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首页> 外文期刊>Molecular and Cellular Biology >The Capacity of Polyomavirus Enhancer Binding Protein 2αB (AML1/Cbfa2) To Stimulate Polyomavirus DNA Replication Is Related to Its Affinity for the Nuclear Matrix
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The Capacity of Polyomavirus Enhancer Binding Protein 2αB (AML1/Cbfa2) To Stimulate Polyomavirus DNA Replication Is Related to Its Affinity for the Nuclear Matrix

机译：多瘤病毒增强子结合蛋白2αB（AML1 / Cbfa2）刺激多瘤病毒DNA复制的能力与其对核基质的亲和力有关

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摘要

The nuclear matrix is thought to play an important role in the DNA replication of eukaryotic cells, although direct evidence for such a role is still lacking. A nuclear matrix-associated transcription factor, polyomavirus (Py) enhancer binding protein 2αB1 (PEBP2αB1) (AML1/Cbfa2), was found to stimulate Py replication through its cognate binding site. The minimal replication activation domain (RAD) was identified between amino acid (aa) 302 and aa 371 by using a fusion protein containing the GAL4 DNA binding domain (GAL4-RAD). In addition, the region showed affinity for the nuclear matrix and, on the basis of competition studies, binding activity for one or more proteins involved in the initiation of Py DNA replication. A leukemogenic chimeric protein, AML1/ETO(MTG8), which does not contain this region of PEBP2αB1/AML1, was also localized in the nuclear matrix fraction and competed for nuclear matrix association with PEBP2αB1 and GAL4-RAD. Moreover, AML1/ETO inhibited Py DNA replication stimulated by PEBP2αB1 and GAL4-RAD. The inhibition was specific for replication mediated by PEBP2αB1 and GAL4-RAD, and proportional to the degree of loss of these activators from the nuclear matrix, suggesting a requirement for nuclear matrix targeting in the stimulation of Py DNA replication by RAD. These results are the first to suggest a molecular link between the initiation of DNA replication and the nuclear matrix compartment.
机译：尽管仍缺乏直接证据证明核基质在真核细胞的DNA复制中起着重要作用。发现与核基质相关的转录因子多瘤病毒（Py）增强子结合蛋白2αB1（PEBP2αB1）（AML1 / Cbfa2）可以通过其同源结合位点刺激Py复制。通过使用含有GAL4 DNA结合域（GAL4-RAD）的融合蛋白，在氨基酸（aa）302和氨基酸371之间鉴定了最小复制激活域（RAD）。此外，该区域显示出对核基质的亲和力，并且在竞争研究的基础上，显示了与一种或多种参与Py DNA复制起始的蛋白质的结合活性。不包含PEBP2αB1/ AML1此区域的致白血病嵌合蛋白AML1 / ETO（MTG8）也位于核基质组分中，并与PEBP2αB1和GAL4-RAD竞争核基质缔合。此外，AML1 / ETO抑制了PEBP2αB1和GAL4-RAD刺激的Py DNA复制。抑制作用对PEBP2αB1和GAL4-RAD介导的复制具有特异性，并且与这些活化剂从核基质中流失的程度成比例，这表明在RAD刺激Py DNA复制中需要靶向核基质。这些结果首次表明DNA复制的起始与核基质区室之间存在分子联系。

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