Increased levels of E2F-1-dependent DNA binding activity after UV- or gamma-irradiation.
In mammalian cells, DNA damage induces robust changes in gene expression and these changes contribute to the proper execution of cellular responses to DNA damage, including DNA repair, cell cycle arrest and apoptosis. The transcription factor E2F-1 has been suggested to play a key role in the regulation of cell cycle-dependent gene expression and apoptosis. These activities depend on the ability of E2F-1 to form functionally active DNA binding complexes. Here we describe an assay that allows one to measure E2F-1 DNA binding activity in naive cells. We find that DNA damage, generated by UV- or gamma-irradiation, prompts increased production of E2F-1 DNA binding activity, which, at least in part, originates from alterations in E2F-1 protein levels. These findings represent an indication for a role of the transcription factor E2F-1 in the DNA damage response pathway.
Höfferer
M
M
Friedrich Miescher Institut, Maulbeerstrasse 66, CH-4058 Basel, Switzerland and Departement Forschung, Kantonsspital Basel, Hebelstrasse 20, CH-4031 Basel, Switzerland.
Wirbelauer
C
C
Humar
B
B
Krek
W
W
D000595:Amino Acid Sequence; D002352:Carrier Proteins; D018797:Cell Cycle Proteins / Q000378:metabolism; D004249:DNA Damage; D004268:DNA-Binding Proteins / Q000378:metabolism*; D050684:E2F Transcription Factors; D050687:E2F1 Transcription Factor; D005720:Gamma Rays; D006801:Humans; D008969:Molecular Sequence Data; D011233:Precipitin Tests; D011485:Protein Binding / Q000528:radiation effects; D056465:Retinoblastoma-Binding Protein 1; D050680:Transcription Factor DP1; D014157:Transcription Factors / Q000378:metabolism*; D014466:Ultraviolet Rays
D002352:Carrier Proteins; D018797:Cell Cycle Proteins; D004268:DNA-Binding Proteins; D050684:E2F Transcription Factors; D050687:E2F1 Transcription Factor; C493753:E2F1 protein, human; D056465:Retinoblastoma-Binding Protein 1; D050680:Transcription Factor DP1; D014157:Transcription Factors