GENOMICS

(1991)

11,309-316

Mapping JOHN S. MILES,**’

JULIE

Genes Encoding Drug-Metabolizing in Recombinant Inbred Mice

Enzymes

E. Moss,* BENJAMIN A. TAYLOR,t BRIAN BURCHELL,$ AND C. ROLAND WOLF***

*Imperial Cancer Research Fund, Laboratory of Molecular Pharmacology, University Department of Biochemistry, Hugh Robson Building, George Square, Edinburgh EHB 9XD, United Kingdom; t The Jackson Laboratory, Bar Harbor, Maine 04609; and *Department of Biochemical Medicine, Ninewells Hospital and Medical School, Dundee DO1 SW, United Kingdom Received

February

21, 1991;

INTRODUCTION

Protection from foreign chemical compounds is believed to be a major driving force in evolution, and several multigene families encoding enzymes capable of metabolizing xenobiotics are widespread in nature. These enzymes are divided into two groups; the Phase I enzymes are responsible for activating otherwise inert, lipophilic chemicals and the Phase II enzymes conjugate the activated compounds to hydrophilic groups, thus rendering them more soluble and accessible to further modification and excretion. The cytochrome P450-dependent monooxygenase (P450) system is by far the most important of the Phase I enzymes. It comprises many forms of cytochrome P45Os which couple with a single P450 reductase and are capable of inserting one atom from molecular oxygen into a wide range of compounds. University

May

8, 1991

The P45Os are encoded by a supergene family, genes from different families and subfamilies being scattered throughout mammalian genomes (Wolf, 1986; Gonzalez, 1989; Nebert et aZ., 1991). Ascertaining the chromosomal localization of these genes, particularly in the mouse, where analysis of recombinant inbred (RI) strains provides linkage data (Taylor, 1978, 1989), has been useful in helping to determine their function (Meehan et al., 1988; Miles et aZ., 1990). Genetic polymorphisms within the P450 system lead to the impaired metabolism of many drugs in human (see Franklin and Parke, 1986, for a review) and may be important in determining susceptibility to chemically induced carcinogenesis (Ayesh et aZ., 1984; Gough et uZ., 1990). The glutathione S-transferases (GSTs) are Phase II enzymes that catalyze the conjugation of glutathione to a wide range of electrophilic compounds and can also act as binding proteins and peroxidases (Mannervik and Danielson, 1988). They are classified into four groups using biochemical criteria (LY,I.L, z, and microsomal) corresponding to four distinct gene families, and where known, they have separate chromosomal locations. Genetic polymorphism within the GSTF genes in man has been implicated in determining susceptibility to lung cancer (Seidegard et al., 1986). The UDP-glucuronyltransferases (UDPGTs) are Phase II enzymes that catalyze the glucuronidation of various xenobiotic and endogenous compounds, glucuronide formation being a major pathway in their elimination (Dutton, 1980; Burchell, 1981; Tephly and Burchell, 1990). The UDPGTs make up a multigene family comprising several subfamilies, members of which are involved in the glucuronidation of androgens, bilirubin, phenols, morphine, etc. (Tephly and Burchell, 1990), there being genetic variation in the glucuronidation of bilirubin in human (Burchell and Coughtrie, 1989) and in the Gunn rat (Iyanagi et uZ., 1989).

Probes for cytochrome P450IVA (P450IVA), a- and ~-class glutathione S-transferases (GST), and phenol-metabolizing UDP-glucuronyltransferase (UDPGT-K29) detected restriction fragment length variants (RFLVs) between CS7BL/6J and DBAPJ mice. These variants were used to map the P45OIVA genes (Cyp4u) to chromosome 4, close to Mtv-13 and Pmu-19, midway between brown (b) and Gpd-1; GSTa! genes were mapped to chromosome 9, with a cross-hybridizing sequence mapping to another chromosome; the GSTr genes were mapped to the distal end of chromosome 1 near Pmv-21; one UDPGT-KS9 variant to chromosome 1, between Acrg and Emu-l 7, and another showed linkage to Ode10 on an unidentified chromosome. No RFLVs were detected with probes for PISOIID, P450 reductase, androsterone-metabolizing UDPGT, GSTr, or microsomal GST. Q lssl Academic POW, IM.

1 Present address: Department of Biochemistry, Glasgow, Glasgow G12 SQQ, UK. * To whom correspondence should be addressed.

revised

of

309

OSfsS-7543/91$3.00 Copyright 0 1991 hy Academic Press, Inc. All rights of reproduction in any form reserved.

