Kho C J, Huggins G S, Endege W O, Patterson C, Jain M K, Lee M E, Haber E
Cardiovascular Biology Laboratory, Harvard School of Public Health, Boston, Massachusetts 02115, USA.
J Biol Chem. 1997 May 16;272(20):13426-31. doi: 10.1074/jbc.272.20.13426.
The basic helix-loop-helix (bHLH) transcription factors E12 and E47 regulate cellular differentiation and proliferation in diverse cell types. While looking for proteins that bind to E12 and E47 by the yeast interaction trap, we isolated the rat (r) homologue of the human (h) polymyositis-scleroderma autoantigen (PM-Scl), which has been localized to the granular layer of the nucleolus and to distinct nucleocytoplasmic foci. The rPM-Scl and hPM-Scl homologues are 96% similar and 91% identical. We found that rPM-Scl mRNA expression was regulated by growth factor stimulation in cultured rat aortic smooth muscle cells. rPM-Scl bound to E12 and E47 but not to Id3, Gax, Myb, OCT-1, or Max. The C terminus of rPM-Scl (amino acids 283-353) interacted specifically with a 54-amino acid domain in E12 that is distinct from the bHLH domain. Finally, cotransfection of rPM-Scl and E47 specifically increased the promoter activity of a luciferase reporter construct containing an E box and did not affect the basal activity of the reporter construct. rPM-Scl appears to be a novel non-HLH-interacting partner of E12/E47 that regulates E2A protein transcription.
基本螺旋-环-螺旋(bHLH)转录因子E12和E47在多种细胞类型中调节细胞分化和增殖。在通过酵母相互作用陷阱寻找与E12和E47结合的蛋白质时,我们分离出了人类(h)多发性肌炎-硬皮病自身抗原(PM-Scl)的大鼠(r)同源物,该抗原已定位到核仁的颗粒层和不同的核质焦点。rPM-Scl和hPM-Scl同源物的相似度为96%,一致性为91%。我们发现,在培养的大鼠主动脉平滑肌细胞中,rPM-Scl mRNA的表达受生长因子刺激的调节。rPM-Scl与E12和E47结合,但不与Id3、Gax、Myb、OCT-1或Max结合。rPM-Scl的C末端(氨基酸283 - 353)与E12中一个不同于bHLH结构域的54个氨基酸的结构域特异性相互作用。最后,rPM-Scl和E47的共转染特异性增加了含有E盒的荧光素酶报告基因构建体的启动子活性,且不影响报告基因构建体的基础活性。rPM-Scl似乎是E12/E47的一种新型非HLH相互作用伴侣,可调节E2A蛋白转录。