GENOMICS 40, 242 – 246 (1997) ARTICLE NO. GE964575 Cloning, Structural Organization, and Chromosomal Mapping of the Human Phenol Sulfotransferase STP2 Gene ANDREA GAEDIGK,B ARBARA G. BEATTY,* AND DENIS M. GRANT 1 Division of Clinical Pharmacology & Toxicology, and * CGAT FISH Mapping Resource Centre, Division of Molecular Pathology, Research Institute, The Hospital for Sick Children, 555 University Avenue, Toronto, Ontario, M5G 1X8, Canada Received May 24, 1996; accepted December 17, 1996 codynamics by conjugating oxygen- or nitrogen-con- Phenol- and monoamine-metabolizing sulfotransfer- taining functional groups with sulfate, leading either to ases (STP and STM, respectively) are members of a direct termination of drug action or to enhanced water superfamily of enzymes that add sulfate to a variety solubility that accelerates excretion from the body (Ja- of xenobiotics and endobiotics containing hydroxyl or koby et al., 1980; Mulder and Jakoby, 1990; Weinshil- amino functional groups. To characterize related sul- boum and Aksoy, 1994). However, sulfation may also fotransferase genes further, we used extra-long PCR lead to the ‘‘metabolic activation’’ of certain xenobiotics (XL-PCR) to generate three distinct sizes of amplifica- to chemically unstable sulfonyloxy esters that decom- tion products from human genomic DNA or from geno- pose to electrophilic nitrenium or carbonium ions with mic phage library clones, each of which contained sul- the potential to bind covalently to intracellular nucleo- fotransferase gene sequences. One of the PCR frag- philes such as DNA and proteins and lead to cytotoxic- ments contained a new sulfotransferase gene, STP2, ity or genotoxicity. corresponding to a recently published cDNA clone that The liver cDNAs encoding several cytosolic sulfotran- encodes a sulfotransferase with catalytic specificity sferases have been cloned, and the catalytic properties distinct from that of the previously described STP1 of their expressed products have been compared with and STM. Additional upstream sequence information those of enzymes purified using classical biochemical was obtained using a second STP2-specific XL-PCR- techniques. These include the thermostable phenol-sul- based approach. The STP2 gene is composed of eight exons and seven introns, with exon sizes ranging from fating STP1 (also termed TS-PST, P-PST, or HAST1) 95 to 181 bp. Protein-coding exon lengths and locations (Wilborn et al., 1992; Zhu et al., 1993; Veronese et al., of the splice junctions were identical to those in both 1994), the more thermolabile monoamine-sulfating the STM gene and an STP2 gene published indepen- STM (TL-PST, M-PST, or HAST3) (Zhu et al., 1993; dently by another group recently. The STP2 gene maps Veronese et al., 1994; Wood et al., 1994), dehydroepian- to a chromosomal location (16p11.2– p12) that is the drosterone sulfotransferase (DHEA-ST) (Falany et al., same as that previously determined for both STP1 and 1989; Comer et al., 1993), and estrogen sulfotransfer- STM. The characterization of the STP2 gene provides ase (EST) (Aksoy et al., 1994b; Falany et al., 1995). further insight into the organization, regulation, and Cloning of sulfotransferase cDNAs from other tissues multiplicity of the sulfotransferase supergene family. such as brain and platelets has revealed additional iso-  1997 Academic Press forms and allelic variants (Hwang et al., 1995; Jones et al., 1995). Sequences of the genes encoding STM (Aksoy and Weinshilboum, 1995), DHEA-ST (Luu-The et al., INTRODUCTION 1995; Otterness et al., 1995), EST (Her et al., 1995), and part of STP1 (Dooley et al., 1993) have also been Sulfotransferases are members of an enzyme super- reported, and the STP1 and STM genes have been family that catalyze the sulfation of a variety of endoge- mapped to human chromosome 16p12.1 – p11.2 (Dooley nous and exogenous compounds, including steroids, et al., 1993) and 16p11.2 (Aksoy et al., 1994a), respec- neurotransmitters, bile acids, and biogenic amines, as tively. More recently, cDNA clones encoding a novel well as a large number of therapeutically used drugs sulfotransferase, STP2 (also termed ST1A2 or HAST4), and environmental chemicals. In the latter regard, and a variant form, HAST4v, which metabolize p-ni- they play important roles in modulating drug pharma- trophenol but not dopamine, were isolated from human brain and liver cDNA libraries (Ozawa et al., 1995; Zhu Sequence data from this article have been deposited with the Gen- et al., 1996). Bank/EMBL Data Libraries under Accession No. U33886. To gain further insight into sulfotransferase gene 1 To whom correspondence should be addressed. Telephone: (416) 813-5175. Fax: (416) 813-7562. E-mail: grant@sickkids.on.ca. organization, regulation, and multiplicity, we report 242 0888-7543/97 $25.00 Copyright  1997 by Academic Press All rights of reproduction in any form reserved.