Overexpression of Rb and E2F-1 in ataxia-telangiectasia lymphocytes.
AT cells exhibit defective cell cycle regulation following DNA damage. Previous studies have shown that induction of p53 and p21 proteins are delayed in response to ionizing radiation, resulting in the failure of G1/S checkpoint in AT cells. In this study, further investigation of the molecular mechanisms underlying G1/S phase progression in AT cells was conducted. Exponentially growing normal and AT cells were exposed to 2 Gy of ionizing radiation and the expression levels and functional activities of Rb and E2F-1 proteins were determined. We observed overexpression of hyperphosphorylated Rb and E2F-1 proteins in AT cells, which was unaffected post-irradiation. Furthermore, gel shift assays showed that E2F-1-DNA binding was constitutive in AT cells, whereas it was inhibited in control cells following exposure to ionizing radiation. The data suggests that abnormalities in the function of Rb and E2F-1 proteins may also be responsible for the failure of AT cells to arrest in the G1/S checkpoint in response to DNA damage.
Varghese
S
S
Department of Radiation Medicine, Georgetown University Medical School, Washington, DC 20007, USA.
Jung
M
M
D017502:Adenovirus E2 Proteins / Q000096:biosynthesis*; D001260:Ataxia Telangiectasia / Q000378:metabolism*; D015153:Blotting, Western; D002460:Cell Line; D004268:DNA-Binding Proteins / Q000502:physiology*; D004591:Electrophoresis, Polyacrylamide Gel; D006801:Humans; D008214:Lymphocytes / Q000378:metabolism* / Q000528:radiation effects; D010766:Phosphorylation; D010770:Phosphotransferases / Q000032:analysis; D011233:Precipitin Tests; D016160:Retinoblastoma Protein / Q000096:biosynthesis*; D015854:Up-Regulation
D017502:Adenovirus E2 Proteins; D004268:DNA-Binding Proteins; D016160:Retinoblastoma Protein; D010770:Phosphotransferases