Mammalian Genome 8, Brief Data Reports

A.

Pll �9 [] [] �9 []

A g t r l a � 9 [] �9 [][]

Dl3Mit4 �9 [] �9 [] �9

N u m b e r o f M i c e 43 47 2 1 l

B~

D3MiI60 �9 [] [] []

Agtrlb �9 [] �9 []

o, , ir~ �9 [] �9 �9

Number of Mice 43 49 1 l

Fig 1. (A) Localization of Agtrla to proximal Chr 13. The markers Mf4 (mesoderm/mesenchyme fork head 4) and D13Bir8 had not recombined with Agtrla. They were, therefore, placed at the same location as Agtrla, and are not shown. (B) Localization of Agtrlb to proximal Chr 3. The filled boxes indicate the presence of both C57BL/6J and SPRET/Ei alleles. The open boxes indicate the presence of only SPRET/Ei alleles. The number of offspring carrying each type of chromosome is indicated at the bottom. Complete raw data for the Jackson BSS backcross are available on The World Wide Web at the URL address http://www.jax.org/resources/ documents/cmdata.

A T l b are encoded by separate genes, Agtrla and Agtrlb respec- tively.

In this study, we used probes specific to the mouse Agtrla and Ag#lb genes to follow allelic segregation in an interspecific back- cross panel Comparison of the haplotype distribution pattern lo- cated Agtrla to proximal Chr 13, and Agtrlb to proximal Chr 3. Map Manager [8] was used to determine linkage, to calculate marker distances, and to draw haplotypes. The most likely marker order was determined by inspection of the data and minimization of double recombinants (Fig. 1). Three recombinants were found between Agtrla and the placental lactogen-1 gene (PI1), and one recombinant was found between Agtrla and D13Mit4. These data place Agtrla 3.2 cM distal to Pll and 1.1 cM proximal to D13Mit4 on Chr 13. One recombinant was identified between Agtrlb and D3Mit60 and between Agtrlb and D3Bir3 respectively. Thus, Agtrlb was placed 1.1 cM distal to D3Mit60, and 1.1 cM proximal to D3Bir3 on Chr 3.

The human AGTR1 gene has been mapped to Chr 3q21-25. The mouse Agtrlb gene maps to proximal Chr 3. Proximal Chr 3 contains other genes that map to human 3q21-25, such as carboxy- peptidase A3 (Cpa3) [9,10]. This study, therefore, extends the relationship between these chromosomes.

Acknowledgments: We thank Lucy Rowe for her advice and for critical reading of the manuscript. We also thank Achim Gossler for manuscript review and Sylvia Hiller for her assistance.

References 1. Rowe LB, Nadeau JH, Turner R, Frankel WN, Letts VA, Eppig JT, Ko

MSH, Thurston SJ, Birkenmeier EH (1994) Mamm Genome 5, 253- 274

2. Murphy TJ, Alexander RW, Griendting KK, Runge SR, Bemstein KE (1991) Nature 35l, 233-236

3. Gemmill RM, Drabkin HA (1991) Cytogenet Cell Genet 57, 162-166 4. Szpirer C, Riviere M, Szpirer J, Levan G, Guo DF, Iwai N, Inagami T

(1993) J Hypertens 11,919-925 5. Hall J (1986) Am J Physiol 250, R960-R972 6. Elton TS, Stephan CC, Taylor GR, Kimball MG, Martin MM, Durand

JN, Oparil S (1992) Biochem Biophys Res Commun 184, 1067-1073 7. Sasamura H, Hein L, Krieger JE, Pratt RE, Kobilka BK, Dzau VJ

(1992) Biochem Biophys Res Commun 185, 253-259 8. Manly KF (1993) Mamm Genome 4, 303-313 9. Reynolds DS, Gurley DS, Austen KF (1992) J Clin Invest 89, 273-282

295

10. Gurish MF, Nadeau JH, Johnson KR, McNeil HP, Grattan KM, Austen KF, Stevens RL (1993) J Biol Chem 268, 11372-11379

An M M T V integration site maps near the distal end of mouse Chromosome 11

Lakshmi Rajah, Jaquelin P. Dudley

Department of Microbiology, University of Texas at Austin, Austin, Texas 78712, USA

