Dynamic behavior of transcription factors on a natural promoter in living cells.
Through the use of photobleaching techniques, we examined the dynamic interaction of three members of the transcription apparatus with a target promoter in living cells. The glucocorticoid receptor (GR) interacting protein 1 (GRIP-1) exhibits a half maximal time for fluorescent recovery (tau(R)) of 5 s, reflecting the same rapid exchange as observed for GR. In contrast, the large subunit (RPB1) of RNA polymerase II (pol II) required 13 min for complete fluorescence recovery, consistent with its function as a processive enzyme. We also observe a complex induction profile for the kinetics of GR-stimulated transcription. Our results indicate that GR and GRIP-1 as components of the activating complex are in a dynamic equilibrium with the promoter, and must return to the template many times during the course of transcriptional activation.
Becker
Matthias
M
Laboratory of Receptor Biology and Gene Expression, Building 41 Room B602, Center for Cancer Research, National Cancer Institute, National Institutes of Health, Bethesda, MD 20892-5055, USA.
Baumann
Christopher
C
John
Sam
S
Walker
Dawn A
DA
Vigneron
Marc
M
McNally
James G
JG
Hager
Gordon L
GL
D000818:Animals; D002460:Cell Line; D003609:Dactinomycin / Q000494:pharmacology; D004004:Dichlororibofuranosylbenzimidazole / Q000494:pharmacology; D017404:In Situ Hybridization, Fluorescence; D066298:In Vitro Techniques; D008324:Mammary Tumor Virus, Mouse / Q000235:genetics; D051379:Mice; D051958:Nuclear Receptor Coactivator 2; D019384:Nucleic Acid Synthesis Inhibitors / Q000494:pharmacology; D011401:Promoter Regions, Genetic / Q000502:physiology*; D012319:RNA Polymerase II / Q000037:antagonists & inhibitors / Q000378:metabolism*; D011965:Receptors, Glucocorticoid / Q000378:metabolism; D014157:Transcription Factors / Q000378:metabolism*
C498002:Ncoa2 protein, mouse; D051958:Nuclear Receptor Coactivator 2; D019384:Nucleic Acid Synthesis Inhibitors; D011965:Receptors, Glucocorticoid; D014157:Transcription Factors; D003609:Dactinomycin; D004004:Dichlororibofuranosylbenzimidazole; D012319:RNA Polymerase II