310

MILES

Although single genetic factors play a role in susceptibility to chemical carcinogens, it is more likely that individual susceptibility is determined by interaction of several genetic and many environmental factors. Because of the wealth of genetic information available, the mouse is a good animal for studying these complex interactions. An understanding of the location, complexity, and variation of the genes encoding the drug-metabolizing enzymes is an essential prerequisite for understanding their contribution to chemical carcinogenesis. Restriction fragment length variants (RFLVs) detected with probes to such enzymes may be useful in the analysis of the genetic factors responsible, especially in model systems that are available for studying the inheritance of multigenically controlled traits (e.g., recombinant congenic strains; Demant and Hart, 1986). In this study we have used rat or human DNA probes for various members of the P450, GST, and UDPGT gene families to detect hybridizing DNA in the mouse. RFLVs were found for the P450IVA, GSTa, GSTx, and phenol-metabolizing UDPGT (UDPGT-K39) genes between C57BL/6J and DBA/ 25 mice. These variants were mapped using the BXD series of recombinant inbred strains. MATERIALS

Endonuclease

AL.

0.82-kb EcoRI-SstI fragment containing the 5’ half of the coding region of the UDPGT-R23 rat cDNA encoding an androsterone-metabolizing enzyme (Jackson and Burchell, 1986); (vi) full-length rat P450 reductase cDNA (Porter and Kasper, 1985); (vii) fulllength human P450IID cDNA (Gonzalez et al, 1988); (viii) full-length rat microsomal GST (S. Pemble and J. Taylor, personal communication; same sequence as that of DeJong et al., 1988); and (ix) a l.O-kb EcoRIPstI fragment of a human GSTp gene containing exons 3,4, and 5 (Taylor et al., 1991). Ail probes were radioactively labeled with [(w-32P]dCTP (3000 Ci mmol-‘; Amersham International) by random primer extension (Feinberg and Vogelstein, 1983). Labeled probes were hybridized to Southern blots as described by Hill et al. (1985). Blots were washed at 65°C in 1X SSC, 0.1% sodium pyrophosphate, 0.1% SDS for l-2 h with two changes of buffer and were autoradiographed at -70°C for 1-14 days using an intensifying screen and Kodak XAR-5 film. Filters were stripped of radioactivity by boiling in 0.1% sodium pyrophosphate, 2% SDS for 10 min and then exposed to X-ray film to confirm that the probe had been removed prior to reprobing.

Mapping of DNA Inbred Strains

AND METHODS

Source of DNA, Restriction and Gel Ekctrophoresis

ET

Digestion,

DNA from C57BL/6J, DBA/BJ, and the BXD recombinant inbred strains was isolated by standard methods from mice supplied by one of us (B.A.T.) or via Dr. G. Bulfield, AFRC Institute of Animal Physiology and Genetics (Edinburgh, UK). DNA (10 pg) was digested for 16 h with a fivefold excess of restriction endonuclease according to the manufacturers’ instructions, except that 2 n&f spermidine was included in the incubations. A further 10 U of enzyme was added and incubated for 2 h. Digested DNA was fractionated electrophoretically on agarose gels and was transferred to Hybond-N (Amersham International) as described by Southern (1975). The “kb ladder” (Gibco-BRL) was used for size markers.

Variants

in BXD Recombinant

The origin of the BXD RI strains and their use in mapping genetic variants have been reviewed (Taylor, 1978). The DNA variants described here were detected under the above conditions and scored as B (showing the C57BL/6J genotype) and D (showing the DBA/BJ genotype). Confidence limits for estimates of gene linkage were calculated by the method of Silver (1985).

Materials Restriction endonucleases and DNA polymerase I (Klenow fragment) were from Boehringer Ltd or Gibco-BRL. All other reagents were from the usual suppliers. RESULTS

Detection of RFLVs Radioactive DNA Probes and Hybridization The following DNA fragments were used as probes: (i) full-length rat P450IVA cDNA (Earnshaw et al., 1988); (ii) full-length human GSTx cDNA (Kano et al., 1987); (iii) full-length human GSTa cDNA (Lewis et al., 1988); (iv) O&3-kb PstI-BamHI fragment containing the 5’ half of the coding region of UDPGTK39 rat cDNA encoding a phenol-metabolizing enzyme (Harding and Burchell, unpublished data); (v)

Restriction fragment length variants that could be used for mapping the various genes with the BXD recombinant inbred strains were sought. DNA from C57BL/6J and DBA/BJ mice was digested with a series of different restriction enzymes, transferred onto Hybond-N nylon membranes, and hybridized to the cDNA probes under the conditions described under Materials and Methods. All probes detected cross-hybridizing DNA in both mouse strains. Probes for

