Tailed bacteriophages: Structure, Function and Evolution
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Tailed bacteriophages: Structure, Function and Evolution (A tale of two tails) Michael Rossmann Purdue University. φ 29. T4. 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

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Tailed bacteriophages: Structure, Function and Evolution(A tale of two tails)Michael RossmannPurdue University


φ29

T4


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


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



Three layers of DNA

DNA ring

Connector

appendages

gp3?

head fiber

bulge

lower collar

knob

?



φ29 dodecameric connector

Simpson et al,Nature, 2000,408:745-750



Partially packaged prohead

with pRNA and ATPase (gp16)

Empty Prohead

with pRNA


Connector

pRNA pentamer

Difference density

Gp16 ATPase


Phage Φ29:

DNA packaging motor

Simpson et al,Nature, 2000,408:745-750


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


Too complex to crystallize bacteriophage t4 an efficient infection machine
Too complex to crystallize: Bacteriophage T4an efficient infection machine


Phage particle assembly
Phage particle assembly

Head

Long tail fibers

Fibritin

gp13, gp14, [gp2], [gp4]

[gp63]

Tail


T4 tail assembly

Tail

terminator

gp15

138 gp18

138 gp19

T4 Tail assembly

Wedge

6

25, 6

53, 7, 8

10, 11

gp9

15S

gp12

gp48

gp54

120S

Hub

70S

5, 27

29

22S


Crystal structures of the baseplate proteins
Crystal structures of the baseplate proteins

gp9,

LTF attachment

gp11,

STF attachment

gp8

gp12

C-term

gp12, STF

gp5-gp27

30 nm


Common topologies
Common topologies

gp11

gp10

gp12

Interaction in 3D

X

big insertion


Gp5 gp27 complex structure overview

80 Å

Gp5-gp27 complex structure overview

gp27 trimer

gp5 N-terminal domain

190 Å

gp5 lysozymedomain

gp5 C-terminal

domain

Kanamaru et al, Nature 2002,415:553-557


Hexagonal conformation tube baseplates
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


Baseplate movie
Baseplate movie

19

48 54

6 25 53

9

27

8

7

5

10

11

12

26?

Kostyuchenko et al, Nat. Struct. Biol. 2003,10:688-693



Star shaped baseplates contracted tails
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, 118:419-429


Contracted tail movie

Contracted tail

Structure

Contracted tail movie

23

20

13 14

15

3

19

18

34

9

7

8

6 25 53

11

10

12

19


Extended sheath

1 turn

Contracted sheath

2 turns

Kostyuchenko et al, Nat.Struct, Mol. Biol. 2005 in press


November 18th, 2004



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