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EMBO Rep. 2002 Dec;3(12):1188-94. Epub 2002 Nov 21.

Dynamic behavior of transcription factors on a natural promoter in living cells.

Author information

1
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.

Abstract

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.

PMID:
12446572
PMCID:
PMC1308318
DOI:
10.1093/embo-reports/kvf244
[Indexed for MEDLINE]
Free PMC Article

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