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EMBO reports 3, 12, 1188–1194 (2002)
doi:10.1093/embo-reports/kvf244
Published online: December 2002
Dynamic behavior of transcription factors on a natural promoter in living cells
Matthias Becker1, Christopher Baumann1, Sam John1, Dawn A. Walker1, Marc Vigneron2, James G. McNally1 & Gordon L. Hager1
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1 Laboratory of Receptor Biology and Gene Expression, Building
41 Room B602, Center for Cancer Research, National Cancer Institute,
NIH, Bethesda, MD 20892-5055, USA
2 CR1 INSERM, UMR 7100 CNRS-ULP E.S.B.S., 1 bld Sébastien Brant, 67400 Illkirch, France
To whom correspondence should be addressed
Gordon L. Hager Tel: +1 301 496 9867; Fax: +1 301 496 4951; hagerg@exchange.nih.gov
Received 26 June 2002; Accepted 30 October 2002.
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Abstract
Through the use of photobleaching techniques, we examined the dynamic interaction of three members of the transcrsiption 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 ( 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.
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