Fenrick R, Amann J M, Lutterbach B, Wang L, Westendorf J J, Downing J R, Hiebert S W
Department of Biochemistry, Vanderbilt University School of Medicine, Nashville, Tennessee 37232, USA.
Mol Cell Biol. 1999 Oct;19(10):6566-74. doi: 10.1128/MCB.19.10.6566.
t(12;21) is the most frequent translocation found in pediatric B-cell acute lymphoblastic leukemias. This translocation fuses a putative repressor domain from the TEL DNA-binding protein to nearly all of the AML-1B transcription factor. Here, we demonstrate that fusion of the TEL pointed domain to the GAL4 DNA-binding domain resulted in sequence-specific transcriptional repression, indicating that the pointed domain is a portable repression motif. The TEL pointed domain functioned equally well when the GAL4 DNA-binding sites were moved 600 bp from the promoter, suggesting an active mechanism of repression. This lead us to demonstrate that wild-type TEL and the t(12;21) fusion protein bind the mSin3A corepressor. In the fusion protein, both TEL and AML-1B contribute mSin3 interaction domains. Deletion mutagenesis indicated that both the TEL and AML-1B mSin3-binding domains contribute to repression by the fusion protein. While both TEL and AML-1B associate with mSin3A, TEL/AML-1B appears to bind this corepressor much more stably than either wild-type protein, suggesting a mode of action for the t(12;21) fusion protein.
t(12;21)是小儿B细胞急性淋巴细胞白血病中最常见的易位。这种易位将TEL DNA结合蛋白的一个假定的阻遏结构域与几乎所有的AML-1B转录因子融合。在这里,我们证明将TEL尖结构域与GAL4 DNA结合结构域融合会导致序列特异性转录抑制,表明尖结构域是一个可移植的抑制基序。当GAL4 DNA结合位点从启动子移至600 bp处时,TEL尖结构域同样发挥作用,提示存在一种活跃的抑制机制。这使我们证明野生型TEL和t(12;21)融合蛋白与mSin3A共抑制因子结合。在融合蛋白中,TEL和AML-1B均提供mSin3相互作用结构域。缺失诱变表明TEL和AML-1B的mSin3结合结构域均对融合蛋白的抑制作用有贡献。虽然TEL和AML-1B均与mSin3A相关联,但TEL/AML-1B似乎比任何一种野生型蛋白都更稳定地结合这种共抑制因子,提示了t(12;21)融合蛋白的一种作用模式。