Evidence that the RNA Polymerase Usually Does Not Make a Complete Transcript of the Late Strand of Simian Virus 40 DNA

Abstract

An important model for the transcription of the late (L) strand of simian virus 40 DNA is that, late after infection of permissive monkey cells, the RNA polymerase makes a complete transcript of the L DNA strand before terminating transcription. The purpose of the current work was to test a prediction of this model, namely, that the rate of synthesis of all RNA sequences from the L DNA strand should be equal. About one-half of the L DNA strand is transcribed into late mRNA sequences and the other half into late dRNA sequences, which do not leave the nucleus. Using glucosamine to reduce the size of the intracellular UTP pool before and after a pulse-label with radioactive uridine, a pulse-chase experiment was performed to determine the half-lives of these sequences. The half-life of the late dRNA sequences was determined to be 4 min. The late mRNA sequences were degraded more slowly, on the average, than the late dRNA sequences. In a parallel experiment with similarly treated cells, it was shown that after a 2-min label with radioactive uridine there was only 0.2 times as much radioactivity in the late dRNA sequences as in the late mRNA sequences in the total cellular RNA population. The results could be combined to calculate that the rate of synthesis of the late dRNA sequences was at most 0.3 times that of the late mRNA sequences. Consequently they provide strong evidence that when the RNA polymerase transcribes the late mRNA sequences, it usually terminates transcription before all the late dRNA sequences are transcribed.