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首页> 外文期刊>Journal of cellular biochemistry. >Identification of novel protein/DNA interactions within the promoter of the bone-related transcription factor Runx2/Cbfa1.
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Identification of novel protein/DNA interactions within the promoter of the bone-related transcription factor Runx2/Cbfa1.

机译：鉴定骨相关转录因子Runx2 / Cbfa1启动子中的新型蛋白质/ DNA相互作用。

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摘要

The runt homology transcription factor Runx2/Cbfa1 is essential for bone development and osteoblast differentiation. Regulatory mechanisms that govern Runx2 transcription in osteoblasts define the osteogenic pathways that control skeletal development. In this study, we systematically examined transcription factor binding within the upstream Runx2 P1 promoter, which regulates expression of the bone-related Runx2 factor. We identified two novel protein/DNA interactions that are mediated by sequence specific factors, based on cross-competition experiments, point mutations, and gel-shift immunoassays. One complex recognizes a non-canonical Runx2 site, whereas the other factor binds to a palindromic sequence. Site-directed mutagenesis of the novel Runx2 motif (5'TCCCAC3') within the 0.6 kb rat Runx2 promoter reduces transcription by 2-fold, indicating that this site supports enhancement of Runx2 promoter activity. Mutation of the palindromic motif (5'AGTACT3') results in a 2-3-fold activation of the Runx2 promoter, demonstrating that the wild type sequence contributes to transcriptional repression. These studies, together with our previous findings of auto-suppression of the Runx2 promoter and negative regulation by 1,25(OH)(2) Vitamin D3, suggest that physiological control of Runx2 gene expression is mediated by a series of intricate regulatory mechanisms.
机译：矮子同源转录因子Runx2 / Cbfa1对骨骼发育和成骨细胞分化至关重要。在成骨细胞中控制Runx2转录的调控机制定义了控制骨骼发育的成骨途径。在这项研究中，我们系统地检查了上游Runx2 P1启动子内的转录因子结合，该启动子调节与骨相关的Runx2因子的表达。我们确定了两种新的蛋白质/ DNA相互作用，这些相互作用是由序列特异性因子介导的，基于交叉竞争实验，点突变和凝胶移位免疫测定。一个复合体识别一个非规范的Runx2位点，而另一个复合体则结合回文序列。在0.6 kb大鼠Runx2启动子内对新型Runx2基序（5'TCCCAC3'）进行定点诱变可将转录降低2倍，表明该位点支持Runx2启动子活性的增强。回文基序（5'AGTACT3'）的突变导致Runx2启动子的2到3倍激活，表明野生型序列有助于转录抑制。这些研究，加上我们先前对Runx2启动子的自动抑制和1,25（OH）（2）维生素D3负调控的发现，表明Runx2基因表达的生理控制是由一系列复杂的调控机制介导的。

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