Structure of the Bacteriophage T4 Contracted Tail

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Guy and Eleanors unknown past A story with a
happy ending
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1962 Guy marries a read head from a former
English penal colony
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• November 5th 1962
• Guy thinks he is Guy Fawkes. Teaches our 3
  children pyrotechnics on a foggy night in
  Cambridge
• Is suspect for stealing City of Cambridge lights
  to decorate his identical twin brothers car. His
  brother is jailed, Guy goes free

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1966 Guy arrested by Purdue police for trying to
smuggle a bomb onto an airplane. I go to the
police station and am able to procure his release
on guarantee of his good behavior. The bomb
turned out to be a boxed insulin model that
Guy refused to check with his other luggage.
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1968 Guy is not a real New Zealander after
all! On a visit to Oxford I have the good
fortune of meeting Guys brother-in-law. Guys
brother-in-law tells me, with a good deal of
indignation. that only he and not Guy, is a true
New Zealander.
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Dodson, E., M. M. Harding, D. C. Hodgkin, M. G.
Rossmann. 1966. The crystal structure of
insulin. III. Evidence for a 2-fold axis in
rhombohedral zinc insulin. J. Mol. Biol.
16227-241. Rossmann, M. G., D. M. Blow, M. M.
Harding, E. Coller. 1964. The relative
positions of independent molecules within the
same asymmetric unit. Acta Crystallogr.
17338-342.
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The efficient infection machine of bacteriophage
T4 Where crystallography meets electron
microscopyMichael RossmannPurdue University
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Cryo-EM Micrographs
Mature Dengue Virus
Immature Dengue Virus
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Dengue Virus Membrane proteins in situ
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E glycoprotein dimer neutral pH
TBEV F. Rey et al Nature, 1995, 375, 291-298
Dengue Y. Modis et al PNAS, 2003, 100,
6986-6991 Y. Zhang et al, Structure 2004,
in press
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An efficient T4 bacteriophage infection
machine dsDNA genome, 168kbp, 300 ORFs, 40
structural proteinsHost E. Coli
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T4 head movie
T4 phage head-tail symmetry mismatch
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Tail assembly
Wedge
25, 6
6
53, 7, 8
10, 11
gp9
15S
gp12
gp48 gp54
120S
Hub
70S
5, 27
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22S
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Phage particle assembly
Head
Long tail fibers
Fibritin
gp13, gp14, gp2, gp4
gp63
Tail
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Crystal structures of the baseplate proteins
gp9, LTF attachment
gp11, STF attachment
gp8
gp12 C-term
gp12, STF
gp5-gp27
30 nm
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Gp5-gp27 complex structure overview
80 Å
gp27 trimer
gp5 N-terminal domain
190 Å
gp5 lysozyme domain
gp5 C-terminal domain
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Hexagonal conformation (tube-baseplates)

• Initial model hexagonal prism connected to a
  tube
• Sixfold symmetry
• 945 particles used in the reconstruction
• Defoci 1.5 3.5 mm
• 12 Å resolution

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Uniqueness of location of proteins with known
crystal structures

• Exhaustive and local 6D searches for the best
  match between a crystal structure and the cryoEM
  map (COLORES and EMfit)
• COLORES score map Each grid point corresponds to
  the best correlation coefficient obtained at this
  position for all orientations of the fitted gp8
  dimer

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Baseplate movie
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48 54
6 25 53
9
8
27
7
5
10
11
12
26?
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The baseplate regulates the infection process
Hexagon
Star
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Star-shaped baseplates (contracted tails)

• Sixfold symmetry
• 1580 particles used in the reconstruction
• Defoci 0.7 3.3 mm
• 17 Å resolution

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Contracted tail movie
Contracted tail Structure
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13 14
15
3
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34
9
8
7
11
6 25 53
10
12
19
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Extended sheath 1 turn
Contracted sheath 2 turns
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Infection process

• The conformational switch is triggered by
• Interaction of the STFs with LPSs
• Interaction of the LTFs with gp11s

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Movie
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Acknowledgements
T4 Petr Leiman, Victor Kostyuchenko, Paul
Chipman, Shuji Kanamaru, Mark van Raaij, Andrei
Fukin, Misha Schneider, Fumio Arisaka, Venigalla.
Rao, Vadim Mesynanzhinov Dengue Virus Wei Zhang,
Ying Zhang, Suchetana Mukhopadhyay, Paul Chipman,
Tim Baker, Jim Strauss, Richard Kuhn