Graduate School of Frontier Biosciences, Osaka University

Japanese

Regulation of RNA polymerase II activation by histone acetylation in single living cells

Journal Nature 516, 272-275 (2014)
Authors Stasevich TJ (1, 2, 3), Hayashi-Takanaka Y (1, 4, 5), Sato Y (1, 5), Maehara K (6), Ohkawa Y (4, 6), Sakata-Sogawa K (7, 8), Tokunaga M (7, 8), Nagase T (9), Nozaki N (10), McNally JG (11, 12), Kimura H (1, 4, 5)

  1. Graduate School of Frontier Biosciences, Osaka University, Osaka, 565-0871, Japan
  2. Department of Biochemistry and Molecular Biology, Colorado State University, Fort Collins, Colorado 80523-1870, USA
  3. Transcription Imaging Consortium, Janelia Farm Research Campus, Howard Hughes Medical Institute, Ashburn, Virginia 20147, USA
  4. Japan Science and Technology Agency (JST), Core Research for Evolutional Science and Technology (CREST), Kawaguchi, Saitama, 332-0012, Japan
  5. Department of Biological Sciences, Graduate School of Bioscience and Biotechnology, Tokyo Institute of Technology, Yokohama, 226-8501, Japan
  6. Department of Advanced Medical Initiatives, Faculty of Medicine, Kyushu University, Fukuoka, 812-8582, Japan
  7. Department of Biological Information, Graduate School of Bioscience and Biotechnology, Tokyo Institute of Technology, Yokohama, 226-8501, Japan
  8. RIKEN Center for Integrative Medical Sciences (IMS), Yokohama, 230-0045, Japan
  9. Department of Biotechnology Research, Kazusa DNA Research Institute, Chiba, 292-0818, Japan
  10. Mab Institute Inc., Sapporo, 001-0021, Japan
  11. Laboratory of Receptor Biology and Gene Expression, National Cancer Institute, National Institutes of Health, Bethesda, Maryland 20892, USA
  12. Institute for Soft Matter and Functional Materials, Helmholtz Zentrum Berlin, Berlin, 14109, Germany
Title Regulation of RNA polymerase II activation by histone acetylation in single living cells
PubMed 25252976
Laboratory Nuclear Dynamics Group 〈Prof. Hiraoka〉
Description In eukaryotic cells, post-translational histone modifications have an important role in gene regulation. Starting with early work on histone acetylation, a variety of residue-specific modifications have now been linked to RNA polymerase II (RNAP2) activity, but it remains unclear if these markers are active regulators of transcription or just passive byproducts. This is because studies have traditionally relied on fixed cell populations, meaning temporal resolution is limited to minutes at best, and correlated factors may not actually be present in the same cell at the same time. Complementary approaches are therefore needed to probe the dynamic interplay of histone modifications and RNAP2 with higher temporal resolution in single living cells. Here we address this problem by developing a system to track residue-specific histone modifications and RNAP2 phosphorylation in living cells by fluorescence microscopy. This increases temporal resolution to the tens-of-seconds range. Our single-cell analysis reveals histone H3 lysine-27 acetylation at a gene locus can alter downstream transcription kinetics by as much as 50%, affecting two temporally separate events. First acetylation enhances the search kinetics of transcriptional activators, and later the acetylation accelerates the transition of RNAP2 from initiation to elongation. Signatures of the latter can be found genome-wide using chromatin immunoprecipitation followed by sequencing. We argue that this regulation leads to a robust and potentially tunable transcriptional response.