RRC ID 48774
Author Kurita K, Sakamoto T, Yagi N, Sakamoto Y, Ito A, Nishino N, Sako K, Yoshida M, Kimura H, Seki M, Matsunaga S.
Title Live imaging of H3K9 acetylation in plant cells.
Journal Sci Rep
Abstract Proper regulation of histone acetylation is important in development and cellular responses to environmental stimuli. However, the dynamics of histone acetylation at the single-cell level remains poorly understood. Here we established a transgenic plant cell line to track histone H3 lysine 9 acetylation (H3K9ac) with a modification-specific intracellular antibody (mintbody). The H3K9ac-specific mintbody fused to the enhanced green fluorescent protein (H3K9ac-mintbody-GFP) was introduced into tobacco BY-2 cells. We successfully demonstrated that H3K9ac-mintbody-GFP interacted with H3K9ac in vivo. The ratio of nuclear/cytoplasmic H3K9ac-mintbody-GFP detected in quantitative analysis reflected the endogenous H3K9ac levels. Under chemically induced hyperacetylation conditions with histone deacetylase inhibitors including trichostatin A, Ky-2 and Ky-14, significant enhancement of H3K9ac was detected by H3K9ac-mintbody-GFP dependent on the strength of inhibitors. Conversely, treatment with a histone acetyltransferase inhibitor, C646 caused a reduction in the nuclear to cytoplasmic ratio of H3K9ac-mintbody-GFP. Using this system, we assessed the environmental responses of H3K9ac and found that cold and salt stresses enhanced H3K9ac in tobacco BY-2 cells. In addition, a combination of H3K9ac-mintbody-GFP with 5-ethynyl-2'-deoxyuridine labelling confirmed that H3K9ac level is constant during interphase.
Volume 7
Pages 45894
Published 2017-4-18
DOI 10.1038/srep45894
PII srep45894
PMID 28418019
PMC PMC5394682
MeSH Acetylation / drug effects Antibodies / immunology Antibodies / metabolism Benzoates / pharmacology Green Fluorescent Proteins / chemistry Histone Acetyltransferases / genetics* Histone Acetyltransferases / immunology Histone Acetyltransferases / ultrastructure Histone Deacetylase Inhibitors / pharmacology Histones / immunology Plant Cells / metabolism Plant Cells / ultrastructure* Plants, Genetically Modified Protein Processing, Post-Translational / genetics* Pyrazoles / pharmacology Tobacco / genetics Tobacco / growth & development Tobacco / metabolism*
IF 3.998
Times Cited 3
WOS Category MULTIDISCIPLINARY SCIENCES
Resource
Arabidopsis / Cultured plant cells, genes rpc00001