Herpes Simplex Virus Type 1 DNA Cleavage Requires the Product of the UL28 Gene : Isolution and Partial Characterization of Two UL28 Insertion Mutants
Breithaupt, Amie D.
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The UL28 gene in Herpes simplex virus (Type 1) encodes the protein ICP18.5 which has been shown to be required for the cleavage and encapsidation of viral DNA (C. Addison, F. Rixon, and V. Preston, J. Gen. Virol. 71: 2377-2384, 1990; L. Tengelson, N. Pederson, P. Shaver, M. Wathen, and F. Homa, J. of Virol. 67: 3470-3480, 1993). In this study, the confirmation of two UL28 mutants, 28-214i and 28-386i, created by Linker Insertion Mutagenesis was described. The mutant, 28-214i contains 8 additional amino acids in the UL28 gene (Arg, Arg, Ser, Ala, Arg, Arg, Ser, Arg) and a BglIl restriction site at 57,517 bp. The mutant, 28-386i, contains 4 additional amino acids in the UL28 gene (Arg, Asp, Leu, Ala) as well as a BglII restriction site at 57,001 bp.' Although 28-214i and 28-386i mutants synthesize viral DNA they fail to form plaques or produce infectious virus on cell lines that do not express the UL28 gene. Southern blot analysis showed that these mutants fail to cleave concatemeric viral DNA into unit-length molecules on non-complementing cell lines. Furthermore, the two UL28 mutants may be defective in encapsidation of cleaved viral DNA.Missing pages 6, 20, 24, and 25.