DRUG-METABOLIZING

TABLE

ENZYMES

1

Restriction Fragment Length Polmorphisms Detected between C67BL/6 J and DBA/2 J Mice Probe” Enzyme BamHI BglII EcoRI Hue III Hind111 HpaI KpnI MspI NcoI PstI PVUII RsaI Sau3A SphI SstI stu1 TasI XbaI

P45OIV

GSTa

+ -

n -

+

n -

” + + + -

+ n -

n n + + + n n

+ -

GST?r + n + n + + + + n + n + + +

UDPGT-K39 + + + + + + + + + + +

Note. A + indicates a polymorphism detected with the enzyme and probe indicated, a - indicates no polymorphism detected, and an n indicates that the status was not determined. Probes for P450IID, P450 reductase, UDPTG-R23, GSTc(, and microsomal GST detected no polymorphisms with these enzymes. LI Probes are P45OIV, full-length rat P450IVA cDNA [Ref. (lo)]; GSTa, full-length GSTa cDNA [Ref. (30)]; GSTz, full-length human GSTr cDNA [Ref. (24)]; and UDPGT-K39, PstI-BamHI fragment containing the 5’ half of a cDNA encoding a phenol-metabolizing UDPGT (Harding and Burchell, unpublished data).

P45OIV, GSTa, GSTx, and UDPGT-KS9 detected RFLVs with several enzymes listed in Table 1, whereas probes for P450 reductase, P450IID, UDPGTR23, GSTg, and microsomal GST did not detect RFLVs with any of these enzymes and thus could not be used for mapping using the BXD RI strain approach. Several of the RFLVs were chosen for further analysis: P450IVA, EcoRI, HpaI, and S&I; GSTa, HindIII, and RsaI; GSTz, EcoRI, HindIII, K&I, and RsaI; and K39-GT, EcoRI, HindIII, KpnI, PstI, and StuI (Fig. 1).

Mapping

the P45OIVA

Variants

The full-length rat P450IVA cDNA probe detects up to 50 kb of hybridizing sequence in the two mouse strains. Eight of the 13 enzymes tested generate RFLVs; thus, the locus appears to be quite polymorphic. Because there are no consistent size differences suggestive of insertions and deletions, each of the enzymes is probably detecting point mutations. The strain distribution pattern (SDP) was determined for

IN

THE

MOUSE

311

P450IVA-EcoRI, -HpaI, and -StuI and is identical for each of these RFLVs (Table 2), indicating that the P450IVA genes are probably clustered at a single chromosomal site, which we will call Cyp4a in accord with the proposals of Nebert et al. (1991). Comparison of these SDPs with other known SDPs shows that there are O/26 recombinants with Mb-13 (Traina et aZ., 1981) and l/26 recombinants with Pmu-19 (Frankel et al., 198913). Cyp4a-Mtv-13 is 0.00-4.13 CM (P < 0.05) and Cyp4a-Pmu-19 is 0.03-6.96 CM (P < 0.05), indicating that Cyp4a is midway between brown (b) and Gpd-1 on chromosome 4 (Taylor, 1989). The Cyp4a locus has recently been mapped to chromosome 4 using a panel of mouse-hamster somatic cell hybrids (Kimura et aZ., 1989a).

Mapping

the GSTa Variants

The SDPs for the RFLVs detected by RsaI and Hind111 exhibited 5 differences among the 26 RI strains, indicating that the probe hybridizes to distinct loci in the mouse genome. The RsaI RFLV SDP differs from the dilute (d) locus in only one strain, indicating close linkage. The estimated recombination frequency is 0.010, with 95% confidence limits of 0.0003 and 0.073. The SDPs for flanking markers Xmu-15 (Frankel et al., 1989a) and Pgm-3 (Nadeau et al., 1981) suggest that the RsaI RFLV maps distal to dilute. This is consistent with the position of the Gstu locus recently mapped to the same region in an interspecific backcross (Kingsley et aZ., 1989), and very similar data reported by Kasahara et al. (1990) on BXD and other RI strains. Previously, GSTa sequences were assigned to chromosome 9 by analysis of somatic cell hybrids (Czosnek et aZ., 1984). Therefore it is reasonable to assume that the RsaI RFLV identifies the Gsta locus. Surprisingly, despite the general similarity of the RsaI and HindI SDPs, the latter does not appear to map to chromosome 9. [Kasahara et al. (1990) interpreted their data to mean that both of the BXD SDPs defined with the Gsta probe are located on chromosome 9. We are skeptical of this interpretation, because it requires postulating 4 double crossovers in the d-Pgm-3 interval. The minimum number of crossovers in the interval increases from 4 to 11 if one places both the GSTa SDPs in this region.] We provisionally designate the second SDP (our HindIII RFLV) as Gst2-rs (glutathione S-transferase a- related sequence). The closest association for Gst2-rs is with the Ode-10 locus (Richards-Smith and Elliott, 1984), with only 4 of 26 strains discordant. Ode-10 has not yet been assigned to a specific chromosome. Since the polymorphic HindI fragment is weakly hybridizing, it may represent either a previously unnoticed pseudogene of the mouse or

