Resistance of herpes simplex virus type 1 against different phosphonylmethoxyalkyl derivatives of purines and pyrimidines due to specific mutations in the viral DNA polymerase gene

Drug-resistant strains of herpes simplex virus type 1 (HSV-1) were selected under the pressure of (S)-3-hydroxy-2-phosphonylmethoxypropyl (HPMP) deriv atives of cytosine (HPMPC, cidofovir) and adenine (HPMPA) and 2-phosphonylm ethoxyethyl (PME) derivatives of adenine (PMEA, adefovir) and 2,6-diaminopu rine (PMEDAP). HPMPC-resistant (HPMPCr) and HPMPA(r) strains were cross-res istant to one another, but they remained sensitive to foscarnet (PFA), acyc lovir (ACV) and the PME derivatives, while the PMEA(r) and PMEDAP(r) strain s showed cross-resistance to PFA and ACV. The PMEA(r), PMEDAP(r) and PFA(r) mutants all revealed a single nucleotide change resulting in a Ser-724 to Asn mutation with in the conserved region II of the DNA polymerase. Two HPM PA(r) clones and one HPMPCr clone possessed single amino acid changes in th e DNA polymerase (HPMPA(r) clone D1, Leu-1007 to Met; HPMPA(r) clone B5, Il e-1028 to Thr; HPMPCr clone C3, Val-573 to Met). The HPMPCr clone A4 contai ned two mutations, Ala-136 to Thr and Arg-700 to Met. The mutation at posit ion 136, located outside the catalytic domain of the enzyme, was not detect ed in other HPMPCr clones, suggesting that this mutation may not be respons ible for the resistant phenotype. Residue 573 is located within the 3' --> 5' exonuclease editing domain close to the catalytically important residues Tyr-577 and Asp-581. Similarly, residue 700 is located in the palm subdoma in of the catalytic domain, adjacent to the Asp residues 717, 886 and 888 t hat are vital for polymerase activity. The HPMPA(r) mutations at residues 1 007 and 1028, beyond the last conserved region, still fall within the thumb subdomain of the catalytic domain. The different drug-resistant mutants va ried in neurovirulent behaviour, the HPMPCr strains showing reduced neurovi rulence compared with the wild-type.