Single-molecule analysis of steroid receptor and cofactor action in living cells

Ville Paakinaho, Diego M. Presman, David A. Ball, Thomas A. Johnson, R. Louis Schiltz, Peter Levitt, Davide Mazza, Tatsuya Morisaki, Tatiana S. Karpova and Gordon L. Hager ()
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Ville Paakinaho: Laboratory of Receptor Biology and Gene Expression, National Cancer Institute, National Institutes of Health
Diego M. Presman: Laboratory of Receptor Biology and Gene Expression, National Cancer Institute, National Institutes of Health
David A. Ball: Laboratory of Receptor Biology and Gene Expression, National Cancer Institute, National Institutes of Health
Thomas A. Johnson: Laboratory of Receptor Biology and Gene Expression, National Cancer Institute, National Institutes of Health
R. Louis Schiltz: Laboratory of Receptor Biology and Gene Expression, National Cancer Institute, National Institutes of Health
Peter Levitt: Laboratory of Receptor Biology and Gene Expression, National Cancer Institute, National Institutes of Health
Davide Mazza: Istituto Scientifico Ospedale San Raffaele, Centro di Imaging Sperimentale e Università Vita-Salute San Raffaele
Tatsuya Morisaki: Laboratory of Receptor Biology and Gene Expression, National Cancer Institute, National Institutes of Health
Tatiana S. Karpova: Laboratory of Receptor Biology and Gene Expression, National Cancer Institute, National Institutes of Health
Gordon L. Hager: Laboratory of Receptor Biology and Gene Expression, National Cancer Institute, National Institutes of Health

Nature Communications, 2017, vol. 8, issue 1, 1-14

Abstract: Abstract Population-based assays have been employed extensively to investigate the interactions of transcription factors (TFs) with chromatin and are often interpreted in terms of static and sequential binding. However, fluorescence microscopy techniques reveal a more dynamic binding behaviour of TFs in live cells. Here we analyse the strengths and limitations of in vivo single-molecule tracking and performed a comprehensive analysis on the intranuclear dwell times of four steroid receptors and a number of known cofactors. While the absolute residence times estimates can depend on imaging acquisition parameters due to sampling bias, our results indicate that only a small proportion of factors are specifically bound to chromatin at any given time. Interestingly, the glucocorticoid receptor and its cofactors affect each other’s dwell times in an asymmetric manner. Overall, our data indicate transient rather than stable TF-cofactors chromatin interactions at response elements at the single-molecule level.

Date: 2017
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DOI: 10.1038/ncomms15896

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