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Title: Structural basis of superinfection exclusion by bacteriophage T4 Spackle

Abstract

A bacterial cell infected with T4 phage rapidly establishes resistance against further infections by the same or closely related T-even-type bacteriophages – a phenomenon called superinfection exclusion. Here we show that one of the T4 early gene products and a periplasmic protein, Spackle, forms a stoichiometric complex with the lysozyme domain of T4 tail spike protein gp5 and potently inhibits its activity. Crystal structure of the Spackle-gp5 lysozyme complex shows that Spackle binds to a horseshoe-shaped basic patch surrounding the oligosaccharide-binding cleft and induces an allosteric conformational change of the active site. In contrast, Spackle does not appreciably inhibit the lysozyme activity of cytoplasmic T4 endolysin responsible for cell lysis to release progeny phage particles at the final step of the lytic cycle. Our work reveals a unique mode of inhibition for lysozymes, a widespread class of enzymes in biology, and provides a mechanistic understanding of the T4 bacteriophage superinfection exclusion.

Authors:
 [1];  [1];  [1];  [1];  [2]; ORCiD logo [1]
  1. Univ. of Minnesota, Minneapolis, MN (United States)
  2. Cornell Univ., Ithaca, NY (United States)
Publication Date:
Research Org.:
Argonne National Lab. (ANL), Argonne, IL (United States)
Sponsoring Org.:
USDOE Office of Science (SC), Basic Energy Sciences (BES). Scientific User Facilities Division
Contributing Org.:
Advanced Photon Source (APS), Argonne National Laboratory (ANL), Argonne, IL (US)
OSTI Identifier:
1729706
Grant/Contract Number:  
AC02-06CH11357
Resource Type:
Accepted Manuscript
Journal Name:
Communications Biology
Additional Journal Information:
Journal Volume: 3; Journal Issue: 1; Journal ID: ISSN 2399-3642
Publisher:
Springer Nature
Country of Publication:
United States
Language:
ENGLISH
Subject:
59 BASIC BIOLOGICAL SCIENCES

Citation Formats

Shi, Ke, Oakland, Justin T., Kurniawan, Fredy, Moeller, Nicholas H., Banerjee, Surajit, and Aihara, Hideki. Structural basis of superinfection exclusion by bacteriophage T4 Spackle. United States: N. p., 2020. Web. https://doi.org/10.1038/s42003-020-01412-3.
Shi, Ke, Oakland, Justin T., Kurniawan, Fredy, Moeller, Nicholas H., Banerjee, Surajit, & Aihara, Hideki. Structural basis of superinfection exclusion by bacteriophage T4 Spackle. United States. https://doi.org/10.1038/s42003-020-01412-3
Shi, Ke, Oakland, Justin T., Kurniawan, Fredy, Moeller, Nicholas H., Banerjee, Surajit, and Aihara, Hideki. Thu . "Structural basis of superinfection exclusion by bacteriophage T4 Spackle". United States. https://doi.org/10.1038/s42003-020-01412-3. https://www.osti.gov/servlets/purl/1729706.
@article{osti_1729706,
title = {Structural basis of superinfection exclusion by bacteriophage T4 Spackle},
author = {Shi, Ke and Oakland, Justin T. and Kurniawan, Fredy and Moeller, Nicholas H. and Banerjee, Surajit and Aihara, Hideki},
abstractNote = {A bacterial cell infected with T4 phage rapidly establishes resistance against further infections by the same or closely related T-even-type bacteriophages – a phenomenon called superinfection exclusion. Here we show that one of the T4 early gene products and a periplasmic protein, Spackle, forms a stoichiometric complex with the lysozyme domain of T4 tail spike protein gp5 and potently inhibits its activity. Crystal structure of the Spackle-gp5 lysozyme complex shows that Spackle binds to a horseshoe-shaped basic patch surrounding the oligosaccharide-binding cleft and induces an allosteric conformational change of the active site. In contrast, Spackle does not appreciably inhibit the lysozyme activity of cytoplasmic T4 endolysin responsible for cell lysis to release progeny phage particles at the final step of the lytic cycle. Our work reveals a unique mode of inhibition for lysozymes, a widespread class of enzymes in biology, and provides a mechanistic understanding of the T4 bacteriophage superinfection exclusion.},
doi = {10.1038/s42003-020-01412-3},
journal = {Communications Biology},
number = 1,
volume = 3,
place = {United States},
year = {2020},
month = {11}
}

