Biophysical characterization of histone H3.3 K27M point mutation

Szabolcs Hetey, Beáta Boros-Oláh, Tímea Kuik-Rózsa, Qiuzhen Li, Z. Karányi, Z. Szabó, Jason Roszik, Nikoletta Szalóki, G. Vámosi, Katalin Tóth, Lóránt Székvölgyi

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Abstract

Lysine 27 to methionine (K27 M) mutation of the histone variant H3.3 drives the formation of an aggressive glioblastoma multiforme tumor in infants. Here we analyzed how the methionine substitution alters the stability of H3.3 nucleosomes in vitro and modifies its kinetic properties in live cells. We also determined whether the presence of mutant nucleosomes perturbed the mobility of the PRC2 subunit Ezh2 (enhancer-of-zeste homolog 2). We found that K27 M nucleosomes maintained the wild-type molecular architecture both at the level of bulk histones and single nucleosomes and followed similar diffusion kinetics to wild-type histones in live cells. Nevertheless, we observed a remarkable differential recovery of Ezh2 in response to transcriptional stress that was accompanied by a faster diffusion rate of the mobile fraction of Ezh2 and a significantly increased immobile fraction, suggesting tighter chromatin binding of Ezh2 upon transcription inhibition. The differential recovery of Ezh2 was dependent on transcription, however, it was independent from K27 M mutation status. These biophysical characteristics shed more light on the mechanism of histone H3.3 K27M in glioma genesis in relation to the kinetic properties of Ezh2.

Original languageEnglish
Pages (from-to)868-875
Number of pages8
JournalBiochemical and Biophysical Research Communications
Volume490
Issue number3
DOIs
Publication statusPublished - Aug 26 2017

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Keywords

  • Ezh2
  • Glioblastoma
  • H3.3 K27M

ASJC Scopus subject areas

  • Biophysics
  • Biochemistry
  • Molecular Biology
  • Cell Biology

Cite this

Hetey, S., Boros-Oláh, B., Kuik-Rózsa, T., Li, Q., Karányi, Z., Szabó, Z., Roszik, J., Szalóki, N., Vámosi, G., Tóth, K., & Székvölgyi, L. (2017). Biophysical characterization of histone H3.3 K27M point mutation. Biochemical and Biophysical Research Communications, 490(3), 868-875. https://doi.org/10.1016/j.bbrc.2017.06.133