Nucleotide sequence and molecular genetic analysis of the large subunit of ribonucleotide reductase encoded by vaccinia virus.

We have mapped the vaccinia virus (VV) gene encoding the large subunit of ribonucleotide reductase (VV M1) within the HindIII I restriction fragment by using an oligonucleotide probe. Nucleotide sequencing revealed a 2340-bp open reading frame (orf), 1-3, whose amino acid sequence is highly homologous to the mouse M1 protein. The 1-3 gene was expressed as an immediate-early gene product, being transcribed in a leftward direction into a 2.7-kb polyadenylated transcript. Hybrid-selected translation of cycloheximide-amplified immediate-early viral RNA demonstrated that this mRNA encoded an 86-kd protein, which agrees with the expected size of the reductase large subunit. The 5'- and 3'-boundaries of the 1-3 transcriptional unit were determined by primer extension and S1-nuclease analysis, respectively, and shown to contain sequence elements typical of other VV early genes. Surprisingly, the predicted amino acid sequence of the VV enzyme subunit shares 72.5% homology with the mouse large subunit, M1.