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Works referenced in this record:

Bacteriophage resistance mechanisms
journal, March 2010

  • Labrie, Simon J.; Samson, Julie E.; Moineau, Sylvain
  • Nature Reviews Microbiology, Vol. 8, Issue 5
  • DOI: 10.1038/nrmicro2315

Spackle and Immunity Functions of Bacteriophage T4
journal, January 1974


Structure of the cell-puncturing device of bacteriophage T4
journal, January 2002

  • Kanamaru, Shuji; Leiman, Petr G.; Kostyuchenko, Victor A.
  • Nature, Vol. 415, Issue 6871
  • DOI: 10.1038/415553a

A covalent enzyme-substrate intermediate with saccharide distortion in a mutant T4 lysozyme
journal, December 1993


Structure of the T4 baseplate and its function in triggering sheath contraction
journal, May 2016

  • Taylor, Nicholas M. I.; Prokhorov, Nikolai S.; Guerrero-Ferreira, Ricardo C.
  • Nature, Vol. 533, Issue 7603
  • DOI: 10.1038/nature17971

Isolation and characterization of the bacteriophage T4 tail-associated lysozyme.
journal, January 1985


Guards of the great wall: bacterial lysozyme inhibitors
journal, October 2012

  • Callewaert, Lien; Van Herreweghe, Joris M.; Vanderkelen, Lise
  • Trends in Microbiology, Vol. 20, Issue 10
  • DOI: 10.1016/j.tim.2012.06.005

Gene 61.3 of Bacteriophage T4 Is the spackle Gene
journal, August 1999


Translationsl regulation of expression of the bacteriophage T4 lysozyme gene
journal, January 1986

  • McPheeters, David S.; Christensen, Alan; Young, Elton T.
  • Nucleic Acids Research, Vol. 14, Issue 14
  • DOI: 10.1093/nar/14.14.5813

Deciphering key features in protein structures with the new ENDscript server
journal, April 2014

  • Robert, Xavier; Gouet, Patrice
  • Nucleic Acids Research, Vol. 42, Issue W1
  • DOI: 10.1093/nar/gku316

Structural remodeling of bacteriophage T4 and host membranes during infection initiation
journal, August 2015

  • Hu, Bo; Margolin, William; Molineux, Ian J.
  • Proceedings of the National Academy of Sciences, Vol. 112, Issue 35
  • DOI: 10.1073/pnas.1501064112

Lysis from without
journal, January 2011


Features and development of Coot
journal, March 2010

  • Emsley, P.; Lohkamp, B.; Scott, W. G.
  • Acta Crystallographica Section D Biological Crystallography, Vol. 66, Issue 4
  • DOI: 10.1107/S0907444910007493

The functions of the phage T4 immunity and spackle genes in genetic exclusion
journal, October 1988


Control of Bacteriophage T4 Tail Lysozyme Activity During the Infection Process
journal, March 2005

  • Kanamaru, Shuji; Ishiwata, Yasutaka; Suzuki, Toshiharu
  • Journal of Molecular Biology, Vol. 346, Issue 4
  • DOI: 10.1016/j.jmb.2004.12.042

Structure and evolution of the Ivy protein family, unexpected lysozyme inhibitors in Gram-negative bacteria
journal, April 2007

  • Abergel, C.; Monchois, V.; Byrne, D.
  • Proceedings of the National Academy of Sciences, Vol. 104, Issue 15
  • DOI: 10.1073/pnas.0611019104

Structural Basis for the Inhibition of Human Lysozyme by PliC from Brucella abortus
journal, November 2013

  • Um, Si-Hyeon; Kim, Jin-Sik; Kim, Kuglae
  • Biochemistry, Vol. 52, Issue 51
  • DOI: 10.1021/bi401241c