312

MILES

a EcoRI

BD

HpI BD

stu; BD

b

EcoRI BD

Hind111 BD

ET

AL. C

RsaI BD

KpnI % D

Hind111 B D

EcoRI % D

Hind111 BD

PstI BD

stu1 BD

KpnI % D

RsaI BD

2-

FIG. 1. Restriction Polymorphic fragments shown by filled triangles,

fragment length variants detected with (a) P450IVA, (b) GSTol, (c) GST?r, and (d) UDPGT-K39 are indicated by triangles. In the case of UDPGT-K39-Hi&II and -PstI the strongly hybridizing and the weakly hybridizing fragments by open triangles. Sizes are shown in kb.

some unknown functional gene with some degree of homology to the GSTa gene.

Mapping

the GST?r Variants

The full-length human GST?r cDNA probe detects up to 37 kb of hybridizing sequence in the C57BL/6J mouse strain and up to 35 kb in the DBA/BJ strain. Ten of the 14 enzymes tested generate RFLVs. All of the detected RFLVs appear to reflect a loss of DNA from the DBA/2J strain, suggesting that there is a deletion in this strain compared with C57BL/6J. The SDP was determined for GSTn-EcoRI, -IYindIII, -KpnI, and -RsaI, and is identical for each of these

cDNA probes. fragments are

RFLVs, possibly because each detects the same insertion/deletion variant (Table 2). Comparison of these SDPs with other known SDPs shows O/26 recombinants with Pmu-21 (Frankel et aZ., 1989b) and therefore the Gstp locus maps to the distal end of chromosome 1 (Gstp-Pmu-21 is 0.00-4.13 CM, P -c 0.05). (We propose to use the symbol Gstp to identify this locus until its relationship to the two human GSTT-encoding genes is established.)

Mapping

the UDPGT-K39

Variants

The 0.88-kb PstI-BarnHI fragment containing the 5’ half of the coding region of a cDNA encoding a phenol-metabolizing enzyme detects restriction frag-

DRUG-METABOLIZING

ENZYMES

TABLE Distribution

IN THE

313

MOUSE

2

Pattern of P46OIVA-, GSTn-, GST+, and UDPGT-K39-Detected Polymorphisms among BXD Recombinant Inbred Mice BXD

1 P450IVA Eco RI HpaI I%1 GSTa Hind111 RsaI GST?r EcoRI KpnI Hind111 RsaI UDPGT-K39 (1) Eco RI stu1 KpnI HindIII-1 PstI-1 UDPGT-K39(2) HindIII-2 PstI-2

2

5

6

8

9

11

12

13

DBDBBDBDBDDDDDBD DBDBBDBD DBDBBDBDBDDDDDBDDDBDDDDDBD

14

15

16

13

19

20

21

DDDDDB

22

23

24

25

D

DBDDDDDBD BDDDDD

27

28

29

30

31

32

D

DBBDDBDDBBDDDDDDDBBDBBDBBB DBBDDDBDBBBDDDBDDBBDBBDDBB BBDDDD BDDDBBBBDDDDBDBDBBD BBDDDDBBDDDBBBBDDDDBDBDBBD BBDDDDBBDDDBBBBDDDDBDBDBBD BBDDDDBBDDDBBBBDDDDBDBDBBD BBDDDBBBDBDBBBDBDDDDBDBDDD BBDDDBBBDB BBDDDBBBDBDBBBDBDDDDBDBDDD BBDDDBBBDBDBBBDBDDDDBDBDDD BBDDDBBBDBDBBBDBDDDDBDBDDD

BBBDBDDDDBDBDDD

BBDDDBBDBBDDBDDDDBBDBDDBBB BBDDDBBDBBDDBDDDDBBDBDDBBB

ments that hybridize strongly in both strains. (The 3’ half of the coding region for UDPGTs from different families is conserved, whereas the 5’ half is quite divergent; Tephly and Burchell, 1990). The strongly hybridizing polymorphic fragments detected with EcoRI, StuI, KpnI, HindIII, and PstI define a common SDP and presumably detect the same locus. This SDP differs by three from that of Acrg (Mock et cd., 1990) on chromosome 1 in 23 informative strains. The estimated recombination frequency between UDPGT-K39 (suggested locus symbol: Ugtlal) and Acrg is 0.041, with confidence limits of 0.0072 and 0.17. This provisionally places this locus in the proximal half of chromosome 1 between Acrg and Emu-l 7. In addition, with the enzymes NindIII and PstI, a weakly hybridizing fragment is detected in the C57BL/6J strain but is absent in DBA/M. This second variant (suggested locus symbol: Ugtlal-rs) shows l/26 recombinants with the previously mentioned O&-IO locus (Richards-Smith and Elliott, 1984). The estimated recombination frequency is 0.010, with 95% confidence limits of 0.0003 and 0.073. The Ugtlal-rs locus would appear to be on the opposite side of the Gstu-related sequence, which also belongs to this unmapped linkage group. DISCUSSION

