The essential and nonessential transcription units for viral protein synthesis and DNA replication of porcine circovirus type 2.

During porcine circovirus (PCV) replication in PK15 cells, nine PCV type 2 (PCV2)-specific RNAs are synthesized. They include the capsid RNA (CR), five Rep-associated RNAs (Rep, Rep', Rep3a, Rep3b, and Rep3c), and three NS-associated RNAs (NS515, NS672, and NS0). In this work, mutational analyses were conducted to investigate the involvement of each PCV2 transcription unit in viral protein synthesis and DNA replication. The results demonstrated that a stop codon introduced at the very 5'-end of CR did not affect Rep-associated antigens or viral DNA synthesis. Altering the consensus dinucleotides at the splice junctions of the minor RNAs (Rep3a, Rep3b, Rep3c, NS515, and NS672) or introducing a stop codon in the abundant NS0 RNA also did not have any effect on viral protein synthesis or DNA replication. However, mutations that resulted in truncated Rep or Rep' proteins caused greater than 99% reduction of viral protein synthesis and complete shut down of viral DNA replication. These results demonstrated that both Rep and Rep' are absolutely essential for PCV2 replication.