DNA-dependent protein kinase is a novel interaction partner for Ets-1 isoforms.
The Ets-1 transcription factor plays an important role in various physiological and pathological processes. These diverse roles of Ets-1 are likely to depend on its interaction partner proteins. We used our previously developed, recombinant biotinylated Ets-1 that conserves native Ets-1 properties to identify new interaction partners. Here, based on results from streptavidin pull-down assays, mass spectrometry and co-immunoprecipitation, we report a novel interaction partner for Ets-1 isoforms: a heterotrimeric complex of DNA-dependent protein kinase (DNA-PK), made up of Ku70, Ku86, and DNA-PK catalytic subunit (DNA-PKcs). Kinase assays performed in vitro showed that DNA-PK phosphorylates the Ets-1 protein. Furthermore, we demonstrated that Ku86, but not Ku70 or DNA-PKcs, down-regulated the transcriptional activity of Ets-1 when analysed using a reporter gene assay. These results illustrate how detecting novel molecular interactions may provide new clues for understanding the diverse functions of Ets-1.
Choul-li
Souhaila
S
CNRS UMR 8161, Institut de Biologie de Lille, Institut Pasteur de Lille, Univ Lille Nord de France, IFR 142, BP 447, 1 rue du Pr Calmette, 59021 Lille Cedex, France.
Drobecq
Hervé
H
Aumercier
Marc
M
D034961:Antigens, Nuclear / Q000378:metabolism*; D051747:DNA-Activated Protein Kinase / Q000378:metabolism*; D004268:DNA-Binding Proteins / Q000378:metabolism*; D015536:Down-Regulation; D017930:Genes, Reporter; D006801:Humans; D047468:Immunoprecipitation; D000072200:Ku Autoantigen; D010766:Phosphorylation; D020033:Protein Isoforms / Q000378:metabolism; D050784:Proto-Oncogene Protein c-ets-1 / Q000378:metabolism*; D014158:Transcription, Genetic
D034961:Antigens, Nuclear; D004268:DNA-Binding Proteins; C494041:ETS1 protein, human; D020033:Protein Isoforms; D050784:Proto-Oncogene Protein c-ets-1; D051747:DNA-Activated Protein Kinase; C000606509:Xrcc6 protein, human; D000072200:Ku Autoantigen