Studies on Arginyl Transfer Ribonucleic Acid in Herpes Virus Infected Baby Hamster Kidney Cells

Abstract

SUMMARY Previous work, (Subak-Sharpe & Hay, I965, Subak-Sharpe, Shepherd & Hay, ~966), depending on the hybridization of (32P)4s RNA from herpes virus infected cells to herpes DNA and analysis of the aminoacyloligonucleotide fragments pro- duced after RNase T I digestion of (14C)- and (3H)-arginyl tRNA, suggested the presence, in infected cells of a herpes-specified arginyl tRNA. The present study indicates that these results were due to the presence of (a) radioactive RNA other than tRNA in the (32p) 4 s RNA, and (b) radioactive contaminants in the (3H)-argi- nine used to aminoacylate the tRNA. Further purification of the aminoacyl tRNA removes these (3H)-arginine contaminants. Protection of the aminoacyl tRNA ester bond by N-acetylation improved the sensitivity of the aminoacyloligonucleotide analyses and permitted tRNA-specific molecular hybridization. Using these techniques, no herpes-specified arginyl tRNA was detected in cells 7 to 9 hr post infection.