title: Inhibition of translation by a short element in the 5' leader of the herpes simplex virus 1 DNA polymerase transcript.
abstract: Many viruses regulate gene expression, both globally and specifically, to achieve maximal rates of replication. During herpes simplex virus 1 infection, translation of the DNA polymerase (Pol) catalytic subunit is inefficient relative to other proteins of the same temporal class (D. R. Yager, A. I. Marcy, and D. M. Coen., J. Virol. 64:2217-2225, 1990). To investigate the mechanisms involved in the inefficient translation of Pol and to determine whether this inefficient translation could affect viral replication, we performed a mutagenic analysis of the 5' end of the pol transcript. We found that a short sequence ( approximately 55 bases) in the 5' leader of the transcript is both necessary and sufficient to inhibit translation in rabbit reticulocyte lysates and sufficient to inhibit reporter gene translation in transfected cells. RNase structure mapping experiments indicated that the inhibitory element adopts a structure that contains regions of a double-stranded nature, which may interfere with ribosomal loading and/or scanning. Pol accumulated to approximately 2- to 3-fold-higher levels per mRNA in cells infected with a mutant virus containing a deletion of the approximately 55-base inhibitory element than in cells infected with a control virus containing this element. Additionally, the mutant virus replicated less efficiently than the control virus. These results suggest that the inhibitory element regulates Pol translation during infection and that its inhibition of Pol translation is beneficial for viral replication.
authors.affiliation: Dept. of Biological Chemistry and Molecular Pharmacology, Harvard Medical School, 250 Longwood Ave., Boston, MA 02115, USA.
authors.forename: Kevin F
authors.initials: KF
authors.lastname: Bryant
authors.forename: Donald M
authors.initials: DM
authors.lastname: Coen
mesh_terms: D020121:5' Untranslated Regions / Q000235:genetics / Q000502:physiology*; D000818:Animals; D002522:Chlorocebus aethiops; D004259:DNA-Directed DNA Polymerase / Q000235:genetics*; D005090:Exodeoxyribonucleases / Q000235:genetics*; D015967:Gene Expression Regulation, Viral / Q000235:genetics / Q000502:physiology; D017930:Genes, Reporter; D018259:Herpesvirus 1, Human / Q000502:physiology*; D049409:Luciferases, Firefly / Q000096:biosynthesis / Q000235:genetics; D016296:Mutagenesis; D009690:Nucleic Acid Conformation; D014176:Protein Biosynthesis; D012330:RNA, Double-Stranded; D012333:RNA, Messenger / Q000235:genetics* / Q000378:metabolism; D012367:RNA, Viral / Q000235:genetics* / Q000378:metabolism; D017384:Sequence Deletion; D014709:Vero Cells; D014764:Viral Proteins / Q000235:genetics*; D014779:Virus Replication / Q000235:genetics / Q000502:physiology
chemical_list: D020121:5' Untranslated Regions; D012330:RNA, Double-Stranded; D012333:RNA, Messenger; D012367:RNA, Viral; D014764:Viral Proteins; D049409:Luciferases, Firefly; D004259:DNA-Directed DNA Polymerase; D005090:Exodeoxyribonucleases; C043417:DNA polymerase, Simplexvirus