Mutational analysis of the herpes simplex virus type 1 DNA packaging protein UL33.
The UL33 protein of herpes simplex virus type 1 (HSV-1) is thought to be a component of the terminase complex that mediates the cleavage and packaging of viral DNA. In this study we describe the generation and characterization of a series of 15 UL33 mutants containing insertions of five amino acids located randomly throughout the 130-residue protein. Of these mutants, seven were unable to complement the growth of the UL33-null virus dlUL33 in transient assays and also failed to support the cleavage and packaging of replicated amplicon DNA into capsids. The insertions in these mutants were clustered between residues 51 and 74 and between 104 and 116, within the most highly conserved regions of the protein. The ability of the mutants to interact with the UL28 component of the terminase was assessed in immunoprecipitation and immunofluorescence assays. All four mutants with insertions between amino acids 51 and 74 were impaired in this interaction, whereas two of the three mutants in the second region (with insertions at positions 111 and 116) were not affected. These data indicate that the ability of UL33 to interact with UL28 is probably necessary, but not sufficient, to support viral growth and DNA packaging.
Beilstein
Frauke
F
MRC Virology Unit, University of Glasgow, Church Street, Glasgow G11 5JR, United Kingdom.
Higgs
Martin R
MR
Stow
Nigel D
ND
D000595:Amino Acid Sequence; D000818:Animals; D002460:Cell Line; D006224:Cricetinae; D004252:DNA Mutational Analysis; D042003:DNA Packaging; D004279:DNA, Viral / Q000378:metabolism; D005455:Fluorescent Antibody Technique / Q000379:methods; D005816:Genetic Complementation Test / Q000379:methods; D018259:Herpesvirus 1, Human / Q000502:physiology*; D047468:Immunoprecipitation / Q000379:methods; D008969:Molecular Sequence Data; D016254:Mutagenesis, Insertional / Q000379:methods; D011485:Protein Binding; D025941:Protein Interaction Mapping; D014764:Viral Proteins / Q000235:genetics* / Q000378:metabolism*
D004279:DNA, Viral; C066670:UL33 protein, Human herpesvirus 1; D014764:Viral Proteins; C069185:DNA cleavage and packaging proteins, Herpesvirus