Bernier F, Soucy P, Luu-The V
Medical Research Council Group in Molecular Endocrinology, CHUL Research Center, Quebec, Canada.
DNA Cell Biol. 1996 May;15(5):367-75. doi: 10.1089/dna.1996.15.367.
Phenol sulfotransferases catalyze the transfer of a sulfonate moiety from 3'-phosphoadenosyl 5'-phosphosulfate to a phenolic group of lipophylic substrates to generate soluble sulfate esters. Using a phenol sulfotransferase cDNA as probe to screen a human leukocyte genomic DNA library constructed in lambda EMBL3, we obtained a clone containing a complete gene sequence. Comparison of the gene sequence with that of the corresponding cDNAs, namely phenol-sulfating phenol sulfotransferase (P-PST) or thermostable sulfotransferase (TS-PST), and human aryl sulfotransferase 1 and 2 (HAST1 and HAST2) indicates that the gene possesses eight short exons separated by seven introns included in approximately 5 kb. HAST2 has a different 5' untranslated sequence, and thus is encoded by a different mRNA species. While the nucleotide sequence corresponding to the 5' noncoding region of P-PST (TS-PST and HAST1) is included in the exon I, the 5' untranslated sequence of HAST2 is located in the beginning of exon IIa. The remaining sequence in exon II that is identical to both P-PST and HAST2 was termed exon IIb. Exons III to VIII, which cover the coding region and the 3' untranslated region, are almost identical in all types of PST or AST cDNAs. These results suggest that the phenol sulfotransferase gene possesses two alternate promoters that drive the expression of the two different mRNA species in a tissue-specific manner. Transfection of chloramphenicol acetyl transferase (CAT) reporter gene vectors containing the 5'-flanking sequence upstream from exon I and exon II, respectively, in transformed human embryonal kidney (293) cells indicate that both sequences possess promoter activity with higher activity for promoter 1. RNA blot analysis indicates that human phenol sulfotransferase gene is expressed in kidney, liver, lung, leukocyte, colon, small intestine, and spleen.
酚磺基转移酶催化磺酸根基团从3'-磷酸腺苷5'-磷酸硫酸转移至亲脂性底物的酚基团上,生成可溶性硫酸酯。我们以酚磺基转移酶cDNA为探针,筛选构建于λEMBL3中的人白细胞基因组DNA文库,获得了一个包含完整基因序列的克隆。将该基因序列与相应cDNA的序列进行比较,即酚硫酸化酚磺基转移酶(P-PST)或耐热磺基转移酶(TS-PST),以及人芳基磺基转移酶1和2(HAST1和HAST2),结果表明该基因有8个短外显子,被7个内含子隔开,约5 kb。HAST2有不同的5'非翻译序列,因此由不同的mRNA种类编码。虽然与P-PST(TS-PST和HAST1)的5'非编码区对应的核苷酸序列包含在外显子I中,但HAST2的5'非翻译序列位于外显子IIa的起始处。外显子II中与P-PST和HAST2都相同的其余序列被称为外显子IIb。覆盖编码区和3'非翻译区的外显子III至VIII在所有类型的PST或AST cDNA中几乎相同。这些结果表明,酚磺基转移酶基因有两个交替启动子,以组织特异性方式驱动两种不同mRNA种类的表达。分别在转化的人胚肾(293)细胞中,转染含有外显子I和外显子II上游5'侧翼序列的氯霉素乙酰转移酶(CAT)报告基因载体,结果表明这两个序列都具有启动子活性,启动子1的活性更高。RNA印迹分析表明,人酚磺基转移酶基因在肾脏、肝脏、肺、白细胞、结肠、小肠和脾脏中表达。