RRC ID |
48774
|
著者 |
Kurita K, Sakamoto T, Yagi N, Sakamoto Y, Ito A, Nishino N, Sako K, Yoshida M, Kimura H, Seki M, Matsunaga S.
|
タイトル |
Live imaging of H3K9 acetylation in plant cells.
|
ジャーナル |
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.
|
巻・号 |
7
|
ページ |
45894
|
公開日 |
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
Nitrobenzenes
Plant Cells / metabolism
Plant Cells / ultrastructure*
Plants, Genetically Modified
Protein Processing, Post-Translational / genetics*
Pyrazoles / pharmacology
Pyrazolones
Tobacco / genetics
Tobacco / growth & development
Tobacco / metabolism*
|
IF |
3.998
|
引用数 |
3
|
WOS 分野
|
MULTIDISCIPLINARY SCIENCES
|
リソース情報 |
シロイヌナズナ / 植物培養細胞・遺伝子 |
rpc00001 |