Viral DNA synthesis is required for the efficient expression of specific herpes simplex virus type 1 mRNA species

Abstract

Inhibition of HSV-1 DNA synthesis with either ~abinosyladenine plus the adenosine deaminase inhibitor pentostatin, or with arabinosylthymine, showed a viral mRNA population identical to that seen prior to viral DNA replication (early) by the criteria of quantitative hybridization, specific mRNA species identifiable by Southern blot hybridization of size-~aetionated RNA, and migration of polypeptides resolved by in vitro translation of purified viral mRNA. The amount of viral mRNA associated with infected cell polyribosomes was determined by quantitative DNA excess solution hybridization. At 2 hr postinfection (p.i.) (before viral DNA synthesis) and in drug-treated cells at 6 hr p.i., the majority of the polyadenylated RNA was cell specific with some virus-specific RNA detectable. In contrast, at 6 hr p.i., in the absence of drugs (during m~imum viral DNA synthesis), nearly all the polyadenylated polyribosomal RNA was viral. Blot hybridization of size-fractionated viral RNA confirmed several specific differences between the viral mRNA species occurring before and after HSV-1 DNA synthesis, which have been reported previously from this laboratory. These differences also were reflected in the in &ro translation products encoded by the viral mRNAs. The mRNA species and the eneoded polypeptides that were present in the absence of viral DNA synthesis are a subset of those viral mRNA species and polypeptides expressed in the presence of viral DNA synthesis. The viral mRNA species fall into several groups based on their relative abundance at various times of infection. These data suggest that, in the normal virus infection cycle, the onset of viral DNA synthesis is necessary for normal expression of later viral genes.

Identification of a Herpes Simplex Virus Type 1 Polypeptide Which Is a Component of the Virus-induced Ribonucleotide Reductase

“…(Vmw136; ICP6) which is an essential component of the virus-induced enzyme. The mRNA of Vmwl36 is a major early species although small amounts can be detected in virus-infected cells when cycloheximide has been added from the time of virus infection (Holland et aL, 1979(Holland et aL, , 1980Watson et at., 1979;McLauchlan & Clements, 1982). The polypeptide Vmwl36 is phosphorylated (Marsden et al, 1978;Wilcox et al, 1980) and is predominantly cytoplasmic in its location (Wilcox et al, 1980;Showalter et al, 1981).…”

HSV trafficking and development of gene therapy vectors with applications in the nervous system

“…The late genes, which encode structural proteins, are transactivated by the IE proteins after the onset of DNA replication. [84][85][86] The assembly of HSV particles occurs in the nucleus where capsids attach to modified patches of the inner lamella of the nuclear membrane. Budding of DNAcontaining capsids occurs at these sites, where enveloped particles are formed.…”

Intranuclear Localization of Herpes Simplex Virus Immediate-Early and Delayed-Early Proteins: Evidence that ICP 4 Is Associated with Progeny Virus DNA

“…Weissbach et al, 1973;Purifoy & Powell, 1976;Hoffman & Cheng, 1978;Banks et al, 1983;Kit & Dubbs, 1963;Dutia, 1983). Normal rates of late or y-protein synthesis are dependent on the successful replication of virus DNA (Powell et al, 1975;Honess & Watson, 1977;Holland et al, 1980).…”