Spensberger Dominik, Vermeulen Michiel, Le Guezennec Xavier, Beekman Renee, van Hoven Antoinette, Bindels Eric, Stunnenberg Henk, Delwel Ruud
Department of Hematology, Erasmus University Medical Center, Rotterdam 3015 GE, The Netherlands.
Biochemistry. 2008 Jun 17;47(24):6418-26. doi: 10.1021/bi800267f. Epub 2008 May 24.
The ecotropic viral integration site 1 ( Evi1) gene encodes a putative transcription regulator, which is aberrantly expressed in acute myeloid leukemias (AML) with chromosomal abnormalities involving the 3q26 locus. Repression and activation of transcriptional control have been reported, but it is currently unclear how Evi1 may evoke these opposing effects. Using a yeast two-hybrid screen, we identified a novel binding partner of Evi1, i.e., methyl binding domain 3b (Mbd3b) protein, a member of the Mi-2/NuRD histone deacetylase complex. Applying in vitro and in vivo assays, we found that Evi1 interacts with Mbd3b but not with other MBD family members Mbd1, -2, and -4 or MeCP2. We show that interaction of Evi1 with Mbd3 requires 40 amino acids that are adjacent and downstream of the methyl binding domain (MBD). We further demonstrate that the first three zinc fingers of Evi1 are needed for Mbd3 interaction. Evi1 acts as a transcriptional repressor when recruited to an active promoter, yet when present in the Mi-2/NuRD complex through Mbd3 interaction, it inhibits the histone deacetylation function of this multiprotein structure. Our data may in part explain how Evi1 could act as a repressor as well as an activator of transcription.
嗜亲性病毒整合位点1(Evi1)基因编码一种假定的转录调节因子,该因子在伴有涉及3q26位点染色体异常的急性髓性白血病(AML)中异常表达。虽然已有转录抑制和激活的报道,但目前尚不清楚Evi1如何引发这些相反的效应。通过酵母双杂交筛选,我们鉴定出Evi1的一个新的结合伴侣,即甲基结合结构域3b(Mbd3b)蛋白,它是Mi-2/NuRD组蛋白去乙酰化酶复合物的成员。通过体外和体内试验,我们发现Evi1与Mbd3b相互作用,但不与其他MBD家族成员Mbd1、-2和-4或MeCP2相互作用。我们表明,Evi1与Mbd3的相互作用需要甲基结合结构域(MBD)相邻及下游的40个氨基酸。我们进一步证明,Evi1的前三个锌指对于与Mbd3的相互作用是必需的。当Evi1被招募到一个活性启动子时,它作为转录抑制因子起作用,然而当通过与Mbd3相互作用存在于Mi-2/NuRD复合物中时,它会抑制这种多蛋白结构的组蛋白去乙酰化功能。我们的数据可能部分解释了Evi1如何既作为转录抑制因子又作为转录激活因子发挥作用。