Transcriptional regulation of human 3'-phosphoadenosine 5'-phosphosulphate synthase 2.
Transcriptional regulation of human 3'-phosphoadenosine 5'-phosphosulphate synthase 2.
复制标题
人 3-磷酸腺苷 5-磷酸硫酸合酶 2 的转录调控。
DOI：
10.1042/0264-6021:3630263
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发表时间：
2002
期刊：
影响因子：
--
通讯作者：
Strott,CharlesA
中科院分区：
文献类型：
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作者：
Shimizu,Chikara;Fuda,Hirotoshi;Lee,YoungC;Strott,CharlesA
磺化是一个基本过程，对于正常生长和发育以及维持内部环境至关重要。对于所有磺基缀合反应必不可少的通用磺酸盐供体分子是腺苷3′-磷酸5′-磷酸硫酸盐（PAPS），其通过双功能PAPS合酶的作用由ATP和无机硫酸盐产生。存在由位于染色体4（PAPS合成酶1）和染色体10（PAPS合成酶2）上的基因编码的两种同工酶。PAPS合成酶2的启动子既不包含TATAAA也不包含CCAAT盒，尽管存在共有起始基序。在用与报告基因融合的PAPS合酶2基因的不同长度的5′-侧翼区转染后，使用三种人细胞系来检查启动子活性。近端启动子活性位于假定的转录起始位点上游bp-84和bp-124之间。该区域含有两个GC/GT盒，其对于完整启动子活性是必需的，如缺失分析所示，并进一步得到诱变的支持。高表达PAPS合酶2的SW 13细胞的核提取物含有与具有启动子特异性GC/GT盒的探针结合的蛋白质。此外，特异性蛋白（Sp）1，Sp2和Sp3蛋白在核提取物中的存在被证实的超位移分析。使用SL 2细胞的共转染实验产生了额外的支持参与Sp1的PAPS合酶2基因的转录调控;参与Sp2和/或Sp3仍有待进一步澄清。
Sulphonation is a fundamental process that is essential for normal growth and development as well as maintenance of the internal milieu. The universal sulphonate donor molecule essential for all sulphoconjugation reactions is adenosine 3′-phosphate 5′-phosphosulphate (PAPS), which is produced from ATP and inorganic sulphate by the action of bifunctional PAPS synthase. There are two isozymes encoded by genes located on chromosome 4 (PAPS synthase 1) and chromosome 10 (PAPS synthase 2). The promoter for PAPS synthase 2 contains neither a TATAAA nor a CCAAT box, although a consensus initiator motif is present. Three human cell lines were used to examine promoter activity after transfection with various lengths of the 5′-flanking region of the PAPS synthase 2 gene fused to a reporter gene. Proximal promoter activity was located between bp −84 and bp −124 upstream of the purported transcription start site. This region contains two GC/GT boxes that are essential for full promoter activity, as indicated by deletion analysis and supported further by mutagenesis. A nuclear extract of SW13 cells, which highly express PAPS synthase 2, contained proteins that bound to probes possessing promoter-specific GC/GT boxes. Furthermore, the presence of specificity protein (Sp) 1, Sp2 and Sp3 proteins in the nuclear extract was confirmed by supershift analysis. Co-transfection experiments using SL2 cells yielded additional support for the involvement of Sp1 in transcriptional regulation of the PAPS synthase 2 gene; the involvement of Sp2 and/or Sp3 remains to be clarified further.