title: Assembly of bacteriophage P2 capsids from capsid protein fused   to internal scaffolding protein.
abstract: Most tailed bacteriophages with double-stranded DNA genomes code for a scaffolding protein, which is required for capsid assembly, but is removed during capsid maturation and DNA packaging. The gpO scaffolding protein of bacteriophage P2 also doubles as a maturation protease, while the scaffolding activity is confined to a 90 residue C-terminal "scaffolding" domain. Bacteriophage HK97 lacks a separate scaffolding protein; instead, an N-terminal "delta" domain in the capsid protein appears to serve an analogous role. We asked whether the C-terminal scaffolding domain of gpO could work as a delta domain when fused to the gpN capsid protein. Varying lengths of C-terminal sequences from gpO were fused to the N-terminus of gpN and expressed in E. coli. The presence of just the 41 C-terminal residues of gpO increased the fidelity of assembly and promoted the formation of closed shells, but the shells formed were predominantly small, 40 nm shells, compared to the normal, 55 nm P2 procapsid shells. Larger scaffolding domains fused to gpN caused the formation of shells of varying size and shape. The results suggest that while fusing the scaffolding protein to the capsid protein assists in shell closure, it also restricts the conformational variability of the capsid protein.
authors.affiliation: Department of Microbiology, University of Alabama at Birmingham, AL 35294, USA
authors.forename: Jenny R
authors.initials: JR
authors.lastname: Chang
authors.forename: Michael S
authors.initials: MS
authors.lastname: Spilman
authors.forename: Terje
authors.initials: T
authors.lastname: Dokland
mesh_terms: D017901:Bacteriophage P2 / Q000235:genetics / Q000502:physiology*; D036022:Capsid Proteins / Q000235:genetics / Q000378:metabolism*; D020285:Cryoelectron Microscopy; D004926:Escherichia coli / Q000821:virology; D011993:Recombinant Fusion Proteins / Q000235:genetics / Q000378:metabolism; D014764:Viral Proteins / Q000235:genetics / Q000378:metabolism*; D014771:Virion / Q000378:metabolism / Q000648:ultrastructure; D019065:Virus Assembly
chemical_list: D036022:Capsid Proteins; D011993:Recombinant Fusion Proteins; D014764:Viral Proteins