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Initiation of herpes simplex virus thymidine kinase polypeptides.

TitleInitiation of herpes simplex virus thymidine kinase polypeptides.
Publication TypeJournal Article
Year of Publication1996
AuthorsEllison AR, Bishop JO
JournalNucleic Acids Res
Volume24
Issue11
Pagination2073-9
Date Published1996 Jun 1
ISSN0305-1048
KeywordsAnimals, Antiviral Agents, Arabinofuranosyluracil, Base Sequence, Bromodeoxyuridine, Codon, Electrophoresis, Polyacrylamide Gel, Ganciclovir, Gene Expression, Herpesvirus 1, Human, Male, Mice, Molecular Sequence Data, Mutagenesis, Site-Directed, Peptide Fragments, RNA, Messenger, Testis, Thymidine Kinase, Transfection
Abstract

When employed as a transgene reporter, the herpes simplex type 1 virus (HSV1) thymidine kinase gene (tk) is ectopically expressed in mouse testis. The principal testicular mRNA lacks the 5'-end of the tk reading frame. As a result the principal translation products, P2 and P3, are N-terminally truncated. These co-migrate in SDS-PAGE with polypeptides synthesised during HSV1 infection that were previously thought to be initiated at methionine codons ATG46 and ATG60. Prompted by these observations we generated modified tk genes each carrying only one of the first three ATG codons. Transfected cells expressed both full-length enzyme (P1) and P2 when only ATG1 was unmodified, P2 and P3 when only ATG46 was unmodified or P2 and a fourth polypeptide (P4) when only ATG60 was unmodified. Our observations indicate that P3 is initiated at ATG46 rather than ATG60, while P2 is initiated at a non-ATG codon rather than ATG46 and P4 is initiated at ATG60. When either of two putative non-ATG initiation codons was modified P2 was no longer produced. Cells mainly expressing either P1 or P3 exhibited the same sensitivity to Ganciclovir as cells transfected with the unaltered tk gene. P1 and P3 both have TK activity while P4 probably has none.

Alternate JournalNucleic Acids Res.
PubMed ID8668538
PubMed Central IDPMC145909