In vitro assembly of bacteriophage P4 procapsids from purified capsid and scaffolding proteins.
Bacteriophage P4 is a satellite virus of bacteriophage P2, which has acquired the ability to utilize the structural gene products of P2 to assemble its own capsid. The normal P2 capsid has a T = 7 icosahedral structure comprised of the gpN-derived capsid protein, whereas the capsid produced under the control of P4 has a smaller, T = 4 structure. The protein responsible for this size determination is the P4-coded gene product Sid, which forms an external scaffold on the P4 procapsid. Using an in vitro assembly system, we show that gpN and Sid can coassemble into procapsid-like particles, indistinguishable from those produced in vivo, in the absence of any other gene products. The fidelity of the assembly reaction is enhanced by the inclusion of PEG and has a pH optimum between 8.0 and 8.5. Analysis of the assembly properties of truncated versions of Sid and gpN suggests that the amino-terminal part of Sid is involved in gpN binding, while the carboxyl-terminal part forms trimeric Sid-Sid interactions, and that the first 31 amino acids of gpN are required for binding to Sid as well as for size determination.
Wang
S
S
Institute of Molecular Agrobiology, The National University off Singapore, 117604 Singapore.
Palasingam
P
P
Nøkling
R H
RH
Lindqvist
B H
BH
Dokland
T
T
D001435:Bacteriophages / Q000737:chemistry / Q000235:genetics / Q000378:metabolism* / Q000648:ultrastructure; D002213:Capsid / Q000235:genetics / Q000302:isolation & purification* / Q000378:metabolism* / Q000648:ultrastructure; D036022:Capsid Proteins; D002474:Cell-Free System; D002499:Centrifugation, Density Gradient; D006023:Glycoproteins / Q000235:genetics / Q000302:isolation & purification / Q000378:metabolism / Q000648:ultrastructure; D008027:Light; D008854:Microscopy, Electron; D008954:Models, Biological; D008958:Models, Molecular; D011485:Protein Binding; D020836:Protein Structure, Quaternary; D012542:Scattering, Radiation; D017384:Sequence Deletion / Q000235:genetics; D015678:Viral Structural Proteins / Q000235:genetics / Q000302:isolation & purification* / Q000378:metabolism* / Q000648:ultrastructure; D019065:Virus Assembly
D036022:Capsid Proteins; D006023:Glycoproteins; C095129:Sid protein, Bacteriophage P4; D015678:Viral Structural Proteins