Superinfection exclusion by T-even-type coliphages
journal, April 1994


The role of phage lysozyme in the life cycle of phage T4
journal, November 1968


Baseplate protein of bacteriophage T4 with both structural and lytic functions.
journal, January 1980


Macromolecular structure determination using X-rays, neutrons and electrons: recent developments in Phenix
journal, October 2019

  • Liebschner, Dorothee; Afonine, Pavel V.; Baker, Matthew L.
  • Acta Crystallographica Section D Structural Biology, Vol. 75, Issue 10
  • DOI: 10.1107/S2059798319011471

Lysis of T4-infected bacteria in the absence of lysozyme
journal, May 1968


Structural basis for the recognition of lysozyme by MliC, a periplasmic lysozyme inhibitor in Gram-negative bacteria
journal, January 2009

  • Yum, Soohwan; Kim, Moon Jong; Xu, Yongbin
  • Biochemical and Biophysical Research Communications, Vol. 378, Issue 2
  • DOI: 10.1016/j.bbrc.2008.11.039

Mapping of functional sites on the primary structure of the tail lysozyme of bacteriophage T4 by mutational analysis
journal, May 1998

  • Takeda, Shigeki; Hoshida, Kotaro; Arisaka, Fumio
  • Biochimica et Biophysica Acta (BBA) - Protein Structure and Molecular Enzymology, Vol. 1384, Issue 2
  • DOI: 10.1016/S0167-4838(98)00016-8

Characterization of the interactions between Escherichia coli receptors, LPS and OmpC, and bacteriophage T4 long tail fibers
journal, June 2016

  • Washizaki, Ayaka; Yonesaki, Tetsuro; Otsuka, Yuichi
  • MicrobiologyOpen, Vol. 5, Issue 6
  • DOI: 10.1002/mbo3.384

The structure of the proteinaceous inhibitor PliI from Aeromonas hydrophila in complex with its target lysozyme
journal, January 2015

  • Leysen, Seppe; Van Herreweghe, Joris M.; Yoneda, Kazunari
  • Acta Crystallographica Section D Biological Crystallography, Vol. 71, Issue 2
  • DOI: 10.1107/S1399004714025863

Bacteriophage T4 resistance to lysis-inhibition collapse
journal, August 1999


Structural basis of bacterial defense against g-type lysozyme-based innate immunity
journal, October 2012

  • Leysen, S.; Vanderkelen, L.; Weeks, S. D.
  • Cellular and Molecular Life Sciences, Vol. 70, Issue 6
  • DOI: 10.1007/s00018-012-1184-1

Roles of bacteriophage T4 gene 5 and gene s products in cell lysis.
journal, January 1980


Targeting mechanisms of tailed bacteriophages
journal, August 2018

  • Nobrega, Franklin L.; Vlot, Marnix; de Jonge, Patrick A.
  • Nature Reviews Microbiology, Vol. 16, Issue 12
  • DOI: 10.1038/s41579-018-0070-8

XDS
journal, January 2010

  • Kabsch, Wolfgang
  • Acta Crystallographica Section D Biological Crystallography, Vol. 66, Issue 2
  • DOI: 10.1107/S0907444909047337

The C-Terminal Fragment of the Precursor Tail Lysozyme of Bacteriophage T4 Stays as a Structural Component of the Baseplate after Cleavage
journal, May 1999


Crystallographic determination of the mode of binding of oligosaccharides to T4 bacteriophage lysozyme: Implications for the mechanism of catalysis
journal, April 1981


Phaser crystallographic software
journal, July 2007

  • McCoy, Airlie J.; Grosse-Kunstleve, Ralf W.; Adams, Paul D.
  • Journal of Applied Crystallography, Vol. 40, Issue 4
  • DOI: 10.1107/S0021889807021206

Morphogenesis of the T4 tail and tail fibers
journal, December 2010

  • Leiman, Petr G.; Arisaka, Fumio; van Raaij, Mark J.
  • Virology Journal, Vol. 7, Issue 1
  • DOI: 10.1186/1743-422X-7-355