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Journal of Virology, October 2002, p. 10245-10255, Vol. 76, No. 20
0022-538X/02/$04.00+0     DOI: 10.1128/JVI.76.20.10245-10255.2002
Copyright © 2002, American Society for Microbiology. All Rights Reserved.

A P22 Scaffold Protein Mutation Increases the Robustness of Head Assembly in the Presence of Excess Portal Protein

Sean D. Moore and Peter E. Prevelige, Jr.^*

Department of Microbiology, University of Alabama at Birmingham, Birmingham, Alabama 35295

Received 12 April 2002/ Accepted 8 July 2002

Bacteriophage with linear, double-stranded DNA genomes package DNA into preassembled protein shells called procapsids. Located at one vertex in the procapsid is a portal complex composed of a ring of 12 subunits of portal protein. The portal complex serves as a docking site for the DNA packaging enzymes, a conduit for the passage of DNA, and a binding site for the phage tail. An excess of the P22 portal protein alters the assembly pathway of the procapsid, giving rise to defective procapsid-like particles and aberrant heads. In the present study, we report the isolation of escape mutant phage that are able to replicate more efficiently than wild-type phage in the presence of excess portal protein. The escape mutations all mapped to the same phage genome segment spanning the portal, scaffold, coat, and open reading frame 69 genes. The mutations present in five of the escape mutants were determined by DNA sequencing. Interestingly, each mutant contained the same mutation in the scaffold gene, which changes the glycine at position 287 to glutamate. This mutation alone conferred an escape phenotype, and the heads assembled by phage harboring only this mutation had reduced levels of portal protein and exhibited increased head assembly fidelity in the presence of excess portal protein. Because this mutation resides in a region of scaffold protein necessary for coat protein binding, these findings suggest that the P22 scaffold protein may define the portal vertices in an indirect manner, possibly by regulating the fidelity of coat protein polymerization.

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* Corresponding author. Mailing address: Department of Microbiology BBRB 416/6, University of Alabama at Birmingham, 845 South 19th St., Birmingham, AL 35295. Phone: (205) 975-5327. Fax: (205) 975-5479. E-mail: prevelig{at}uab.edu.

Present address: Department of Biology, Massachusetts Institute of Technology, Cambridge, MA 02139.

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Journal of Virology, October 2002, p. 10245-10255, Vol. 76, No. 20
0022-538X/02/$04.00+0     DOI: 10.1128/JVI.76.20.10245-10255.2002
Copyright © 2002, American Society for Microbiology. All Rights Reserved.

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