Evidence that two latency-associated transcripts of herpes simplex virus type 1 are nonlinear

The latency-associated transcripts (LATs) of herpes simplex virus type 1 (HSV-1) are the only viral gene products that accumulate to abundant levels in latently infected cells. Others have reported species of 2.0, 1.50, and 1.45 kb; only the 2.0-kb species is seen in productively infected cells, and there is evidence that it behaves as an intron. We examined the LATs both in trigeminal ganglia of latently infected mice and in productively infected cultures of monkey CV-1 cells. After glyoxalation, RNA was subjected to high-resolution agarose gel electrophoresis and Northern (RNA) analysis, a procedure capable of resolving linear and nonlinear RNA species. Under these conditions, we resolved the 2.0-kb LAT into two species; the slower species was much more abundant and had a mobility significantly slower than expected for a linear RNA. To test the hypothesis that this RNA was in fact nonlinear, we used partial hydrolysis by sodium carbonate and oligonucleotide-directed RNase H digestion. These procedures changed the mobility of the slower species into that of the faster species. Similarly, the mobility of the 1.50-kb LAT, which was much more abundant than the 1.45-kb LAT, was changed by these procedures to that of the 1.45-kb LAT. Our data show that the two major LAT species are nonlinear, and they support an interpretation of stable lariat structures.