Watanabe K, Nomoto M, Nagata S, Itoh Y, Hikichi K, Maruyama N, Mita T, Senshu T
Department of Biochemical Engineering and Science, Kyushu Institute of Technology, Fukuoka, Japan.
Gene. 1992 May 15;114(2):261-5. doi: 10.1016/0378-1119(92)90585-d.
Genomic clones of the rat peptidylarginine deiminase (PAD)-encoding gene (PAD) were isolated, and the gene organization was analyzed by restriction mapping and nucleotide sequencing. The PAD spans more than 50 kb and contains 16 exons and 15 introns. The lengths of the introns from 0.5 kb to more than 16.5 kb. A 1.7-kb sequence in the 5'-flanking region was determined. S1 nuclease mapping revealed two putative cap sites 79 and 81 bp upstream from the N-terminal ATG codon of PAD, which had been determined by amino acid sequence analysis. This ATG was confirmed to be the translation start site, since no other ATG codon was found in the open reading frame downstream from the cap sites. The 5'-flanking sequence contains four potential SP1-binding sites, a putative Pit-1/GHF-1-binding site, four short sequences either identical or homologous to the sequences in the promoter regions of rat or human growth hormone encoding genes, as well as a sequence similar to an estrogen-responsive element. However, neither a typical TATAA box, nor CCAAT box is present. These results provide important clues for elucidating the mechanism of female-specific and/or sex cycle-dependent gene expression.
分离出了大鼠肽基精氨酸脱亚氨酶（PAD）编码基因（PAD）的基因组克隆，并通过限制性图谱分析和核苷酸测序对基因结构进行了分析。PAD跨度超过50kb，包含16个外显子和15个内含子。内含子长度从0.5kb到超过16.5kb不等。测定了5'-侧翼区的一个1.7kb序列。S1核酸酶图谱分析显示，在通过氨基酸序列分析确定的PAD N端ATG密码子上游79和81bp处有两个假定的帽位点。该ATG被确认为翻译起始位点，因为在帽位点下游的开放阅读框中未发现其他ATG密码子。5'-侧翼序列包含四个潜在的SP1结合位点、一个假定的Pit-1/GHF-1结合位点、四个与大鼠或人类生长激素编码基因启动子区域中的序列相同或同源的短序列，以及一个与雌激素反应元件相似的序列。然而，既没有典型的TATAA盒，也没有CCAAT盒。这些结果为阐明雌性特异性和/或性周期依赖性基因表达的机制提供了重要线索。
