Xu Z H, Otterness D M, Freimuth R R, Carlini E J, Wood T C, Mitchell S, Moon E, Kim U J, Xu J P, Siciliano M J, Weinshilboum R M
Department of Molecular Pharmacology and Experimental Therapeutics, Mayo Medical School/Mayo Graduate School/Mayo Clinic, Rochester, Minnesota 55905, USA.
Biochem Biophys Res Commun. 2000 Feb 16;268(2):437-44. doi: 10.1006/bbrc.2000.2123.
Sulfae conjugation is an important pathway in the metabolism of a large number of exogenous and endogenous compounds. These reactions are catalyzed by sulfotransferase (SULT) enzymes that utilize 3'-phosphoadenosine 5'-phosphosulfate (PAPS) as a sulfate donor. PAPS is synthesized from ATP and inorganic sulfate by PAPS synthetase (PAPSS). Two separate PAPSS cDNAs, PAPSS1 and PAPSS2, have been identified in human tissues. We have cloned and characterized the genes for human PAPSS1 and PAPSS2 to make it possible to study the pharmacogenomics of these enzymes. Both genes consisted of 12 exons with virtually identical exon-intron splice junction locations. All splice junctions conformed to the "GT-AG" rule. The total length of PAPSS1 was approximately 108 kb, while that of PAPSS2 was greater than 37 kb. The 5'-flanking region of PAPSS1 did not include a TATA box sequence near the site of transcription initiation, but PAPSS2 had a TATA motif located 21 bp upstream from the site of transcription initiation. Northern blot analysis showed that the major PAPSS1 and PAPSS2 transcripts were approximately 2.7 and 4.2 kb in length, respectively. PAPSS1 mapped to human chromosome band 4q24 while PAPSS2 mapped to 10q22-23 by fluorescence in situ hybridization analysis. Cloning and structural characterization of PAPSS1 and PAPSS2 will make it possible to perform molecular genetic and pharmacogenomic studies of these important enzymes in humans.
硫酸结合作用是大量外源性和内源性化合物代谢的一条重要途径。这些反应由磺基转移酶（SULT）催化，该酶利用3'-磷酸腺苷5'-磷酸硫酸酯（PAPS）作为硫酸供体。PAPS由PAPS合成酶（PAPSS）从ATP和无机硫酸盐合成。在人体组织中已鉴定出两个独立的PAPSS cDNA，即PAPSS1和PAPSS2。我们已经克隆并表征了人类PAPSS1和PAPSS2的基因，以便能够研究这些酶的药物基因组学。两个基因均由12个外显子组成，外显子-内含子剪接连接位置几乎相同。所有剪接连接均符合“GT-AG”规则。PAPSS1的总长度约为108 kb，而PAPSS2的总长度大于37 kb。PAPSS1的5'-侧翼区域在转录起始位点附近不包括TATA盒序列，但PAPSS2在转录起始位点上游21 bp处有一个TATA基序。Northern印迹分析表明，主要的PAPSS1和PAPSS2转录本长度分别约为2.7和4.2 kb。通过荧光原位杂交分析，PAPSS1定位于人类染色体4q24带，而PAPSS2定位于10q22-23。PAPSS1和PAPSS2的克隆和结构表征将使对人类这些重要酶进行分子遗传学和药物基因组学研究成为可能。