The chromosomal locations of Cyp4u, GSTa, GSTa, and UDPGT-K39 are compared with those of

other genes encoding drug-metabolizing enzymes in Table 3. It is clear that in the mouse, as in humans, these genes are spread throughout the genome. Previously, it has been shown that a certain degree of linkage homology exists between mouse and human in the cytochrome P450 multigene family. For example, the human CYP2A and CYP2B loci are within 350 kb on chromosome 19 (Miles et aZ., 1989) and the mouse Cyp2u and Cyp2b loci are closely linked on an homologous region of chromosome 7 (Miles et al., 1990); similarly, human CYP2C maps to a region of chromosome 10 showing homology to the region of mouse chromosome 19 where Cyp2c is located (Meehan et al., 1988). In this context it is interesting to note that the human gene encoding GSTa (GST2) maps to chromosome 6~12 (Board and Webb, 1987) and that mouse chromosome 9 contains a region of synteny homology (Pgm-3 to Mod-l) close to Crbp and the Gst2 locus. Similarly, human phenol UDPGT (gene symbol GNTl) is located on human chromosome 2 (Harding et al., 1990), and there is a region of mouse chromosome 1 (I&-I to Cryg) that shows synteny homology to human chromosome 2 (Nadeau and Reiner, 1989). In humans GSTa-class genes are located on chromosomes 11 (GST3) and 12 (GST3-like) (Board et d., 1989), and the location of CYP4A is not yet known. The detection of RFLVsaand mapping of a further four loci involved in drug metabolism in the mouse will aid in the investigation of their contribution to

MILES

314 TABLE Location

3

genes to chromosome band 6~12. Proc. Natl. Acad. Sci. USA 84:2377-2381.

of Genes Encoding Drug-Metabolizing Enzymes in the Mouse

Gene/gene

cluster

PIOh.?

ET AL.

Location

3.

Means of mapping

Ref. 4.

CYPl

Mid-g, near Mpi-I

S

(53)

R

(20)

CYP20

I, near Gpi-I

S

Cyp26

7, near Gpi-1

I3 S

CypZe

19, near Got-1

R I

15

R S

(27) (35) (42) (46) (45) (33) (33) (33)

I

S

R

S Cyp2d CypZe CYM

Probably 5

S R

CYM

P450IVA

PO? Eph Gsta Gstm Gstp Ugtlol” Ugt2b”

GSTa

GSTx UDPGT-K39 UDPGT,-3

(16)

This work

6 1

S S

9

R S R I3

(46) (4‘3

near Mtv-13

1, near 1 5

Pmu-21

R R

S

!Z'oxicol.8:1-32. 5. 6.

7.

(54) (46) (46)

R S R

4,

(12) (26)

(31)

(‘3

This work

m3) This work This work (29)

Note. S, somatic cell hybrids; R, RI lines; I, In situ hybridization; B, interspecific hackcross; Par, cytochrome P450 oxidoreductase; Eph, epoxide hydratase. ’ We have adopted the nomenclature system for UDPGTs suggested by a recent international committee comprising D. R. Nelson, B. Burchell, K. W. Bock, T. Iyanagi, D. Lancet, G. J. Mulder, I. S. Owens, J. Roy Chowdhury, G. Siest, T. R. Tephly, P. I. Mackenzie, and D. W. Nebert, the proposals of which are to be published.

BOARD, P. G., WEBB, G. C., AND COGGAN, M. (1989). Isolation of a cDNA clone and localization of the human glutathione S-transferase 3 genes to chromosome bands llq13 and 12q13-14. Ann. Hum. Genet. 53: 205-213. BURCHELL, B. (1981). Identification and purification of multiple forms of UDP-glucuronosyltransferase. Reu. Biochem. BURCHELL, B., AND COUGHTRIE, M. W. H. (1989). UDP-glucuronosyltransferases. Pharmucol. Ther. 43: 261-289. CZOSNEK, H., SAFtID, S., BARKER, P. E., RUDDLE, F. H., AND DANIEL, V. (1984). Glutathione S-transferase Ya subunit is coded by a multigene family located on a single mouse chromosome. Nucleic Acids Res. 12: 4825-4833. DEJoNG, J. L., MORGENSTEF~N,R., JORNVALL, H., DEPIERRE, J. W., AND Tu, C.-P. D. (1988). Gene expression of rat and human microsomal glutathione S-transferases. J. Bid. Chem. 263:8430-8436.

