A mutation in helicase motif IV of herpes simplex virus type 1 UL5 that results in reduced growth in vitro and lower virulence in a murine infection model is related to the predicted helicase structure.
A variant was selected from a clinical isolate of herpes simplex virus type 1 (HSV-1) during a single passage in the presence of a helicase-primase inhibitor (HPI) at eight times the IC(50). The variant was approximately 40-fold resistant to the HPI BAY 57-1293 and it showed significantly reduced growth in tissue culture with a concomitant reduction in virulence in a murine infection model. The variant contained a single mutation (Asn342Lys) in the UL5 predicted functional helicase motif IV. The Asn342Lys mutation was transferred to a laboratory strain, PDK cl-1, and the recombinant acquired the expected resistance and reduced growth characteristics. Comparative modelling and docking studies predicted the Asn342 position to be physically distant from the HPI interaction pocket formed by UL5 and UL52 (primase). We suggest that this mutation results in steric/allosteric modification of the HPI-binding pocket, conferring an indirect resistance to the HPI. Slower growth and moderately reduced virulence suggest that this mutation might also interfere with the helicase-primase activity.
Biswas
Subhajit
S
Department of Veterinary Medicine, University of Cambridge, Cambridge, UK.
Miguel
Ricardo Núñez
RN
Department of Biochemistry, University of Cambridge, Cambridge, UK.
Sukla
Soumi
S
Department of Veterinary Medicine, University of Cambridge, Cambridge, UK.
Field
Hugh J
HJ
Department of Veterinary Medicine, University of Cambridge, Cambridge, UK.
D019943:Amino Acid Substitution / Q000235:genetics; D000818:Animals; D000998:Antiviral Agents / Q000494:pharmacology*; D004265:DNA Helicases / Q000737:chemistry / Q000235:genetics / Q000502:physiology*; D019915:DNA Primase / Q000737:chemistry / Q000235:genetics / Q000502:physiology*; D024882:Drug Resistance, Viral; D018259:Herpesvirus 1, Human / Q000235:genetics / Q000254:growth & development / Q000472:pathogenicity*; D051379:Mice; D008958:Models, Molecular; D020125:Mutation, Missense; D011725:Pyridines / Q000494:pharmacology*; D013449:Sulfonamides; D013844:Thiazoles / Q000494:pharmacology*; D014764:Viral Proteins / Q000737:chemistry / Q000235:genetics / Q000502:physiology*; D014774:Virulence; D037521:Virulence Factors / Q000737:chemistry / Q000235:genetics / Q000502:physiology*
D000998:Antiviral Agents; D011725:Pyridines; D013449:Sulfonamides; D013844:Thiazoles; D014764:Viral Proteins; D037521:Virulence Factors; C453221:pritelivir; D019915:DNA Primase; C062021:helicase-primase, Human herpesvirus 1; D004265:DNA Helicases