Organization of splicing factors in adenovirus-infected cells reflects changes in gene expression during the early to late phase transition

Abstract

The spatial distribution of splicing factors is temporally regulated during adenovirus (ad) infection. Here we focus on two splicing factor distribution patterns characteristic of ad-infected cells. During the intermediate phase splicing factors surround sites of viral DNA accumulation in regions of high transcriptional activity. This distribution appears as a series of interconnected rings when viewed by microscopy. We refer to cells with this staining pattern as "ring cells." We have previously shown that at late times after infection, splicing factors are present in discrete structures identified as enlarged interchromatin granules (IGs) that also contain spliced viral RNA. We refer to cells with this pattern as "cluster cells." We determined which steps in viral gene expression occurred in ring and cluster cells. We found that transcription and some splicing of viral late genes had occurred in ring cells. Late RNA was present at transcription sites in ring cells. Cluster cells contained spliced viral late RNA in nuclear IGs and in the cytoplasm. The presence of cluster cells in the infected culture was well correlated with the export of viral RNA to the cytoplasm. Cluster cells had synthesized late proteins. Our data show that the dynamic localization of splicing factors reflects changes in gene expression activity of the infected cell as it switches over to late gene expression.