Simonson M S, Rooney A, Herman W H
Department of Medicine, School of Medicine, Case Western Reserve University, Cleveland, OH 44106.
Nucleic Acids Res. 1993 Dec 11;21(24):5767-74. doi: 10.1093/nar/21.24.5767.
Id is a family of dominant negative helix-loop-helix (HLH) proteins that block cell-specific transcription mediated by basic HLH (bHLH) transcription mediated by basic HLH (bHLH) transcription factors. We have analyzed Id1 expression in mesangial cells as a first step towards understanding the putative role of bHLH transcription factors in cell type-specific gene expression in the kidney. Glomerular mesangial cells expressed an abundant 1.1 kb mRNA transcript for Id1, but in contrast to other cell types Id1 mRNA was expressed in both randomly cycling cells and in serum-deprived, quiescent cultures. When quiescent mesangial cells were treated with serum to re-enter G1, Id1 mRNA levels were rapidly (2-4 h) and transiently down-regulated. Down-regulation of Id1 mRNA following addition of serum to mesangial cells was cell type-specific and contrasted with induction of Id1 by serum in BHK-21 and 3T3 fibroblasts. Down-regulation of Id1 mRNA correlated with mitogenesis and occurred when quiescent cells were treated with growth factors that activate G protein-coupled receptors and receptor protein tyrosine kinases but not with a non-mitogenic cAMP analog. Down-regulation of Id1 by growth factors required de novo protein synthesis, suggesting that a labile protein was involved. Appearance of E-box DNA binding activity in mesangial cell extracts followed down-regulation of Id1 message. Steady state Id1 mRNA levels and E-box DNA binding activity were not tightly correlated, suggesting complex regulation of Id1 activity. mRNA transcripts for E2A gene products were also expressed in mesangial cells, but these cells failed to express mRNAs for MyoA/MyoD-related genes. Collectively, these data demonstrate that Id1 is expressed in renal mesangial cells and suggest that bHLH complexes might be important for transcriptional regulation in the kidney. In addition, the observation that Id1 mRNA is transiently down-regulated by serum in mesangial cells suggests that Id1 gene expression is more complicated than previously appreciated and is tightly regulated in a cell-specific manner.
Id是一类显性负性螺旋-环-螺旋(HLH)蛋白家族,可阻断由碱性HLH(bHLH)转录因子介导的细胞特异性转录。作为了解bHLH转录因子在肾脏细胞类型特异性基因表达中假定作用的第一步,我们分析了系膜细胞中Id1的表达。肾小球系膜细胞表达一种丰富的1.1 kb Id1 mRNA转录本,但与其他细胞类型不同的是,Id1 mRNA在随机循环的细胞以及血清饥饿的静止培养细胞中均有表达。当静止的系膜细胞用血清处理以重新进入G1期时,Id1 mRNA水平迅速(2 - 4小时)且短暂下调。向系膜细胞中添加血清后Id1 mRNA的下调具有细胞类型特异性,这与血清在BHK - 21和3T3成纤维细胞中诱导Id1形成对比。Id1 mRNA的下调与有丝分裂相关,当静止细胞用激活G蛋白偶联受体和受体蛋白酪氨酸激酶的生长因子处理时会发生下调,但用非促有丝分裂的cAMP类似物处理则不会。生长因子对Id1的下调需要从头合成蛋白质,这表明涉及一种不稳定的蛋白质。Id1信息下调后,系膜细胞提取物中出现E - 盒DNA结合活性。稳态Id1 mRNA水平与E - 盒DNA结合活性并非紧密相关,这表明Id1活性存在复杂的调控。E2A基因产物的mRNA转录本也在系膜细胞中表达,但这些细胞未能表达MyoA/MyoD相关基因的mRNA。总体而言,这些数据表明Id1在肾系膜细胞中表达,并提示bHLH复合物可能对肾脏中的转录调控很重要。此外,系膜细胞中血清可使Id1 mRNA短暂下调这一观察结果表明,Id1基因表达比之前认为的更为复杂,且以细胞特异性方式受到严格调控。