UNCONVENTIONAL PROCESSING OF THE 3' TERMINI OF THE EPSTEIN-BARR-VIRUS DNA-POLYMERASE MESSENGER-RNA

Northern biot analysis of the Epstein-Barr virus DNA polymerase mRNA identified two discrete sizes of virally encoded polymerase transcripts, 5.08 kb detected in strains P3HR1, Raji, W-91, and FF-41 and 3.7 kb detected solely in the prototype B95-8 strain. 3' S1-nuclease mapping and analysis of cDNA sequence generated by RNA-based PCR demonstrated that the 3.7-kb polymerase mRNA from B95-8 terminates 484 base pairs downstream of the open reading frame in a region of the genome remarkable for its lack of an apparent polyadenylylation signal. Moreover, between the cleavage point and the poly(A) tract of the cDNAs are a series of inserted nucleotides, mostly adenosine and uridine residues of unknown origin. A similar analysis of the 3' terminus of the 5.0-kb mRNA from the other cell lines revealed that polyadenylylation occurs 1.4 kb downstream of the B95-8 terminus. This region is deleted in B95-8, which accounts for the alternate upstream terminus used in B95-8. Like the 3.7-kb terminus, the 5.0-kb terminus lacks a canonical polyadenylylation signal, but contains a rarely used UAUAAA sequence 32 bp upstream of the poly(A) tail. These results indicate that the mRNA encoded by the Epstein-Barr virus DNA polymerase gene is polyadenylylated at two different termini without the use of canonical signals, raising the possibility of involvement of a virus-encoded factor in 3' processing of this message.