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SATB2的分离与鉴定,一种在大鼠大脑中以发育和细胞特异性方式表达的新型富含AT的DNA结合蛋白。

Isolation and characterization of SATB2, a novel AT-rich DNA binding protein expressed in development- and cell-specific manner in the rat brain.

作者信息

Szemes Marianna, Gyorgy Andrea, Paweletz Cloud, Dobi Albert, Agoston Denes V

机构信息

Department of Anatomy, Physiology and Genetics, USUHS, Bethesda, MD 20814, USA.

出版信息

Neurochem Res. 2006 Feb;31(2):237-46. doi: 10.1007/s11064-005-9012-8. Epub 2006 Apr 4.

Abstract

AT-rich DNA elements play an important role in regulating cell-specific gene expression. One of the AT-rich DNA binding proteins, SATB1 is a novel type of transcription factor that regulates gene expression in the hematopoietic lineage through chromatin modification. Using DNA-affinity purification followed by mass spectrometry we identified and isolated a related protein, SATB2 from the developing rat cerebral cortex. SATB2 shows homology to SATB1 and the rat protein is practically identical to the mouse and human SATB2. Using competitive EMSA, we show that recombinant SATB2 protein binds with high affinity and specificity to AT-rich dsDNA. Using RT-PCR, Western analysis and immunohistochemistry we demonstrate that SATB2 expression is restricted to a subset of postmitotic, differentiating neurons in the rat neocortex at ages E16 and P4. We suggest that similar to its homologue SATB1, SATB2 is also involved in regulating gene expression through altering chromatin structure in differentiating cortical neurons.

摘要

富含AT的DNA元件在调节细胞特异性基因表达中发挥着重要作用。富含AT的DNA结合蛋白之一,SATB1是一种新型转录因子,可通过染色质修饰调节造血谱系中的基因表达。我们通过DNA亲和纯化后进行质谱分析,从发育中的大鼠大脑皮层中鉴定并分离出一种相关蛋白SATB2。SATB2与SATB1具有同源性,大鼠蛋白与小鼠和人类的SATB2几乎相同。通过竞争性电泳迁移率变动分析,我们发现重组SATB2蛋白能以高亲和力和特异性结合富含AT的双链DNA。利用逆转录聚合酶链反应、蛋白质免疫印迹分析和免疫组织化学,我们证明在胚胎第16天和出生后第4天的大鼠新皮层中,SATB2的表达局限于有丝分裂后正在分化的神经元亚群。我们认为,与其同源物SATB1类似,SATB2也通过改变分化中的皮层神经元的染色质结构参与调节基因表达。

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