Abstract
With the objective of returning cells to their undifferentiated state through alteration of epigenetic states, small molecules have been used that specifically inhibit proteins involved in sustaining the epigenetic system. However, this chemical-based approach can cause chaotic epigenomic states due to random actions of the inhibitors. We investigated whether JHDM3A/JMJD2A, a trimethylated histone H3-lysine 9 (H3K9me3)-specific demethylase, could function as an effector molecule to selectively demethylate target chromatin, with the aid of a guide protein to serve as a delivery vehicle. JHDM3A, which normally locates in euchromatin, spread out to heterochromatin when it was fused to heterochromatin protein-1α (HP1α) or HP1β; in these cells, demethylation efficiency was also markedly increased. Two truncated modules, JHDM3AGFP 406 and JHDM3AGFP 701, had contrasting modes and efficiencies of H3K9me3 demethylation; JHDM3A GFP 406 showed a very uniform rate (∼80%) of demethylation, whereas JHDM3AGFP 701 had a broad methylation range of 4-80%. The methylation values were highly dependent on the presence of the guide proteins OCT4, CTCF, and HP1. Chromatin immunoprecipitation detected reduced H3K9me3 levels at OCT4 regulatory loci in the cells expressing OCT4-tagged JHDM3AGFP 701. Derepression of the Sox2 gene was observed in JHDM3AGFP 701 OCT4-expressing cells, but not in cells that expressed the JHDM3A GFP 701 module alone. JHDM3AGFP 701-assisted OCT4 more efficiently turned on stem cell-related microRNAs than GFP-OCT4 itself. These results suggest that JHDM3A GFP 701 is a suitable catalytic module that can be targeted, under the control of a guide protein, to specific loci where the chromatin H3K9me3 status and the milieu of gene expression are to be modified.
| Original language | English |
|---|---|
| Pages (from-to) | 4461-4470 |
| Number of pages | 10 |
| Journal | Journal of Biological Chemistry |
| Volume | 286 |
| Issue number | 6 |
| DOIs | |
| Publication status | Published - 2011 Feb 11 |
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ASJC Scopus subject areas
- Biochemistry
- Cell Biology
- Molecular Biology
- Medicine(all)
Cite this
JHDM3A module as an effector molecule in guide-directed modification of target chromatin. / Jeong, Young Sun; Park, Jung Sun; Ko, Yong; Kang, Yong Kook.
In: Journal of Biological Chemistry, Vol. 286, No. 6, 11.02.2011, p. 4461-4470.Research output: Contribution to journal › Article
}
TY - JOUR
T1 - JHDM3A module as an effector molecule in guide-directed modification of target chromatin
AU - Jeong, Young Sun
AU - Park, Jung Sun
AU - Ko, Yong
AU - Kang, Yong Kook
PY - 2011/2/11
Y1 - 2011/2/11
N2 - With the objective of returning cells to their undifferentiated state through alteration of epigenetic states, small molecules have been used that specifically inhibit proteins involved in sustaining the epigenetic system. However, this chemical-based approach can cause chaotic epigenomic states due to random actions of the inhibitors. We investigated whether JHDM3A/JMJD2A, a trimethylated histone H3-lysine 9 (H3K9me3)-specific demethylase, could function as an effector molecule to selectively demethylate target chromatin, with the aid of a guide protein to serve as a delivery vehicle. JHDM3A, which normally locates in euchromatin, spread out to heterochromatin when it was fused to heterochromatin protein-1α (HP1α) or HP1β; in these cells, demethylation efficiency was also markedly increased. Two truncated modules, JHDM3AGFP 406 and JHDM3AGFP 701, had contrasting modes and efficiencies of H3K9me3 demethylation; JHDM3A GFP 406 showed a very uniform rate (∼80%) of demethylation, whereas JHDM3AGFP 701 had a broad methylation range of 4-80%. The methylation values were highly dependent on the presence of the guide proteins OCT4, CTCF, and HP1. Chromatin immunoprecipitation detected reduced H3K9me3 levels at OCT4 regulatory loci in the cells expressing OCT4-tagged JHDM3AGFP 701. Derepression of the Sox2 gene was observed in JHDM3AGFP 701 OCT4-expressing cells, but not in cells that expressed the JHDM3A GFP 701 module alone. JHDM3AGFP 701-assisted OCT4 more efficiently turned on stem cell-related microRNAs than GFP-OCT4 itself. These results suggest that JHDM3A GFP 701 is a suitable catalytic module that can be targeted, under the control of a guide protein, to specific loci where the chromatin H3K9me3 status and the milieu of gene expression are to be modified.
AB - With the objective of returning cells to their undifferentiated state through alteration of epigenetic states, small molecules have been used that specifically inhibit proteins involved in sustaining the epigenetic system. However, this chemical-based approach can cause chaotic epigenomic states due to random actions of the inhibitors. We investigated whether JHDM3A/JMJD2A, a trimethylated histone H3-lysine 9 (H3K9me3)-specific demethylase, could function as an effector molecule to selectively demethylate target chromatin, with the aid of a guide protein to serve as a delivery vehicle. JHDM3A, which normally locates in euchromatin, spread out to heterochromatin when it was fused to heterochromatin protein-1α (HP1α) or HP1β; in these cells, demethylation efficiency was also markedly increased. Two truncated modules, JHDM3AGFP 406 and JHDM3AGFP 701, had contrasting modes and efficiencies of H3K9me3 demethylation; JHDM3A GFP 406 showed a very uniform rate (∼80%) of demethylation, whereas JHDM3AGFP 701 had a broad methylation range of 4-80%. The methylation values were highly dependent on the presence of the guide proteins OCT4, CTCF, and HP1. Chromatin immunoprecipitation detected reduced H3K9me3 levels at OCT4 regulatory loci in the cells expressing OCT4-tagged JHDM3AGFP 701. Derepression of the Sox2 gene was observed in JHDM3AGFP 701 OCT4-expressing cells, but not in cells that expressed the JHDM3A GFP 701 module alone. JHDM3AGFP 701-assisted OCT4 more efficiently turned on stem cell-related microRNAs than GFP-OCT4 itself. These results suggest that JHDM3A GFP 701 is a suitable catalytic module that can be targeted, under the control of a guide protein, to specific loci where the chromatin H3K9me3 status and the milieu of gene expression are to be modified.
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U2 - 10.1074/jbc.M110.176040
DO - 10.1074/jbc.M110.176040
M3 - Article
VL - 286
SP - 4461
EP - 4470
JO - Journal of Biological Chemistry
JF - Journal of Biological Chemistry
SN - 0021-9258
IS - 6
ER -