Expression of the mutagenic peptide of herpes simplex virus type 1 in virus-infected cells.
A fragment of DNA from within the minimum transforming region (mtr-1) of herpes simplex virus type 1 (HSV-1) is known to raise the mutation frequency of cells. This activity has been attributed to a viral protein whose properties are largely unknown. Antiserum was raised to a synthetic peptide of a predicted amino acid sequence from the protein, and was found to react with cells that were infected by HSV-1 in an ELISA and by immunocytochemical staining. A combination of immunoprecipitation and immunoblotting techniques confirmed that the epitope is located at the carboxy terminus of the UL26 gene product and is downstream of epitopes that are recognized by two monoclonal antibodies. The mutagenic peptide was different from the conventional gene product of UL26 in that: (a) It was expressed from a different reading frame, (b) It was expressed earlier in infection, and (c) It bound DNA, and thus could be separated by DNA-cellulose chromatography. An RT-PCR experiment revealed two deletions in the cDNA, suggesting that RNA splicing could account for the frameshift. Examination of the DNA sequence of the region also revealed a potential ribosomal frame-shift site. The mutagenic peptide of HSV-1 is therefore a product of the UL26 gene which is expressed with a different carboxy terminus early in infection, and this could be due either to RNA splicing or to ribosomal frame-shifting.
Das
C M
CM
Department of Microbiology, University of Texas Dental Branch, Houston 77225.
Zhang
S
S
Shillitoe
E J
EJ
D000595:Amino Acid Sequence; D000818:Animals; D000911:Antibodies, Monoclonal; D001483:Base Sequence; D002472:Cell Transformation, Viral; D002522:Chlorocebus aethiops; D002846:Chromatography, Affinity; D017931:DNA Primers; D004279:DNA, Viral / Q000378:metabolism; D004797:Enzyme-Linked Immunosorbent Assay; D016679:Genome, Viral; D018259:Herpesvirus 1, Human / Q000235:genetics / Q000378:metabolism*; D015151:Immunoblotting; D007150:Immunohistochemistry; D008969:Molecular Sequence Data; D016296:Mutagenesis; D016133:Polymerase Chain Reaction; D016364:Reading Frames; D012697:Serine Endopeptidases / Q000032:analysis / Q000096:biosynthesis* / Q000302:isolation & purification; D014709:Vero Cells; D014764:Viral Proteins
D000911:Antibodies, Monoclonal; D017931:DNA Primers; D004279:DNA, Viral; D014764:Viral Proteins; C079287:scaffold protein, Herpes simplex virus-1; D012697:Serine Endopeptidases