DEMANT, P., AND HART, A. A. M. (1986). Recombinant congenie strains-A new tool for analyzing genetic traits determined by more than one gene. Immunogenetics 24: 416422. 9. DU~TON, G. J. (1980). “Glucuronidation of Drugs and Other Compounds,” CRC Press, Boca Raton, FL. 10. EARNSHAW, D., DALE, J. W., GOLDFARE, P. S., AND GIBSON, G. G. (1988). Differential splicing in the 3’ non-coding region of rat cytochrome P-452 (P45OIVAl) mRNA. FEBS L&t. 8.

236:357-361.

11. FEINBERG, A. P., AND VOGELSTEIN, B. (1983). A technique for radiolabeling DNA restriction endonuclease fragments to high specific activity. Anal. Biochem. 136: 6-13. 12. FRANKEL, W. N., STOYE, J. P., TAYLOR, B. A., AND COFFIN, J. M. (1989a). Genetic analysis of endogenous xenotropic murine leukemia viruses: Association with two common mouse mutations and the viral restriction locus Fv-1. J. Virol. 63: 1763-1774. 13. FRAN-L, W. N., STOYE, J. P., TAYLOR, B. A., AND COFFIN, J. M. (1989b). Genetic analysis of endogenous polytropic proviruses by using recombinant inbred mice. J. Virul. 63: 38103821.

14.

chemical carcinogenesis ena.

and other complex phenom-

ACKNOWLEDGMENTS We thank Dr. G. Bulfield for supplying some of the inbred mice and genomic DNA. We are grateful to those people who supplied DNA probes: Dr. G. G. Gibson, P450IVA, Professor M. Muramatsu, GSTa; Dr. C. Kasper, P450 reductase; Dr. F. J. Gonzalez, P450IID; and Drs. J. Taylor and S. Pemble, microsomal GST and GSTp. This work was supported in part by NIH Grants GM18684 and CA33093. REFERENCES 1. AYESH, R. IDLE, J. R., RITCHIE, J. C., CROTHERS, M. J., AND HE-L, M. R. (1984). Metabolic oxidation phenotypes as markers for susceptibility to lung cancer. Nature 312: 169170. 2. BOARD, P. G., AND WEBB, G. C. (1987). Isolation of a cDNA clone and localization of human glutathione S-transferase 2

FRANKLIN, R. A., AND PARKE, D. V. (Eds.) (1986). Human genetic variation in oxidative drug metabolism. Xenobiotica 16:367-509.

15.

GONZALEZ, F. J. (1989). The molecular biology of cytochrome P45Os. Pharmacol. Rev. 40: 243-266. 16. GONZALEZ, F. J., MATSUNAGA, T., NAGATA, K., MEYER, U. A., NEBERT, D. W., PASTEWSKA, J., KOZAK, C. A., GILLE?TE, J. R., GELBOIN, H. V., AND HARDWICK, J. P. (1987). Debrisoquine 4-hydroxylase: Characterization of a new P450 gene subfamily, regulation, chromosomal mapping, and molecular analysis of the DA rat polymorphism. DNA 8: 149161. 17. GONZALEZ, F. J., SKODA, R. C., K~URA, S., UMENO, M., ZANGER, U. M., NEBERT, D. W., GELBOIN, H. V., HAF~~~ICK, J. P., AND hlEyER, U. A. (1988). Characterization of the common defect in humans deficient in debrisoquine metabolism. Nature 331: 442-446. 18. GOUGH, A. C., MILES, J. S., SPURR, N. K., Moss, J. E., GAEDIGK, A., EICHELBAUM, M., AND WOLF, C. R. (1990). Identification of the primary gene defect at the cytochrome P450 CYPSD locus. Nature 347: 773-776. 19. HARDING, D., JEREML~H, S. J., POVEY, S., AND BURCHELL, B. (1990). Chromosomal mapping of a human phenol UDP-

DRUG-METABOLIZING

20.

21.

22.

23.

24.

25.

26.

27.

28.

29.

30.

