Cell-specific utilization of the albumin and immunoglobulin promoters within non-replicating adenovirus genomes has been established. With such viruses plus helper viruses we now consider the impact of infection by replicating viruses on liver-specific gene expression in hepatoma cells and of the effect of replication on the previously established limits of cell-specific expression. We find that replication of any adenovirus, whether or not it contains the albumin promoter, decreases albumin and apolipoprotein A transcription in hepatoma cells but does not affect transcription of at least four other genes expressed mainly (or only) in the liver. Thus, it may be that some hepatoma-specific genes and late adenoviral genes require a factor(s) in common, and these factors become limiting during replication. In hepatoma cells, the increased copy number of the exogenous promoters resulting from viral DNA replication did not influence the cell-specific expression noted previously; only albumin and not globin or immunoglobulin promoters in the virus were active. In contrast, replication by the same viruses in HeLa cells resulted in high levels of expression from all the exogenous promoters.