φ 29
description
Transcript of φ 29
Tailed bacteriophages: Structure, Function and Evolution
(A tale of two tails)
Michael RossmannPurdue University
T4
φ29
T4168 kbp genome
~300 ORFs
40 structural proteins
Host: E. Coli
Gram negativeCell wall inside periplasmic
space
Packages T4 concatomer
Phi2920 kbp genome
~20 ORFs
8 structural proteins1 structural RNA
Host: B. Subtilis
Gram positiveCell wall external to single
cell membrane
Packeges DNAgp3 genome
Bacteriophage phi29
Rossmann LabValorie Bowman
Paul ChipmanKay Choi
Marc MoraisAlan Simpson
Yizhi TaoYe Xiang
Anderson LabMohammed Badasso
Shelley GrimesPaul Jardine
Jaya Kotl
Tao et al Cell, 1998, 95:431-
437
gp3?
head fiber
bulge
lower collar
knob
DNA ring
Connector
appendages
?
Three layers of DNA
Phi29 head-tail head-tail Connector
Simpson et al, Nature, 2000, 408:745-750
φ29 dodecameric connector
pRNA difference map
Partially packaged proheadwith pRNA and ATPase
(gp16)
Empty Proheadwith pRNA
Connector
pRNA pentamerDifference density
Gp16 ATPase
Phage Φ29:DNA packaging motor
Simpson et al, Nature, 2000, 408:745-750
The T4 structurePurdue University
Anthoni BatistiPaul ChipmanAndrei Fokine
Victor KostyuchenkoPertr Leiman
MoscowVadim Mesyanzhinov
Mikhail Schneider
Catholic University of America
Venigella RaoTokyo Institute of
TechnologyFumio Arisaka
Shuji Kanamaru
University of MarylandLindsay Black
NIH BethesdaBijan Ahvazi
Karen BoeshansAlasdair Steven
Santiago Mark van Raaij
Too complex to crystallize: Bacteriophage T4 an efficient infection machine
Phage particle assemblyHead
Tail
Fibritin
gp13, gp14, [gp2], [gp4]
[gp63]
Long tail
fibers
138 gp19 138 gp18
5, 27
T4 Tail assembly
53, 7, 810, 11
25, 6
15S
29
22S
Wedge
Hub
70S
6
120S
Tailterminator
gp15
gp9
gp12 gp48gp54
Crystal structures of the baseplate proteins
gp8gp11,STF attachment
gp9,LTF
attachment
gp5-gp27
gp12C-term
30 nm
gp12, STF
Common topologiesgp11 gp10 gp12
Interaction in 3DX big insertion
Gp5-gp27 complex structure overviewgp27 trimer
gp5 N-terminal domain
gp5 lysozyme domain
gp5 C-terminaldomain
80 Å
190 Å
Kanamaru et al, Nature 2002, 415:553-557
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 m
• 12 Å resolution
Baseplate movie
19
48 54
6 25 53
98
7
10
11 12
5
26?
27
Kostyuchenko et al, Nat. Struct. Biol. 2003, 10:688-693
Hexagonal Baseplate
Star-shaped baseplates (contracted tails)
• Sixfold symmetry
• 1580 particles used in the reconstruction
• Defoci 0.7 – 3.3 m
• 17 Å resolution
Leiman et al, Cell 2004,
118:419-429
Contracted tail movie
19
18
987
10
11
12
23
34
15
2013 14
319
6 25 53
Contracted tail
Structure
Extended sheath1 turn
Contracted sheath2 turns
Kostyuchenko et al, Nat.Struct, Mol. Biol. 2005
in press
November 18th, 2004
Movie