TitleLocalization of the antigenic sites and intrinsic protein kinase domain within a 300 amino acid segment of the ribonucleotide reductase large subunit from herpes simplex virus type 2.
Publication TypeJournal Article
Year of Publication1992
AuthorsAli MA, Prakash SS, Jariwalla RJ
Date Published1992 Mar
KeywordsBase Sequence, Cloning, Molecular, Epitopes, Microscopy, Fluorescence, Molecular Sequence Data, Mutagenesis, Site-Directed, Protein Kinases, Restriction Mapping, Ribonucleotide Reductases, Simplexvirus

The 140-kDa ribonucleotide reductase (RR1) protein of herpes simplex virus type 2 (HSV-2) functions as the large subunit of virus-specified RR1 and exhibits an intrinsic protein kinase (PK) activity at its unique NH2-terminal region. The N-terminal half of RR1 contains the protein and DNA functions of the morphological transforming region III (mtrIII) of HSV-2. In the present study, we have expressed a number of truncated RR1 derivatives in a mammalian expression vector containing NH2-terminal RR1 gene fragments and amber mutants generated by site-specific mutagenesis. These derivatives, synthesized in transient expression assays, were used as test antigens to localize the epitopes of a panel of HSV-2 RR1-reactive monoclonal antibodies and to fine-map the PK catalytic domain. Our data show that the epitope for HSV-2-specific monoclonal antibody 6A-6 is located in a region of RR1 protein spanning aa 72-350. The epitopes for cross-reactive antibodies to HSV RR1, i.e., 48S and 51S, are formed predominantly by a stretch of amino acid residues specified by aa 350-376 of the RR1 molecule. The 6A-6 antibody utilized in conjunction with the RR1 amber mutants has allowed us to define a 278 aa domain within the NH2-terminal half of the 140-kDa RR1 (aa 72-350) that is sufficient for PK activity.

Alternate JournalVirology
PubMed ID1371028
Grant ListCA 42467 / CA / NCI NIH HHS / United States