31. 32.

glucuronoeyl transferaae, GNTl. Ann. Hum. &met. 64: 1721. HILDEBRAND, C. E., GONZALEZ, F. J., KOZAK, C. A., AND NEBERT, D. W. (1985). Regional linkage of the dioxin-inducible P-450 gene family on mouse chromosome 9. B&hem. Biophys. Res. Commun. 130: 396-406. HILL, R. E., SHAW, P. H., BARTH, R. K., AND HASTIE, N. D. (1985). A genetic locus closely linked to a proteaae inhibitor gene complex controls the level of multiple RNA species. Mol. Cell. Biol. 6: 2114-2122. IYANAGI, T., WATANABE, T., AND UCHNAMA, V. (1989). The 3-methylcholanthrene-inducible UDP-glucuronosyltransferase deficiency in the hyperbilirubinemic rat (Gunn rat) is caused by a -1 frameshift mutation. J. Biol. Chem. 254: 21302-21307. JACKSON, M. R., AND BURCHELL, B. (1986). The full length coding sequence of rat liver androsterone UDP-glucuronyltransferase cDNA and comparison with other members of this gene family. Nucleic Acids Res. 14: 779-795. KANO, T., SAKAI, M., AND MURAMATSU, M. (1987). Structure and expression of a human z class glutathione S-transferase messenger RNA. Cancer Res. 41: 5626-5630. KASAHARA, M. E., MATSUMARA, E., WEBB, G., BOARD, P. G., FIGUEROA, F., AND KLEIN, J. (1990). Mapping of class alpha glutathione S-transferase 2 (Gst-2) genes to the vicinity of the d locus on mouee chromosome 9. Gerwmics 8: 90-96. KIMURA, S., HARDWICK, J. P., KOZAK, C. A., AND GONZALEZ, F. J. (1989a). The rat clofibrate-inducible CYPIA subfamily. II. cDNA sequence of IVAB, mapping of the Cy& locus to mouse chromosome 4, and coordinate and tissue-specific regulation of the CYP4A genes. DNA 8: 517-525. KIMURA, S., KOZAK, C. A., AND GONZALEZ, F. J. (1989b). Identification of a novel cytochrome P450 expressed in rat lung: cDNA cloning and sequence, chromosome mapping and induction by 3-methylcholanthrene. Biochemistry 28: 37983803. KINGSLEY, D. M., JENKINS, N. A., AND COPELAND, N. G. (1989). A molecular genetic linkage map of mouse chromosome 9 with regional localizations for Gstu, T3g, Ets-I and Ldlr loci. Genetics 123: 165-172. KRASNEWICH, D., KOZAK, C. A., NEBERT, D. W., AND MACKENZIE, P. J. (1987). Localization of UDP glucuronosyltransferase gene(s) on mouse chromosome 5. Somat. Cell Mol. Genet. 13: 179-182. LEWIS, A. D., HICKSON, I. D., ROBSON, C. N., HARRIS, A. L., HAYES, J. D., GRIFFITHS, S. A., MANSON, M. M., HALL, A. E., MOSS, J. E., AND WOLF, C. R. (1988). Amplification and increased expression of alpha class glutathione S-transferaseencoding genes associated with resistance to nitrogen mustards. Proc. Natl. Acad. Sci. USA 85: 8511-8515. LYMAN, S. D., POLAND, A., AND TAYLOR, B. A. (1980). Genetic polymorphism of microsomal epoxide hydrolase activity in the mouse. J. Biol. Chem. 255: 8650-8654. MANNERVIK, B., AND DANIELSON, U. H. (1988). Glutathione transferases-Structure and catalytic activity. CRC Crit. Reu. Biochem.

33.

Acad.

35.

Sci.

USA

MssHAN, R., AND WOLF, C. R. (1989). Close linkage of the human cytochrome P450IIA and P450IIB gene subfamilies: Implications for the assignment of substrate specificities. Nucleic Acids Res. 17: 2907-2917. MILES, J. S., Moss, J. E., w, R. R., AND WOLF, C. R. (1990). Close linkage of the cytochrome P45OIIA gene subfamily (Cyp2a) to Cyp26 and Coh on mouse chromosome 7. Genomics

7: 445-448.

MOCK, B., KRALL, M., BLACKWELL, J., O’BRIEN, A., SCHURR, E., GROS, P., SKAMENE, E., AND Pcrrrsn, M. (1990). A genetic map of mouse chromosome 1 near the Lsh-Zty-Bca disease resistance locus. Genomics 7: 57-64. 37. NADEAU, J. H., KOMPF, J., SIEBERT, G., AND TAYJ.,OR,B. A. (1981). Linkage of Pgm-3 in the house mouse and homologies of three phosphoglucomutase loci in mouse and man. Bio&em. Genet. 19: 465-474. 38. NADEAU, J. H., AND REINER, A. H. (1989). Linkage and synteny homologies in mouee and man. In “Genetic Variants and Strains of the Laboratory Mouse” (M. F. Lyon and A. G. Searle, Eda.) 2nd ed., pp. 506-536, Oxford Univ. Press, London/New York. 39. NEBERT, D. W., NELSON, D. R., COON, M. J., ESTABROOK, R. W., FEYEREISEN, R., FUJII-KURNAMA, Y., GONZALEZ, F. J., GUENGERICH, F. P., GUNSALUS, I. C., JOHNSON, E. F., LOPER, J. C., SATO, R., WATERMAN, M. R., ANJJWAXMAN, D. J. (1991). The P450 superfamily: Update on new sequences, gene mapping and recommended nomenclature. DNA 10: 1-14. 40. PORTER, T. D., ANLI RASPER, C. B. (1985). Coding nucleotide sequence of rat NADPH-cytochrome P-450 oxidoreductase cDNA and identification of flavin-binding domains. Proc. Natl.

