Human SULT1A genes: cloning and activity assays of the SULT1A promoters.
The three human SULT1A sulfotransferase enzymes are closely related in amino acid sequence (>90%), yet differ in their substrate preference and tissue distribution. SULT1A1 has a broad tissue distribution and metabolizes a range of xenobiotics as well as endogenous substrates such as estrogens and iodothyronines. While the localization of SULT1A2 is poorly understood, it has been shown to metabolize a number of aromatic amines. SULT1A3 is the major catecholamine sulfonating form, which is consistent with it being expressed principally in the gastrointestinal tract. SULT1A proteins are encoded by three separate genes, located in close proximity to each other on chromosome 16. The presence of differential 5'-untranslated regions identified upon cloning of the SULT1A cDNAs suggested the utilization of differential transcriptional start sites and/or differential splicing. This chapter describes the methods utilized by our laboratory to clone and assay the activity of the promoters flanking these different untranslated regions found on SULT1A genes. These techniques will assist investigators in further elucidating the differential mechanisms that control regulation of the human SULT1A genes. They will also help reveal how different cellular environments and polymorphisms affect the activity of SULT1A gene promoters.
Hempel
Nadine
N
School of Biomedical Science, Faculty of Biological and Chemical Sciences, University of Queensland, Brisbane, Australia.
Negishi
Masahiko
M
McManus
Michael E
ME
D000818:Animals; D015239:Arylsulfotransferase / Q000145:classification / Q000235:genetics*; D002460:Cell Line; D003001:Cloning, Molecular / Q000379:methods; D004330:Drosophila; D004789:Enzyme Activation; D015971:Gene Expression Regulation, Enzymologic / Q000235:genetics*; D006801:Humans; D011110:Polymorphism, Genetic; D011401:Promoter Regions, Genetic; D020033:Protein Isoforms / Q000235:genetics
D020033:Protein Isoforms; D015239:Arylsulfotransferase; C117544:SULT1A1 protein, human; C513290:SULT1A2 protein, human