N. Prang et al., Latency of Epstein-Barr virus is stabilized by antisense-mediated control of the viral immediate-early gene BZLF-1, J MED VIROL, 59(4), 1999, pp. 512-519
The ability of the Epstein-Barr virus (EBV) to avoid lytic replication and
to establish a latent infection in B-lymphocytes is fundamental for its lif
elong persistence and the pathogenesis of various EBV-associated diseases.
The viral immediate-early gene BZLF-1 plays a key role for the induction of
lytic replication and its activity is strictly regulated on different leve
ls of gene expression. Recently, it was demonstrated that BZLF-1 is also co
ntrolled by a posttranscriptional mechanism. Transient synthesis of a mutat
ed competitor RNA saturated this mechanism and caused both expression of th
e BZLF-1 protein and the induction of lytic viral replication. Using short
overlapping fragments of the competitor, it is shown that this control acts
on the unspliced primary transcript. RT-PCR demonstrated unspliced BZLF-1
RNA in latently infected B-lymphocytes in the absence of BZLF-1 protein. Du
e to the complementarity of the gene BZLF-1 and the latency-associated gene
EBNA-1 on the opposite strand of the genome, we propose an antisense-media
ted mechanism. RNase protection assays demonstrated transcripts in antisens
e orientation to the BZLF-1 transcript during latency, which comprise a com
parable constellation to other herpesviruses. A combined RNAse protection/R
T-PCR assay detected the double-stranded hybrid RNA, consisting of the unsp
liced BZLF-1 transcript and a noncoding intron of the EBNA-1 gene. Binding
of BZLF-1 transcripts is suggested to be an important backup control mechan
ism in addition to transcriptional regulation, stabilizing latency and prev
enting inappropriate lytic viral replication in vivo. (C) 1999 Wiley-Liss,
Inc.