41.

42.

43.

44.

45.

46.

47.

48.

49.

50.

Acad.

Sci.

USA

82: 973-977.

RICHARDS-SMITH, B., AND ELLIOT, R. (1984). Mapping of a family of repeated sequences in the mouse genome. Mouse News Z&t. 71: 4647. SAUNDERS, A. M., AND SELDIN, M. F. (1990). A molecular genetic linkage map of mouse chromosome 7. Genomics 8: 525-535. SEIDEGARD, J., PERO, R. W., MILLER, D. G., AND BEAM, E. J. (1986). A glutathione transferase in human leukocytes as a marker for the susceptibility to lung cancer. Carcinogencsis 7: 751-753. SILVER, J. (1985). Confidence limits for estimates of gene linkage based on analysis of recombinant inbred strains. J. Hered.

86: 2662-2666.

MILES, J. S., BICKMORE, W., BROOK, J. D., MCLAREN, A. W.,

315

IN THE MOUSE

36.

23: 283-337.

MEEHAN, R. R., SPEED, R. M., GOSDEN, J. R., ROUT, D., HUTTON, J. J., TAYLOR, B. A., HILKENS, J., KROEZEN, V., HILGERS, J., ADESNIK, M., FRIEDBERG, T., HASTIE, N. D., AND WOLF, C. R. (1988). Chromosomal organization of the cytochrome P450-2C gene family in the mouse: A locus associated with constitutive aryl hydrocarbon hydroxylase. Proc. Natl.

34.

ENZYMES

76: 436-440.

SIMMONS, D. L., AND KASPER, C. B. (1983). Genetic polymorphisms for a phenobarbital-inducible cytochrome P-450 map to the Coh locus in mice. J. Biol. Chem. 258: 9585-9588. SIMMONS, D. L., LALLEY, P. A., AND KASPER, C. B. (1985). Chromosomal assignments of genes coding for components of the mixed-function oxidase system in mice. J. Biol. Chem. 260: 515-521. SOUTHERN, E. M. (1975). Detection of specific sequences among DNA fragments separated by gel electrophoresis. J. Mol. Biol. 98: 503-518. TAYLOR, B. A. (1978). Recombinant inbred strains: Use in gene mapping. In “Origins of Inbred Mice” (H. C. Morse, Ed.), pp. 423-428, Academic Press, New York. TAYLOR, B. A. (1989). Recombinant inbred strains. In “Genetic Variants and Strains of the Laboratory Mouse” (M. F. Lyon and A. G. Searle, Ede.), 2nd ed., pp. 773-796, Oxford Univ. Press, London/New York. TAYLOR, J. B., OLIVER, J., SWINGTON, R., AND PEMBLE,

316

MILES

S. E. (1991). Structure of human glutathione S-transferase mu genes. Biochem. J. 274: 587-593. 51. TEPHLY, T. R., AND BURCHELL, B. (1990). UDP-glucuronosyltransferases: A family of detoxifying enzymes. Trends Pharmacol. Sci. 11: 216-279. 52. TRAINA, V. L., TAYLOR, B. A., AND COHEN, J. C. (1981). Genetic mapping of endogenous mouse mammary tumour viruses: Locus characterization, segregation and chromosomal distribution. J. Viral. 40: 735-744. 53.

TUKEY, R. H., LALLEY, P. A., AND NEBERT, D. W. (1984).

ET AL. Localization of cytochrome Pi-450 and Pa-450 genes to mouse chromosome 9. Proc. N&l. Acad. Sci. USA 81: 31633166. 54. UMENO, M., SONG, B.-J., KOZAK, C. A., GELBOIN, H. V., AND GONZALEZ, F. J. (1988). The rat P450IIEl gene: Complete intron and exon sequence, chromosome mapping, and correlation of developmental expression with specific 5’ cytosine demethylation. J. Biol. Chem. 263: 4956-4962. 55. WOLF, C. R. (1986). Cytochrome P-4509: Polymorphic multigene families involved in carcinogen activation. Trends Genet. 2: 209-214.