Analysis of the 2-kilobase latency-associated transcript expressed in PC12 cells productively infected with herpes simplex virus type 1: evidence for a stable, nonlinear structure

Abstract

The major latency-associated transcript (LAT) expressed in PC12 cells productively infected with herpes simplex virus type 1 is a 2-kb, nonpolyadenylated RNA molecule that accumulates in the nuclei of infected cells. In actinomycin D-treated cells, the 2-kb LAT gene transcript has a half-life considerably greater than 12 h. After polyacrylamide gel electrophoresis, two species of the transcript were observed, a major species that was retarded in the gel and a minor species that migrated as a 1.96-kb RNA molecule. RNase H digestion after hybridization of the RNA with an oligonucleotide complementary to positions -80 to -101 relative to the 3' end of the 2-kb LAT gene transcript changed the mobility of the retarded species into that of the rapidly migrating species. Our data indicate that the 2-kb LAT gene transcript expressed in productively infected PC12 cells is present in a stable, nonlinear form.