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Letter
Nature 442, 1054-1057 (31 August 2006) | doi:10.1038/nature05067; Received 23 April 2006; Accepted 10 July 2006; Published online 23 August 2006
There is a Brief Communication Arising (8 October 2009) associated with this document.
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Proteolytic turnover of the Gal4 transcription factor is not required for function in vivo
Kip Nalley1, Stephen Albert Johnston1,2 & Thomas Kodadek1
- Departments of Internal Medicine and Molecular Biology, University of Texas Southwestern Medical Center, 5323 Harry Hines Blvd, Dallas, Texas 75390-9185, USA
- †Present address: Center for Innovations in Medicine, Biodesign Institute, Arizona State University, 1001 S. McAllister Ave, Tempe, Arizona 85287-5001, USA
Correspondence to: Thomas Kodadek1 Correspondence and requests for materials should be addressed to T.K. (Email: Thomas.Kodadek@utsouthwestern.edu).
Abstract
Transactivator–promoter complexes are essential intermediates in the activation of eukaryotic gene expression. Recent studies of these complexes have shown that some are quite dynamic in living cells1 owing to rapid and reversible disruption of activator–promoter complexes by molecular chaperones2, 3, 4, 5, 6, or a slower, ubiquitin–proteasome-pathway-mediated turnover of DNA-bound activator7, 8, 9. These mechanisms may act to ensure continued responsiveness of activators to signalling cascades by limiting the lifetime of the active protein–DNA complex. Furthermore, the potency of some activators is compromised by proteasome inhibition, leading to the suggestion that periodic clearance of activators from a promoter is essential for high-level expression8, 10, 11, 12. Here we describe a variant of the chromatin immunoprecipitation assay that has allowed direct observation of the kinetic stability of native Gal4–promoter complexes in yeast. Under non-inducing conditions, the complex is dynamic, but on induction the Gal4–promoter complexes 'lock in' and exhibit long half-lives. Inhibition of proteasome-mediated proteolysis had little or no effect on Gal4-mediated gene expression. These studies, combined with earlier data, show that the lifetimes of different transactivator–promoter complexes in vivo can vary widely and that proteasome-mediated turnover is not a general requirement for transactivator function.
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