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

Timothy J. Stasevich, Yoko Hayashi-Takanaka, Yuko Sato, Kazumitsu Maehara, Yasuyuki Ohkawa, Kumiko Sakata-Sogawa, Makio Tokunaga, Takahiro Nagase, Naohito Nozaki, James G. McNally, Hiroshi Kimura

Research output: Contribution to journalArticle

120 Citations (Scopus)

Abstract

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.

Original languageEnglish
Pages (from-to)272-275
Number of pages4
JournalNature
Volume516
Issue number7530
DOIs
Publication statusPublished - Dec 11 2014

All Science Journal Classification (ASJC) codes

  • General

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    Stasevich, T. J., Hayashi-Takanaka, Y., Sato, Y., Maehara, K., Ohkawa, Y., Sakata-Sogawa, K., Tokunaga, M., Nagase, T., Nozaki, N., McNally, J. G., & Kimura, H. (2014). Regulation of RNA polymerase II activation by histone acetylation in single living cells. Nature, 516(7530), 272-275. https://doi.org/10.1038/nature13714