society of Microbiology, Washington DC, 1994).

    Google Scholar 

  24. Kanamaru, S. et al. Structure of the cell-puncturing device of bacteriophage T4. Nature 415, 553–557 (2002).

    Article  CAS  Google Scholar 

  25. Nakagawa, H., Arisaka, F. & Ishii, S. Isolation and characterization of the bacteriophage T4 tail-associated lysozyme. J. Virol. 54, 460–466 (1985).

    CAS  PubMed  PubMed Central  Google Scholar 

  26. Otwinowski, Z. & Minor, W. Processing of X-ray diffraction data collected in oscillation mode. Methods Enzymol. 276, 307–326 (1997).

    Article  CAS  Google Scholar 

  27. Collaborative Computational Project, Number 4. The CCP4 suite: programs for protein crystallography. Acta Crystallogr. D Biol. Crystallogr. 50, 760–763 (1994).

  28. Terwilliger, T.C. SOLVE and RESOLVE: automated structure solution and density modification. Methods Enzymol. 374, 22–37 (2003).

    Article  CAS  Google Scholar 

  29. Emsley, P. & Cowtan, K. Coot: model-building tools for molecular graphics. Acta Crystallogr. D Biol. Crystallogr. 60, 2126–2132 (2004).

    Article  Google Scholar 

  30. Brunger, A.T. et al. Crystallography & NMR system: a new software suite for macromolecular structure determination. Acta Crystallogr. D Biol. Crystallogr. 54, 905–921 (1998).

    Article  CAS  Google Scholar 

  31. Sheldrick, G.M. & Schneider, T.R. SHELXL: High-resolution refinement. Methods Enzymol. 277, 319–343 (1997).

    Article  CAS  Google Scholar 

  32. Larsen, N.A. et al. Structure determination of a cocaine hydrolytic antibody from a pseudomerohedrally twinned crystal. Acta Crystallogr. D Biol. Crystallogr. 58, 2055–2059 (2002).

    Article  Google Scholar 

  33. Yeates, T.O. & Fam, B.C. Protein crystals and their evil twins. Structure 7, R25–R29 (1999).

    Article  CAS  Google Scholar 

  34. Pettersen, E.F. et al. UCSF Chimera–a visualization system for exploratory research and analysis. J. Comput. Chem. 25, 1605–1612 (2004).

    Article  CAS  Google Scholar 

  35. Merritt, E.A. & Murphy, M.E. Raster3D Version 2.0. A program for photorealistic molecular graphics. Acta Crystallogr. D Biol. Crystallogr. 50, 869–873 (1994).

    Article  CAS  Google Scholar 

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Acknowledgements

We thank N. Walker for technical help. We are grateful to V. Stojanoff at the NSLS and to the macCHESS staff for beam time and assistance in data collection. This work was supported in part by US National Science Foundation grant MCB-990526 to S.C.

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Authors

Contributions

A.S.O. and G.C. contributed to the structural and biochemical studies. S.C. participated in the design and coordination of the study. A.S.O., S.C. and G.C. wrote the manuscript.

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Correspondence to Gino Cingolani.

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Supplementary Figures 1–3 and Supplementary Table 1 (PDF 4534 kb)

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Olia, A., Casjens, S. & Cingolani, G. Structure of phage P22 cell envelope–penetrating needle. Nat Struct Mol Biol 14, 1221–1226 (2007). https://doi.org/10.1038/nsmb1317

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