Tailed bacteriophages: Structure, Function and Evolution A tale of two tails Michael Rossmann Purdue

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Tailed bacteriophages Structure, Function and
Evolution(A tale of two tails) Michael
RossmannPurdue University
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f29
T4
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T4 168 kbp genome 300 ORFs 40 structural
proteins Host E. Coli Gram negative Cell wall
inside periplasmic space Packages T4 concatomer
Phi29 20 kbp genome 20 ORFs 8 structural
proteins 1 structural RNA Host B.
Subtilis Gram positive Cell wall external to
single cell membrane Packeges DNAgp3 genome
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Bacteriophage phi29
Rossmann Lab Valorie Bowman Paul Chipman Kay
Choi Marc Morais Alan Simpson Yizhi Tao Ye Xiang
Anderson Lab Mohammed Badasso Shelley Grimes Paul
Jardine Jaya Kotl
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Tao et al Cell, 1998, 95431-437
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Three layers of DNA
DNA ring Connector appendages
gp3? head fiber bulge lower collar knob
?
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Phi29 head-tail head-tail Connector
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f29 dodecameric connector
Simpson et al, Nature, 2000, 408745-750
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pRNA difference map
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Partially packaged prohead with pRNA and ATPase
(gp16)
Empty Prohead with pRNA
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Connector
pRNA pentamer Difference density
Gp16 ATPase
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Phage F29 DNA packaging motor
Simpson et al, Nature, 2000, 408745-750
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The T4 structure
Purdue University Anthoni Batisti Paul
Chipman Andrei Fokine Victor Kostyuchenko Pertr
Leiman
Moscow Vadim Mesyanzhinov Mikhail Schneider
Catholic University of America Venigella Rao
Tokyo Institute of Technology Fumio Arisaka Shuji
Kanamaru
NIH Bethesda Bijan Ahvazi Karen Boeshans Alasdair
Steven
University of Maryland Lindsay Black
Santiago Mark van Raaij
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Too complex to crystallize Bacteriophage T4 an
efficient infection machine
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Phage particle assembly
Head
Long tail fibers
Fibritin
gp13, gp14, gp2, gp4
gp63
Tail
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T4 Tail assembly
Wedge
25, 6
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53, 7, 8
10, 11
gp9
15S
gp12
gp48 gp54
120S
Hub
70S
5, 27
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22S
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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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Common topologies
gp11
gp10
gp12
Interaction in 3D
X
big insertion
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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
Kanamaru et al, Nature 2002, 415553-557
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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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Baseplate movie
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48 54
6 25 53
9
8
27
7
5
10
11
12
26?
Kostyuchenko et al, Nat. Struct. Biol. 2003,
10688-693
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Hexagonal Baseplate
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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

Leiman et al, Cell 2004, 118419-429
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Contracted tail movie
Contracted tail Structure
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13 14
15
3
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9
8
7
11
6 25 53
10
12
19
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Extended sheath 1 turn
Contracted sheath 2 turns
Kostyuchenko et al, Nat.Struct, Mol. Biol. 2005
in press
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November 18th, 2004
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Movie