Hopitzan A, Himmelbauer H, Spevak W, Castanon M J
Research and Development, Boehringer Ingelheim Austria, Vienna, 1121, Austria.
Genomics. 2000 Jun 15;66(3):313-23. doi: 10.1006/geno.2000.6217.
We have isolated and functionally characterized the mouse gene for the C2 subunit of the 20S proteasome. The gene contains 10 exons distributed over a region of 12 kb on the distal end of mouse chromosome 7. Its exon-intron structure differs from those of the other few known proteasome genes. Transfection assays revealed that 1.5 kb of 5' flanking sequence is active as promoter in cultured myoblasts. Deletion reporter constructs narrowed this presumptive promoter region to within 450 bp upstream of the translation initiation site. Several consensus motifs for transcription factor binding sites were identified in this upstream region of the gene. Psma1 was mapped to mouse chromosome 7 using the interspecific backcross DNA panels from The Jackson Laboratory. Additional mapping studies showed that the mouse genes Psma1 and Pde3b are closely linked, residing between cM 53 and 53.3 in a region syntenic to human chromosome 11p15. Our results extend the structural and functional analysis of genes encoding the 20S proteasome subunits and provide the basis for the study of their regulation.
我们已经分离出小鼠20S蛋白酶体C2亚基的基因并对其进行了功能表征。该基因包含10个外显子,分布在小鼠7号染色体远端12kb的区域。其外显子-内含子结构与其他少数已知的蛋白酶体基因不同。转染实验表明,1.5kb的5'侧翼序列在培养的成肌细胞中作为启动子具有活性。缺失报告基因构建体将这个假定的启动子区域缩小到翻译起始位点上游450bp以内。在该基因的上游区域鉴定出了几个转录因子结合位点的共有基序。使用来自杰克逊实验室的种间回交DNA面板将Psma1定位到小鼠7号染色体上。进一步的定位研究表明,小鼠基因Psma1和Pde3b紧密连锁,位于与人类染色体11p15同源的区域,在cM 53和53.3之间。我们的结果扩展了对编码20S蛋白酶体亚基的基因的结构和功能分析,并为研究它们的调控提供了基础。