title: Cryo-reconstructions of P22 polyheads suggest that phage assembly is nucleated by trimeric interactions among coat proteins.
abstract: Bacteriophage P22 forms an isometric capsid during normal assembly, yet when the coat protein (CP) is altered at a single site, helical structures (polyheads) also form. The structures of three distinct polyheads obtained from F170L and F170A variants were determined by cryo-reconstruction methods. An understanding of the structures of aberrant assemblies such as polyheads helps to explain how amino acid substitutions affect the CP, and these results can now be put into the context of CP pseudo-atomic models. F170L CP forms two types of polyhead and each has the CP organized as hexons (oligomers of six CPs). These hexons have a skewed structure similar to that in procapsids (precursor capsids formed prior to dsDNA packaging), yet their organization differs completely in polyheads and procapsids. F170A CP forms only one type of polyhead, and though this has hexons organized similarly to hexons in F170L polyheads, the hexons are isometric structures like those found in mature virions. The hexon organization in all three polyheads suggests that nucleation of procapsid assembly occurs via a trimer of CP monomers, and this drives formation of a T = 7, isometric particle. These variants also form procapsids, but they mature quite differently: F170A expands spontaneously at room temperature, whereas F170L requires more energy. The P22 CP structure along with scaffolding protein interactions appear to dictate curvature and geometry in assembled structures and residue 170 significantly influences both assembly and maturation.
authors.affiliation: Department of Chemistry & Biochemistry, University of California, San Diego, La Jolla, CA, USA.
authors.forename: Kristin N
authors.initials: KN
authors.lastname: Parent
authors.forename: Robert S
authors.initials: RS
authors.lastname: Sinkovits
authors.forename: Margaret M
authors.initials: MM
authors.lastname: Suhanovsky
authors.forename: Carolyn M
authors.initials: CM
authors.lastname: Teschke
authors.forename: Edward H
authors.initials: EH
authors.lastname: Egelman
authors.forename: Timothy S
authors.initials: TS
authors.lastname: Baker
mesh_terms: D017100:Bacteriophage P22 / Q000378:metabolism / Q000502:physiology* / Q000648:ultrastructure; D002213:Capsid; D036022:Capsid Proteins / Q000737:chemistry / Q000378:metabolism*; D046529:Microscopy, Electron, Transmission; D008958:Models, Molecular; D011485:Protein Binding; D019065:Virus Assembly
chemical_list: D036022:Capsid Proteins