title: Differential regulation of the pocket domains of the retinoblastoma family proteins by the HPV16 E7 oncoprotein.
abstract: The human papillomavirus E7 oncoprotein binds to the retinoblastoma (Rb) tumor suppressor protein, and the binding to Rb correlates with the oncogenic potential of E7. Recent studies from several laboratories indicated that the half-life of the Rb protein is reduced in cells that are stably transformed with E7, suggesting that E7 could induce the proteolytic degradation of Rb. To investigate whether the Rb degradation is a primary effect of E7 or a result of altered cell phenotype, we sought to develop assays that can distinguish between the two possibilities. Using recombinant adenovirus expressing the human papillomavirus type 16 E7 protein, we show that the expression of E7 leads to an increased rate of decay of the Rb protein. Moreover, Rb degradation immediately follows the expression of E7 suggesting that it is an early and primary effect. Consistent with a previous study, we observed that the E7-induced degradation of Rb can be blocked by the inhibitors of the 26S proteasome. We have also developed a transient transfection assay for the E7-induced degradation of Rb. Using this assay, we show that the pocket domain of Rb is necessary and sufficient for the E7-induced degradation. However, the proteolysis is relatively specific for Rb because the level of p107 or p130 was not significantly altered by the expression of E7. Thus, although E7 binds to all three members of the Rb family of proteins, the proteolysis is much more efficient in the case of Rb. In the transient transfection assays, adenovirus E1A and SV40 large T antigen failed to induce degradation of Rb, suggesting that the Rb degradation is a unique property of the E7 oncoprotein.
authors.affiliation: Center for Molecular Biology of Oral Diseases, College of Dentistry 60612, USA.
authors.forename: E
authors.initials: E
authors.lastname: Berezutskaya
authors.forename: B
authors.initials: B
authors.lastname: Yu
authors.forename: A
authors.initials: A
authors.lastname: Morozov
authors.forename: P
authors.initials: P
authors.lastname: Raychaudhuri
authors.forename: S
authors.initials: S
authors.lastname: Bagchi
mesh_terms: D000111:Acetylcysteine / Q000031:analogs & derivatives / Q000494:pharmacology; D017500:Adenovirus E1A Proteins / Q000378:metabolism / Q000494:pharmacology; D000952:Antigens, Polyomavirus Transforming / Q000378:metabolism / Q000494:pharmacology; D002460:Cell Line; D015853:Cysteine Proteinase Inhibitors / Q000494:pharmacology; D005786:Gene Expression Regulation; D006801:Humans; D009687:Nuclear Proteins / Q000378:metabolism; D009856:Oncogene Proteins, Viral / Q000378:metabolism* / Q000494:pharmacology; D050725:Papillomavirus E7 Proteins; D010447:Peptide Hydrolases / Q000187:drug effects; D046988:Proteasome Endopeptidase Complex; D016160:Retinoblastoma Protein / Q000037:antagonists & inhibitors / Q000378:metabolism*; D050720:Retinoblastoma-Like Protein p107; D014162:Transfection
chemical_list: D017500:Adenovirus E1A Proteins; D000952:Antigens, Polyomavirus Transforming; D015853:Cysteine Proteinase Inhibitors; D009687:Nuclear Proteins; D009856:Oncogene Proteins, Viral; D050725:Papillomavirus E7 Proteins; C493844:RBL1 protein, human; D016160:Retinoblastoma Protein; D050720:Retinoblastoma-Like Protein p107; C059731:oncogene protein E7, Human papillomavirus type 16; C067713:lactacystin; D010447:Peptide Hydrolases; D046988:Proteasome Endopeptidase Complex; C061553:ATP dependent 26S protease; D000111:Acetylcysteine