EVIDENCE THAT 2 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 abundan t 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, W e examined the LATs both in trigeminal ganglia of latently infected mi ce and in productively infected cultures of monkey CV-I cells, After g lyoxalation, RNA was subjected to high-resolution agarose gel electrop horesis 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 abun dant and had a mobility significantly slower than expected for a linea r RNA. To test the hypothesis that this RNA was in fact nonlinear, we used partial hydrolysis by sodium carbonate and oligonucleotide-direct ed RNase H digestion, These procedures changed the mobility of the slo wer species into that of the faster species, Similarly, the mobility o f 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 s how that the two major LAT species are nonlinear, and they support an interpretation of stable lariat structures.