Zhou Y, Sun Z, Means A R, Sassone-Corsi P, Bernstein K E
Department of Pathology and Laboratory Medicine, Emory University, Atlanta, GA 30322, USA.
Proc Natl Acad Sci U S A. 1996 Oct 29;93(22):12262-6. doi: 10.1073/pnas.93.22.12262.
Testis angiotensin-converting enzyme (ACE) is a unique form of ACE, only produced by male germ cells, and results from a testis-specific promoter found within the ACE gene. We have investigated the role of cAMP-response element modulator (CREM)tau in testis ACE transcription. In gel shift experiments, testes nuclear proteins retard an oligonucleotide containing the cAMP-response element (CRE) found at position -55 in the testis ACE promoter. Anti-CREM antibody supershifts this complex. Competitive gel shift shows that recombinant CREM tau protein and testis nuclear proteins have a similar specificity of binding to the tests ACE CRE. Functional analysis using in vitro transcription and transfection studies also demonstrate that CREM tau protein is a transcriptional activator of the testis ACE promoter. Western blot analysis identifies CREM tau protein in the protein-DNA complex formed between nuclear proteins and the testis ACE CRE motif. This analysis also identified other CREM isoforms in the gel-shifted complex, which are thought to be CREM tau 1/2, CREM alpha/beta, and S-CREM. These data indicate that CREM tau isoforms play an important role as a positive regulator in the tissue-specific expression of testis ACE.
睾丸血管紧张素转换酶(ACE)是ACE的一种独特形式,仅由雄性生殖细胞产生,它源于在ACE基因中发现的睾丸特异性启动子。我们研究了环磷酸腺苷反应元件调节剂(CREM)tau在睾丸ACE转录中的作用。在凝胶迁移实验中,睾丸核蛋白使包含位于睾丸ACE启动子-55位置的环磷酸腺苷反应元件(CRE)的寡核苷酸迁移受阻。抗CREM抗体使这种复合物发生超迁移。竞争性凝胶迁移表明,重组CREM tau蛋白和睾丸核蛋白与睾丸ACE CRE具有相似的结合特异性。使用体外转录和转染研究进行的功能分析也表明,CREM tau蛋白是睾丸ACE启动子的转录激活因子。蛋白质印迹分析在核蛋白与睾丸ACE CRE基序形成的蛋白质-DNA复合物中鉴定出CREM tau蛋白。该分析还在凝胶迁移复合物中鉴定出其他CREM异构体,它们被认为是CREM tau 1/2、CREMα/β和S-CREM。这些数据表明,CREM tau异构体作为睾丸ACE组织特异性表达的正调节因子发挥重要作用。