3D orbital tracking; RNA synthesis; bursting kinetics; circadian cycle; glucocorticoid receptor; high-throughput imaging; single cell; single molecule tracking; transcription dynamics; ultradian hormone stimulation; Glucocorticoids; Receptors, Glucocorticoid; RNA; Animals; Glucocorticoids/pharmacology; Mice; RNA/biosynthesis; RNA/genetics; Receptors, Glucocorticoid/metabolism; Transcription, Genetic/drug effects; Promoter Regions, Genetic; Transcription Initiation Site
Abstract :
[en] Genes are transcribed in a discontinuous pattern referred to as RNA bursting, but the mechanisms regulating this process are unclear. Although many physiological signals, including glucocorticoid hormones, are pulsatile, the effects of transient stimulation on bursting are unknown. Here we characterize RNA synthesis from single-copy glucocorticoid receptor (GR)-regulated transcription sites (TSs) under pulsed (ultradian) and constant hormone stimulation. In contrast to constant stimulation, pulsed stimulation induces restricted bursting centered around the hormonal pulse. Moreover, we demonstrate that transcription factor (TF) nuclear mobility determines burst duration, whereas its bound fraction determines burst frequency. Using 3D tracking of TSs, we directly correlate TF binding and RNA synthesis at a specific promoter. Finally, we uncover a striking co-bursting pattern between TSs located at proximal and distal positions in the nucleus. Together, our data reveal a dynamic interplay between TF mobility and RNA bursting that is responsive to stimuli strength, type, modality, and duration.
Disciplines :
Biochemistry, biophysics & molecular biology
Author, co-author :
Stavreva, Diana A; Laboratory of Receptor Biology and Gene Expression, 41 Library Drive, Center for Cancer Research, NCI, NIH, Bethesda, MD 20892-5055, USA. Electronic address: stavrevd@mail.nih.gov
Garcia, David A; Laboratory of Receptor Biology and Gene Expression, 41 Library Drive, Center for Cancer Research, NCI, NIH, Bethesda, MD 20892-5055, USA, Department of Physics and Institute for Physical Science and Technology, University of Maryland, College Park, MD 20742, USA
Fettweis, Grégory ; Université de Liège - ULiège > Département des sciences de la vie > Génétique et biologie moléculaires animales ; Laboratory of Receptor Biology and Gene Expression, 41 Library Drive, Center for Cancer Research, NCI, NIH, Bethesda, MD 20892-5055, USA
Gudla, Prabhakar R; Laboratory of Receptor Biology and Gene Expression, 41 Library Drive, Center for Cancer Research, NCI, NIH, Bethesda, MD 20892-5055, USA
Zaki, George F; High Performance Computing Group, Frederick National Laboratory for Cancer Research, Frederick, MD, USA
Soni, Vikas; Laboratory of Receptor Biology and Gene Expression, 41 Library Drive, Center for Cancer Research, NCI, NIH, Bethesda, MD 20892-5055, USA
McGowan, Andrew; Laboratory of Receptor Biology and Gene Expression, 41 Library Drive, Center for Cancer Research, NCI, NIH, Bethesda, MD 20892-5055, USA
Williams, Geneva; Laboratory of Receptor Biology and Gene Expression, 41 Library Drive, Center for Cancer Research, NCI, NIH, Bethesda, MD 20892-5055, USA
Huynh, Anh; Department of Physics and Graduate Program in Biomolecular Science, Boise State University, Boise, ID 83725, USA
Palangat, Murali; Laboratory of Receptor Biology and Gene Expression, 41 Library Drive, Center for Cancer Research, NCI, NIH, Bethesda, MD 20892-5055, USA
Schiltz, R Louis; Laboratory of Receptor Biology and Gene Expression, 41 Library Drive, Center for Cancer Research, NCI, NIH, Bethesda, MD 20892-5055, USA
Johnson, Thomas A; Laboratory of Receptor Biology and Gene Expression, 41 Library Drive, Center for Cancer Research, NCI, NIH, Bethesda, MD 20892-5055, USA
Presman, Diego M; Laboratory of Receptor Biology and Gene Expression, 41 Library Drive, Center for Cancer Research, NCI, NIH, Bethesda, MD 20892-5055, USA
Ferguson, Matthew L; Department of Physics and Graduate Program in Biomolecular Science, Boise State University, Boise, ID 83725, USA
Pegoraro, Gianluca; Laboratory of Receptor Biology and Gene Expression, 41 Library Drive, Center for Cancer Research, NCI, NIH, Bethesda, MD 20892-5055, USA
Upadhyaya, Arpita; Department of Physics and Institute for Physical Science and Technology, University of Maryland, College Park, MD 20742, USA
Hager, Gordon L; Laboratory of Receptor Biology and Gene Expression, 41 Library Drive, Center for Cancer Research, NCI, NIH, Bethesda, MD 20892-5055, USA. Electronic address: hagerg@exchange.nih.gov
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