Association of bovine papillomavirus E2 protein with nuclear structures in vivo.
Papillomaviruses are small DNA viruses which have the capacity to establish a persistent infection in mammalian epithelial cells. The papillomavirus E2 protein is a central coordinator of viral gene expression, genome replication, and maintenance. We have investigated the distribution of bovine papillomavirus E2 protein in nuclei of proliferating cells and found that E2 is associated with cellular chromatin. This distribution does not change during the entire cell cycle. The N-terminal transactivation domain, but not the C-terminal DNA-binding domain, of the E2 protein is responsible for this association. The majority of the full-length E2 protein can only be detected in chromatin-enriched fractions but not as a free protein in the nucleus. Limited micrococcal nuclease digestion revealed that the E2 protein partitioned to different chromatin regions. A fraction of the E2 protein was located at nuclear sites that are resistant against nuclease attack, whereas the remaining E2 resided on compact chromatin accessible to micrococcal nuclease. These data suggest that there are two pools of E2 in the cell nucleus: one that localizes on transcriptionally inactive compact chromatin and the other, which compartmentalizes to transcriptionally active nuclear structures of the cell. Our data also suggest that E2 associates with chromatin through cellular protein(s), which in turn is released from chromatin at 0.4 M salt.
Kurg
Reet
R
Institute of Technology, University of Tartu, 23 Riia Street, 51010 Tartu, Estonia. rkurg@ebc.ee
Sild
Kristiina
K
Ilves
Aigi
A
Sepp
Mari
M
Ustav
Mart
M
D000818:Animals; D016466:CHO Cells; D002453:Cell Cycle; D002467:Cell Nucleus / Q000737:chemistry* / Q000821:virology*; D002843:Chromatin / Q000737:chemistry; D006224:Cricetinae; D004261:DNA Replication; D004268:DNA-Binding Proteins / Q000032:analysis*; D010957:Plasmids; D014764:Viral Proteins / Q000032:analysis*
D002843:Chromatin; D004268:DNA-Binding Proteins; C051550:E2 protein, Bovine papillomavirus; D014764:Viral Proteins