Degradation of the retinoblastoma tumor suppressor by the human papillomavirus type 16 E7 oncoprotein is important for functional inactivation and is separable from proteasomal degradation of E7.
The steady-state level and metabolic half-life of retinoblastoma tumor suppressor protein pRB are decreased in cells that express high-risk human papillomavirus (HPV) E7 proteins. Here we show that pRB degradation is a direct activity of E7 and does not reflect a property of cell lines acquired during the selection process for E7 expression. An amino-terminal domain of E7 that does not directly contribute to pRB binding but is required for transformation is also necessary for E7-mediated pRB degradation. Treatment with inhibitors of the 26S proteasome not only blocks E7-mediated pRB degradation but also causes the stabilization of E7. Mutagenic analyses, however, reveal that the processes of proteasomal degradation of E7 and pRB are not linked processes. HPV type 16 E7 also targets the pRB-related proteins p107 and p130 for destabilization by a proteasome-dependent mechanism. Using the SAOS2 flat-cell assay as a biological indicator for pRB function, we demonstrate that pRB degradation, not solely binding, is important for the E7-induced inactivation of pRB.
Gonzalez
S L
SL
Program in Biological Sciences in Public Health, Harvard School of Public Health, Harvard Medical School, Boston, Massachusetts 02115, USA.
Stremlau
M
M
He
X
X
Basile
J R
JR
Münger
K
K
D003546:Cysteine Endopeptidases / Q000378:metabolism; D015967:Gene Expression Regulation, Viral; D006801:Humans; D009097:Multienzyme Complexes / Q000378:metabolism; D009856:Oncogene Proteins, Viral / Q000235:genetics* / Q000378:metabolism; D027383:Papillomaviridae / Q000502:physiology*; D050725:Papillomavirus E7 Proteins; D030361:Papillomavirus Infections / Q000235:genetics / Q000378:metabolism / Q000821:virology*; D046988:Proteasome Endopeptidase Complex; D016160:Retinoblastoma Protein / Q000235:genetics* / Q000378:metabolism; D014407:Tumor Cells, Cultured; D014412:Tumor Virus Infections / Q000235:genetics / Q000378:metabolism / Q000821:virology*; D014779:Virus Replication / Q000235:genetics
D009097:Multienzyme Complexes; D009856:Oncogene Proteins, Viral; D050725:Papillomavirus E7 Proteins; D016160:Retinoblastoma Protein; C059731:oncogene protein E7, Human papillomavirus type 16; D003546:Cysteine Endopeptidases; D046988:Proteasome Endopeptidase Complex