Pulkka A, Ihalainen R, Aatsinki J, Pajunen A
Biocenter, University of Oulu, Finland.
FEBS Lett. 1991 Oct 21;291(2):289-95. doi: 10.1016/0014-5793(91)81304-q.
The gene for S-adenosylmethionine decarboxylase (AdoMetDC) was isolated from a rat genomic library using AdoMetDC cDNA as a probe. Nucleotide sequence analysis shows that the rat AdoMetDC gene consists of 8 exons which encode a protein identical to that inferred by a rat AdoMetDC cDNA sequence. The exons range in length from 43 to 1964 base pairs spanning 15672 bases of chromosomal DNA. All of the exon/intron junctions were found to conform to the consensus splice donor and acceptor sequences. Exon 8 corresponds to the 3' noncoding region of the 2 species of AdoMetDC mRNA which are formed by alternative utilization of 2 polyadenylation signals separated from each other by 1272 nucleotides. The transcription initiation site was located by S1 nuclease protection and by primer extension analysis, -325 nucleotides upstream of the translation initiation codon. The promoter region of the rat AdoMetDC gene contains a TATA box at -29 base pairs. No typical GC or CAAT boxes are located in the promoter, but six GC boxes and several putative binding sites for both tissue-specific and non-specific transcription factors are found in the proximal part of intron 1. Southern blot analyses reveal a complex hybridization pattern suggesting that there are multiple copies of the AdoMetDC gene in the rat genome.
以S - 腺苷甲硫氨酸脱羧酶(AdoMetDC)cDNA为探针,从大鼠基因组文库中分离出AdoMetDC基因。核苷酸序列分析表明,大鼠AdoMetDC基因由8个外显子组成,这些外显子编码的蛋白质与大鼠AdoMetDC cDNA序列推导的蛋白质相同。外显子长度从43到1964个碱基对不等,跨越15672个碱基对的染色体DNA。所有外显子/内含子连接均符合共有剪接供体和受体序列。外显子8对应于两种AdoMetDC mRNA的3'非编码区,这两种mRNA是通过交替利用两个彼此相隔1272个核苷酸的聚腺苷酸化信号形成的。通过S1核酸酶保护和引物延伸分析确定转录起始位点位于翻译起始密码子上游-325个核苷酸处。大鼠AdoMetDC基因的启动子区域在-29个碱基对处含有一个TATA框。启动子中未发现典型的GC或CAAT框,但在内含子1的近端发现了六个GC框以及几个组织特异性和非特异性转录因子的假定结合位点。Southern印迹分析揭示了复杂的杂交模式,表明大鼠基因组中存在多个AdoMetDC基因拷贝。