Marić S C, Crozat A, Jänne O A
Population Council, New York, New York.
J Biol Chem. 1992 Sep 15;267(26):18915-23.
Genomic clones for the S-adenosylmethionine (AdoMet) decarboxylase gene were isolated from a human chromosome 6 DNA library. In addition, polymerase chain reaction and specific primers were used to amplify fragments from chromosomal DNA covering exonic regions not found in the screening of DNA libraries with AdoMet decarboxylase cDNA. The gene encompasses at least 22 kilobases of chromosome 6 DNA and comprises nine exons and eight introns, in contrast to the corresponding rat gene that has only eight exons (Pulkka, A., Ihalainen, R., Aatsinki, J., and Pajunen, A. (1991) FEBS Lett. 291, 289-295). Exon-intron junctions in the human and rat AdoMet decarboxylase genes were in identical positions except that exons 6 and 7 of the human gene formed a single exon in the rat gene. Alu-like sequences are present in four introns and the 5'-flanking region of the human gene. The promoter region contains a TATA box adjacent to the cap site; in addition, DNA elements for binding of transcription factors AP-1, AP-2, CREB, SP-1, and multiple steroid receptors are present between position -3,158 and the transcription start site. Two AdoMet decarboxylase promoter-reporter gene constructs with about 170 and 1,500 nucleotides of the 5'-flanking DNA were used in transient expression studies. AdoMet decarboxylase promoter was capable of driving reporter gene expression, but it was less active than the murine ornithine decarboxylase promoter. There are at least three potential polyadenylation signals at the 3'-end of the gene, and utilization of the first two results in the formation of the 2.0- and 3.6-kilobase AdoMet decarboxylase mRNA species present in human tissues and cell lines. AdoMet decarboxylase gene-related sequences were also present in a human X chromosome-specific DNA library. Partial nucleotide sequencing of this DNA revealed a lack of introns present in the gene located on chromosome 6, suggesting that the locus on the X chromosome contains a processed AdoMet decarboxylase pseudogene.
从人6号染色体DNA文库中分离出S-腺苷甲硫氨酸(AdoMet)脱羧酶基因的基因组克隆。此外,使用聚合酶链反应和特异性引物从染色体DNA中扩增片段,这些片段覆盖了在用AdoMet脱羧酶cDNA筛选DNA文库时未发现的外显子区域。该基因包含至少22千碱基对的6号染色体DNA,由9个外显子和8个内含子组成,而相应的大鼠基因只有8个外显子(Pulkka, A., Ihalainen, R., Aatsinki, J., and Pajunen, A. (1991) FEBS Lett. 291, 289 - 295)。人和大鼠AdoMet脱羧酶基因的外显子 - 内含子连接位点处于相同位置,只是人基因的外显子6和7在大鼠基因中形成了一个单一外显子。人基因的4个内含子和5'侧翼区域存在类似Alu的序列。启动子区域在帽位点附近有一个TATA盒;此外,在 - 3158位和转录起始位点之间存在转录因子AP - 1、AP - 2、CREB、SP - 1和多种类固醇受体结合的DNA元件。两个带有约170和1500个核苷酸5'侧翼DNA的AdoMet脱羧酶启动子 - 报告基因构建体用于瞬时表达研究。AdoMet脱羧酶启动子能够驱动报告基因表达,但活性低于小鼠鸟氨酸脱羧酶启动子。在该基因的3'端至少有三个潜在的聚腺苷酸化信号,前两个信号的利用导致在人组织和细胞系中出现2.0和3.6千碱基的AdoMet脱羧酶mRNA种类。AdoMet脱羧酶基因相关序列也存在于一个人X染色体特异性DNA文库中。对该DNA的部分核苷酸测序显示,位于6号染色体上的基因中不存在内含子,这表明X染色体上的位点包含一个加工后的AdoMet脱羧酶假基因。