Identification of an evolutionarily conserved domain in human lens epithelium-derived growth factor/transcriptional co-activator p75 (LEDGF/p75) that binds HIV-1 integrase
Identification of an evolutionarily conserved domain in human lens epithelium-derived growth factor/transcriptional co-activator p75 (LEDGF/p75) that binds HIV-1 integrase
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DOI：
10.1074/jbc.m406307200
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发表时间：
2004-11-19
影响因子：
4.8
通讯作者：
Engelman, A
中科院分区：
文献类型：
--
作者：
Cherepanov, P;Devroe, E;Engelman, A
人类晶状体上皮源性生长因子/转录共激活因子p75(LEDGF/p75)蛋白是新近发现的人类细胞中HIV-1整合酶(IN)的结合伙伴。在这项工作中，我们使用生化和生物信息学方法来定义LEDGF/p75的结构域组织。利用有限的蛋白分解和缺失突变，我们发现该蛋白含有一对进化保守的结构域，约占ITS序列的35%。鉴于N-末端PWWP结构域以前已被识别，第二个结构域是新的。它由类似于80个氨基酸残基组成，是与HIV-1 IN结合的必要条件和充分条件。值得注意的是，整合酶结合结构域(IBD)并不是LEDGF/p75所特有的，作为第二种人类蛋白质，肝癌衍生生长因子相关蛋白2(HRP2)包含一个同源序列。在体外GST下拉实验中，LEDGF/p75和HRP2 IBD与HIV-1IN强烈结合，每个全长蛋白在体外都能有效地刺激HIV-1IN的活性。LEDGF/p75和HRP2被预测共享相似的域组织，并具有明显的进化和可能的功能关系。
Human lens epithelium-derived growth factor/transcriptional co-activator p75 (LEDGF/p75) protein was recently identified as a binding partner for HIV-1 integrase (IN) in human cells. In this work, we used biochemical and bioinformatic approaches to define the domain organization of LEDGF/p75. Using limited proteolysis and deletion mutagenesis we show that the protein contains a pair of evolutionarily conserved domains, assuming about 35% of its sequence. Whereas the N-terminal PWWP domain had been recognized previously, the second domain is novel. It is comprised of similar to80 amino acid residues and is both necessary and sufficient for binding to HIV-1 IN. Strikingly, the integrase binding domain (IBD) is not unique to LEDGF/p75, as a second human protein, hepatoma-derived growth factor-related protein 2 (HRP2), contains a homologous sequence. LEDGF/p75 and HRP2 IBDs avidly bound HIV-1 IN in an in vitro GST pull-down assay and each full-length protein potently stimulated HIV-1 IN activity in vitro. LEDGF/p75 and HRP2 are predicted to share a similar domain organization and have an evident evolutionary and likely functional relationship.