Yokoyama Y, Kawamoto T, Mitsuuchi Y, Kurosaki T, Toda K, Ushiro H, Terashima M, Sumimoto H, Kuribayashi I, Yamamoto Y
Department of Medical Chemistry, Kochi Medical School, Japan.
Eur J Biochem. 1990 Dec 12;194(2):521-6. doi: 10.1111/j.1432-1033.1990.tb15647.x.
The promoter region of the poly(ADP-ribose) polymerase gene has been isolated using a Sau3AI genomic library derived from human leukocyte. It lacks typical transcriptional regulatory elements such as TATA and CAAT boxes, but it contains two potential Sp1 binding sites and three putative AP-2 binding elements. The region up to nucleotide position-99 in relation to the predominant transcriptional initiation site exhibits promoter activity as judged by chloramphenicol acetyltransferase assay and the activity is enhanced both by cAMP and by phorbol ester. Northern blot and Western blot analyses have revealed that expression of the polymerase gene is also stimulated by both of these compounds in cultured HeLa cells. Southern blot hybridization of genomic DNA separately digested with various endonucleases gives a discrete single band in each case when the 5'-untranslated region of the polymerase cDNA is used as a probe. These results indicate that poly(ADP-ribose) polymerase is encoded by a unique gene whose expression is regulable by cAMP and by phorbol ester.
利用源自人白细胞的Sau3AI基因组文库分离出了聚（ADP - 核糖）聚合酶基因的启动子区域。它缺乏典型的转录调控元件，如TATA盒和CAAT盒，但含有两个潜在的Sp1结合位点和三个假定的AP - 2结合元件。相对于主要转录起始位点，直至核苷酸位置 - 99的区域通过氯霉素乙酰转移酶测定显示出启动子活性，并且该活性可被cAMP和佛波酯增强。Northern印迹和Western印迹分析表明，在培养的HeLa细胞中，这两种化合物也能刺激聚合酶基因的表达。当用聚合酶cDNA的5' - 非翻译区作为探针时，用各种核酸内切酶分别消化的基因组DNA的Southern印迹杂交在每种情况下都给出一条离散的单带。这些结果表明，聚（ADP - 核糖）聚合酶由一个独特的基因编码，其表达可被cAMP和佛波酯调节。
