Analysis of the subcellular localization of the human histone methyltransferase SETDB1
Analysis of the subcellular localization of the human histone methyltransferase SETDB1
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DOI：
10.1016/j.bbrc.2015.08.065
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发表时间：
2015-10-02
影响因子：
3.1
通讯作者：
Doi, Takefumi
中科院分区：
文献类型：
--
作者：
Tachibana, Keisuke;Gotoh, Eiko;Doi, Takefumi
SET结构域，分叉1（SETDB 1）是一种组蛋白甲基转移酶，甲基化组蛋白H3上的赖氨酸9。虽然了解蛋白质的定位对阐明其生理功能很重要，但对人SETDB 1的亚细胞定位知之甚少。在本研究中，为了研究hSETDB 1的亚细胞定位，我们建立了组成型表达增强型绿色荧光蛋白融合hSETDB 1的人细胞系。然后，我们产生了针对hSETDB 1蛋白的单克隆抗体。外源性和内源性hSETDB 1的表达主要在各种人细胞系的细胞质中观察到。用核输出抑制剂来普霉素B和蛋白酶体抑制剂MG 132联合处理导致hSETDB 1在细胞核中积累。这些发现表明，hSETDB 1，定位在细胞核中，可能会经历降解的蛋白酶体和出口到胞质溶胶，导致其检测主要在胞质溶胶。(C)2015 Elsevier Inc. All rights reserved.
SET domain, bifurcated 1 (SETDB1) is a histone methyltransferase that methylates lysine 9 on histone H3. Although it is important to know the localization of proteins to elucidate their physiological function, little is known of the subcellular localization of human SETDB1. In the present study, to investigate the subcellular localization of hSETDB1, we established a human cell line constitutively expressing enhanced green fluorescent protein fused to hSETDB1. We then generated a monoclonal antibody against the hSETDB1 protein. Expression of both exogenous and endogenous hSETDB1 was observed mainly in the cytoplasm of various human cell lines. Combined treatment with the nuclear export inhibitor leptomycin B and the proteasome inhibitor MG132 led to the accumulation of hSETDB1 in the nucleus. These findings suggest that hSETDB1, localized in the nucleus, might undergo degradation by the proteasome and be exported to the cytosol, resulting in its detection mainly in the cytosol. (C) 2015 Elsevier Inc. All rights reserved.