Defective FESTA/EAF2-mediated transcriptional activation in S-II-deficient embryonic stem cells.
S-II is a transcription stimulation factor that enhances RNA synthesis by RNA polymerase II in vitro. To elucidate the function of S-II in transcriptional activation in mammalian cells, we generated an S-II-deficient murine embryonic stem (ES) cell line, DKO20, through targeted gene disruption. The DKO20 cells were viable, grew normally, and had a stable karyotype. The ability to evoke transcriptional activation of hsp70 and c-fos genes was not significantly altered in DKO20. In contrast, transcriptional activation mediated by FESTA/EAF2, a transcription factor that interacts with S-II, was decreased in DKO20 cells. The reduced transactivation potential of FESTA/EAF2 was rescued by introducing the wild-type S-II gene in DKO20. The amino-terminal region of S-II, a binding surface for FESTA/EAF2, was essential for the recovery. These results suggest that S-II is selectively required for positive transcriptional regulation of a subset of genes in murine ES cells.
Ito
Takahiro
T
Division of Developmental Biochemistry, Graduate School of Pharmaceutical Sciences, University of Tokyo, Tokyo, Japan.
Saso
Kayoko
K
Arimitsu
Nagisa
N
Sekimizu
Kazuhisa
K
D000818:Animals; D015152:Blotting, Northern; D002460:Cell Line; D053595:Embryonic Stem Cells / Q000166:cytology / Q000378:metabolism*; D018840:HSP70 Heat-Shock Proteins / Q000235:genetics; D008156:Luciferases / Q000235:genetics / Q000378:metabolism; D051379:Mice; D009154:Mutation; D009687:Nuclear Proteins / Q000235:genetics / Q000502:physiology*; D011485:Protein Binding; D016760:Proto-Oncogene Proteins c-fos / Q000235:genetics; D012319:RNA Polymerase II / Q000172:deficiency* / Q000235:genetics; D011993:Recombinant Fusion Proteins / Q000235:genetics / Q000378:metabolism; D012045:Regulatory Sequences, Nucleic Acid / Q000235:genetics; D015534:Trans-Activators / Q000235:genetics / Q000502:physiology*; D015533:Transcriptional Activation; D014162:Transfection; D020798:Two-Hybrid System Techniques
C483108:FESTA protein, mouse; D018840:HSP70 Heat-Shock Proteins; D009687:Nuclear Proteins; D016760:Proto-Oncogene Proteins c-fos; D011993:Recombinant Fusion Proteins; D015534:Trans-Activators; D008156:Luciferases; D012319:RNA Polymerase II