Structural analysis of the bacteriophage T3 head-to-tail connector.
The connector protein of bacteriophage T3, p8, has been overexpressed in Escherichia coli. Purification of the oligomers built by several copies of p8 reveals a mixed population of dodecamers and tridecamers. The percentages of these two types of oligomers differ in every culture growth, indicating that assembly of this protein depends upon the conditions of the expression system. Those cultures that generated a majority of dodecamers allowed, after purification of the connectors, the two-dimensional crystallization of the dodecamers in a tetragonal arrangement, while the tridecamers did not form crystals. The processing and averaging of several images of frozen-hydrated crystals and their internal phase comparison shows that the crystals are arranged in a P42(1)2 space group, with cell unit dimensions of 165 x 165 A. The three-dimensional reconstruction generated with images of crystals ranging from 0 degrees to 60 degrees tilt reveals a wide domain surrounded by 12 protrusions and a narrow domain that serves to interact with the tail of the bacteriophage. A channel runs along the connector wide enough to allow the translocation of a double-stranded DNA molecule into the prohead. The general structure of the T3 connector is very similar to those obtained for other nonrelated bacteriophages and strongly suggests that the shape of this important viral structure is intimately related to its function.
Valpuesta
J M
JM
Centro Nacional de Biotecnología, CSIC, Madrid, 28049, Spain.
Sousa
N
N
Barthelemy
I
I
Fernández
J J
JJ
Fujisawa
H
H
Ibarra
B
B
Carrascosa
J L
JL
D002213:Capsid / Q000737:chemistry* / Q000648:ultrastructure*; D036022:Capsid Proteins; D020285:Cryoelectron Microscopy; D003460:Crystallization; D007091:Image Processing, Computer-Assisted; D008958:Models, Molecular; D020836:Protein Structure, Quaternary; D011994:Recombinant Proteins / Q000737:chemistry / Q000648:ultrastructure
D036022:Capsid Proteins; D011994:Recombinant Proteins; C073670:p8 protein, Bacteriophage T3