Interaction of YY1 with E2Fs, mediated by RYBP, provides a mechanism for specificity of E2F function.
To explore mechanisms for specificity of function within the family of E2F transcription factors, we have identified proteins that interact with individual E2F proteins. A two-hybrid screen identified RYBP (Ring1- and YY1-binding protein) as a protein that interacts specifically with the E2F2 and E2F3 family members, dependent on the marked box domain in these proteins. The Cdc6 promoter contains adjacent E2F- and YY1-binding sites, and both are required for promoter activity. In addition, YY1 and RYBP, in combination with either E2F2 or E2F3, can stimulate Cdc6 promoter activity synergistically, dependent on the marked box domain of E2F3. Using chromatin immunoprecipitation assays, we show that both E2F2 and E2F3, as well as YY1 and RYBP, associate with the Cdc6 promoter at G(1)/S of the cell cycle. In contrast, we detect no interaction of E2F1 with the Cdc6 promoter. We suggest that the ability of RYBP to mediate an interaction between E2F2 or E2F3 and YY1 is an important component of Cdc6 activation and provides a basis for specificity of E2F function.
Schlisio
Susanne
S
Department of Molecular Genetics and Microbiology and Howard Hughes Medical Institute, Duke University Medical Center, Durham, NC 27710, USA.
Halperin
Terri
T
Vidal
Miguel
M
Nevins
Joseph R
JR
D016475:3T3 Cells; D000818:Animals; D018797:Cell Cycle Proteins / Q000235:genetics; D002843:Chromatin / Q000378:metabolism; D004268:DNA-Binding Proteins; D050684:E2F Transcription Factors; D050687:E2F1 Transcription Factor; D050703:E2F2 Transcription Factor; D050699:E2F3 Transcription Factor; D050981:Erythroid-Specific DNA-Binding Factors; D006801:Humans; D051379:Mice; D011233:Precipitin Tests; D011401:Promoter Regions, Genetic; D011485:Protein Binding; D012097:Repressor Proteins / Q000378:metabolism*; D029701:Saccharomyces cerevisiae Proteins; D014157:Transcription Factors / Q000378:metabolism*; D014407:Tumor Cells, Cultured; D050986:YY1 Transcription Factor
C066503:CDC6 protein, S cerevisiae; D018797:Cell Cycle Proteins; D002843:Chromatin; D004268:DNA-Binding Proteins; D050684:E2F Transcription Factors; D050687:E2F1 Transcription Factor; C493753:E2F1 protein, human; D050703:E2F2 Transcription Factor; C493761:E2F2 protein, human; D050699:E2F3 Transcription Factor; C493754:E2f1 protein, mouse; D050981:Erythroid-Specific DNA-Binding Factors; D012097:Repressor Proteins; C120616:Rybp protein, mouse; D029701:Saccharomyces cerevisiae Proteins; D014157:Transcription Factors; D050986:YY1 Transcription Factor; C494698:YY1 protein, human; C494699:Yy1 protein, mouse