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Structure of the cell-puncturing device of bacteriophage T4

Nature volume 415, pages 553–557 (2002)Cite this article

Abstract

Bacteriophage T4 has a very efficient mechanism for infecting cells1. The key component of this process is the baseplate, located at the end of the phage tail, which regulates the interaction of the tail fibres and the DNA ejection machine2. A complex of gene product (gp) 5 (63K) and gp27 (44K), the central part of the baseplate, is required to penetrate the outer cell membrane of Escherichia coli and to disrupt the intermembrane peptidoglycan layer, promoting subsequent entry of phage DNA into the host. We present here a crystal structure of the (gp5–gp27)3 321K complex, determined to 2.9 Å resolution and fitted into a cryo-electron microscopy map at 17 Å resolution of the baseplate-tail tube assembly. The carboxy-terminal domain of gp5 is a triple-stranded β-helix that forms an equilateral triangular prism, which acts as a membrane-puncturing needle. The middle lysozyme domain of gp5, situated on the periphery of the prism, serves to digest the peptidoglycan layer. The amino-terminal, antiparallel β-barrel domain of gp5 is inserted into a cylinder formed by three gp27 monomers, which may serve as a channel for DNA ejection.

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Figure 1: Assembly of (gp27–gp5*–gp5C)3.
Figure 2: Structure of the (gp27–gp5*–gp5C)3 complex.
Figure 3: CryoEM reconstruction of the T4 baseplate-tail tube assembly.
Figure 4: Structural comparison with T4L13 shows that residues Pro 363-Ala 364-Asp 365 of linker 2 of the three-fold-related neighbouring subunit bind, correctly oriented, into three of the four peptide-binding sites used by the peptidoglycan substrate.

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Acknowledgements

We thank T. S. Baker for establishing the electron microscopy facilities at Purdue, where we collected the cryoEM data, A. A. Simpson for advice and help in data collection, and B. W. Matthews for discussion of the results. We thank the staff of BioCARS for their help and advice in the data collection at the Advanced Photon Source beam lines 14-BM-C and 14-BM-D. We thank S. Wilder for help in preparation of the manuscript. The work was supported by a National Science Foundation grant to M.G.R.; Grants-in-Aid for Scientific Research from the Ministry of Education, Science, Sports, and Culture of Japan to F.A.; a Howard Hughes Medical Institute grant to V.V.M.; a reinvestment grant from Purdue University; and a Keck Foundation award for the purchase of a Philips CM300 electron microscope.

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  1. Shuji Kanamaru and Petr G. Leiman: These authors contributed equally to this work

Authors and Affiliations

  1. Department of Biological Sciences, Purdue University, West Lafayette, 47907-1392, Indiana, USA

    Shuji Kanamaru, Petr G. Leiman, Victor A. Kostyuchenko, Paul R. Chipman & Michael G. Rossmann

  2. Laboratory of Molecular Bioengineering, Shemyakin–Ovchinnikov Institute of Bioorganic Chemistry, 16/10 Miklukho-Maklaya Street, Moscow, 117997, Russia

    Victor A. Kostyuchenko & Vadim V. Mesyanzhinov

  3. Department of Life Science, Faculty of Bioscience and Biotechnology, Tokyo Institute of Technology, 4259 Nagatsuta, Midori-ku, 226-8501, Yokohama, Japan

    Fumio Arisaka

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  1. Shuji Kanamaru
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Correspondence to Michael G. Rossmann.

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Kanamaru, S., Leiman, P., Kostyuchenko, V. et al. Structure of the cell-puncturing device of bacteriophage T4. Nature 415, 553–557 (2002). https://doi.org/10.1038/415553a

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