Adenovirus E1A proteins are closely associated with chromatin in productively infected and transformed cells.
The adenovirus E1A 243R oncoprotein encodes a potent transcription-repression function within the N-terminal 80 amino acids. Our proposed model of E1A repression predicts that E1A interacts with important cellular proteins on chromatin. Consistent with this idea, we report here that E1A proteins from in vivo formaldehyde cross-linked 293 cells are closely associated with chromatin even after several stringent purification steps including double isopycnic CsCl density gradient centrifugation and size exclusion chromatography. Likewise, E1A proteins expressed from virus during productive infection of HeLa cells are closely associated with chromatin starting at early times after infection. No other adenoviral proteins are necessary for E1A 243R protein to associate with chromatin. Analyses of chromatin from HeLa cells infected with adenovirus vectors expressing E1A 243R protein with deletions in different E1A functional domains indicate that sequences within the E1A N-terminal repression domain are needed for the majority of E1A's interactions with chromatin.
Institute for Molecular Virology, Saint Louis University School of Medicine, St. Louis, MO 63110, USA. green@slu.edu
Maurice
M
Green
Ninder K
NK
Panesar
Paul M
PM
Loewenstein
D000256:Adenoviridae / Q000235:genetics; D017500:Adenovirus E1A Proteins / Q000737:chemistry / Q000302:isolation & purification / Q000502:physiology*; D000595:Amino Acid Sequence; D002460:Cell Line; D002461:Cell Line, Transformed; D002472:Cell Transformation, Viral; D002499:Centrifugation, Density Gradient; D002500:Centrifugation, Isopycnic; D002586:Cesium / Q000737:chemistry; D002712:Chlorides / Q000737:chemistry; D002843:Chromatin / Q000737:chemistry / Q000502:physiology*; D002850:Chromatography, Gel; D002999:Clone Cells; D003432:Cross-Linking Reagents / Q000737:chemistry; D005557:Formaldehyde / Q000737:chemistry; D005822:Genetic Vectors; D006367:HeLa Cells; D006801:Humans; D007668:Kidney / Q000166:cytology; D008969:Molecular Sequence Data; D017434:Protein Structure, Tertiary; D017384:Sequence Deletion
D017500:Adenovirus E1A Proteins; D002712:Chlorides; D002843:Chromatin; D003432:Cross-Linking Reagents; D005557:Formaldehyde; D002586:Cesium; C028019:cesium chloride