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