Nalley et al. reply

Kip Nalley, Stephen Albert Johnston and Thomas Kodadek
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Kip Nalley: UT Southwestern Medical Center, 5323 Harry Hines Boulevard, Dallas, Texas 75390-9185, USA. kodadek@scripps.edu
Stephen Albert Johnston: UT Southwestern Medical Center, 5323 Harry Hines Boulevard, Dallas, Texas 75390-9185, USA. kodadek@scripps.edu
Thomas Kodadek: UT Southwestern Medical Center, 5323 Harry Hines Boulevard, Dallas, Texas 75390-9185, USA. kodadek@scripps.edu

Nature, 2009, vol. 461, issue 7265, E8-E8

Abstract: Abstract Replying to: G. A. Collins, J. R. Lipford, R. J. Deshaies & W. P. Tansey Nature 461, 10.1038/nature08406 (2009) Proteasome-mediated turnover of some1,2, but clearly not all3,4, transcriptional activators is important for their activity. To facilitate the analysis of activator–promoter complex lifetime in vivo, a parameter relevant to this issue, we developed a competition chromatin immunoprecipitation (ChIP) assay in which binding of a native transactivator to its cognate promoters is challenged by a ligand-activated competitor protein with the same DNA-binding specificity. We applied this technique to the yeast Gal4 system5 and concluded that under non-inducing conditions (raffinose media) Gal4–promoter complexes exchange rapidly, but under inducing conditions (galactose media) the activator–promoter complexes are long-lived. Collins et al.6 report that, surprisingly, the addition of oestradiol to yeast lacking Myc–G4–ER–VP16 increased the amount of DNA co-immunoprecipitated with native Gal4.

Date: 2009
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DOI: 10.1038/nature08407

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