Kumar A, Reddy E P
Fels Institute for Cancer Research and Molecular Biology, Temple University School of Medicine, 3307 N. Broad Street, Philadelphia, PA 19140, USA.
Gene. 2001 May 30;270(1-2):221-9. doi: 10.1016/s0378-1119(01)00473-5.
In this study, we have isolated the genomic clone of the murine GSTM2 gene and determined its sequence. Consistent with the class mu genefamily, the mGSTM2 gene consists of eight exons. The exon-intron boundaries and the distribution of coding sequences within the exons of the known GST class mu family members were found to follow a similar pattern suggesting that various members of this family have originated from a single primordial gene by duplication and the structure has been closely maintained through evolution. By primer extension, the start of transcription was determined to be 40 bp upstream of the initial AUG codon. To gain an understanding of the mGSTM2 regulation, we have also cloned and analyzed its promoter region. Maximal activity was observed in a 170 bp 5'-flanking region. The activity was decreased by 3-fold in a 402 bp 5'-flanking region suggesting the presence of repressor elements. While no TATA box was identified, the presence of an SP1 site at position -38 was noted. Deletion of this SP1 site completely abrogated promoter activity. The promoter contained eight putative Myb responsive elements and its transcriptional activity was upregulated by t-Myb but not by c-Myb.
在本研究中,我们分离出了小鼠GSTM2基因的基因组克隆并测定了其序列。与μ类基因家族一致,mGSTM2基因由八个外显子组成。已知GSTμ类家族成员的外显子-内含子边界以及外显子内编码序列的分布遵循相似模式,这表明该家族的各个成员通过复制起源于单个原始基因,并且其结构在进化过程中得到了紧密维持。通过引物延伸,确定转录起始位点在起始AUG密码子上游40 bp处。为了了解mGSTM2的调控机制,我们还克隆并分析了其启动子区域。在一个170 bp的5'侧翼区域观察到最大活性。在一个402 bp的5'侧翼区域,活性降低了3倍,这表明存在抑制元件。虽然未鉴定到TATA盒,但在-38位发现了一个SP1位点。删除该SP1位点完全消除了启动子活性。该启动子包含八个假定的Myb反应元件,其转录活性被t-Myb上调,但不被c-Myb上调。