Received: 13 October 1996 / Accepted: 9 November 1996

Species: Mouse Locus name: A somatically acquired MMTV integration site from a T-cell lymphoma Locus symbol: Pad3 Map position: Pad3 is located on mouse Chromosome (Chr) 11: Centromere-(//)-DllHun20-(1.09 + 1.08)-D11Hun21-(2.13 + 1.49)-Fasn-Hfh4-Pad3-D11Xrf16-DllXrf107 (upper 95% confidence limit is 3.8 cM). Method of mapping: Southern blot analysis of DNAs from 94 backcross (N2) progeny from the cross (C57BL/6JEi x SPRET/Ei) x SPRET/Ei (The Jackson laboratory BSS panel). Database deposit information: MGD accession number MGD- CREX-706 Molecular reagents: The Pad3 probe is a 0.7-kb Pstl unique cel- lular DNA fragment 3' to an acquired MMTV provirus from the P, Lc~ 1 T-cell lymphoma. Allele detection: By Southem blot analysis, the Pad3 probe hy- bridized to an EcoRl fragment of approximately 10 kb in C57BL/ 6J (Mus musculus) DNA and 5 kb in SPRET/Ei (Mus spretus) DNA. Previously identified homologs: None. Discussion: Mouse mammary tumor virus (MMTV)" is a type B retrovirus that induces breast cancer and, at a lower frequency, T-cell tumors in mice [1]. MMTV-induced breast cancers result from the transcriptional activation of cellular oncogenes located in the vicinity of proviral integration sites. Activation of transcription at nine different loci--namely, Intl/Wntl, Int2/Fgf3, Int3/Notch- like, lnt4/Wnt3, F gf-4/Hst/K-FGF/Int5/lntH/aromatase, Fgf8, Int6, and WntlOb---has been observed in MMTV-induced mammary tumors in mice [24] . However, the mechanism of T-cell tumor induction by MMTV is still unclear. TBLV, a T-cell tropic infec- tious variant of MMTV, has been shown to integrate on the mouse X Chr and to activate transcription of one or more nearby genes [5].

To determine if the Pad3 MMTV integration site maps close to any known cellular gene, we used RFLPs for the Pad3 locus to screen the BSS panel [6]. Digestion of each of the 94 BSS panel DNAs with EcoRI detected a 5-kb band in the homozygous ani- mals and both 5- and 10-kb bands in the heterozygous animals. Thus, the allelic segregation of the Pad3 locus in the BSS panel was determined. This pattem was compared with the patterns of segregation of the approximately 2000 other loci previously mapped in the cross, and linkage was found to distal Chr 11 [6]. The Pad3 locus cosegregated with at least four other markers, namely, Fasn, Hfh4, DllXrfl6, and DllXrfl07 (see Fig. 1). The proximity of the Pad3 integration to Hfh4 (HNF-3/forkhead ho- molog 4) is intriguing, since members of the HNF-3/forkhead or "winged hel ix" proteins are transcription factors with diverse bio- logical functions [7-9]. At least two members of the forkhead family have been shown to participate in translocation-mediated oncogenesis in human malignancies. Translocations to FKHR (also designated ALV) have been observed in alveolar rhabdomyo-

Correspondence to: J. Dudley

296 Mammalian Genome 8, Brief Data Reports

Jackson BSS

'.Brcal, Grn, Pea3 .D11Hun18

-D11Bit14, D11MitlO I

3 cMI -D11Bit15

-D11Hun20 -D 11Hun21

.Fasn, Hfh4, Pad3, D11Xrf16, D11Xrfl07

Fig. 1. Localization of Pad3 on the distal end of mouse Chr 11 with The Jackson Laboratory BSS panel [7]. The map is depicted with the centro- mere toward the top. A 3-cM scale bar is shown to the left of the figure. It appears that Pad3 is among the cluster of markers that is the most distal on the Jackson BSS Chr 11 map.

sarcomas [10], and a chimeric fusion protein involving the AFXI gene has been implicated in an acute lymphocytic leukemia [11]. In addition, the avian retroviral oncogene qin is a member of the forkhead gene family [11]. The possibility that the Hfh4 gene may be affected by MMTV integration at the Pad3 locus is being tested.

References 1. Dudley J, Risser R (1984) J Virol 49, 92-101 2. Marchetti A, Buttitta E, Miyazaki S, Gallahan D, Smith GH, Callahan

R (1995) J Virol 69, 1932-1938 3. MacArthur CA, Shankar DB, Shackleford GM (1995) J Virol 69,

2501-2507 4. Lee FS, Lane TF, Kuo A, Shackleford GM, Leder P (1995) Proc Natl

Acad Sci USA 92, 2268-2272 5. Mueller RE, Baggio L, Kozak CA, Ball JK (1992) Virology 191,

628~537 6. Rowe LB, Nadeau JH, Turner R, Frankel WN, Letts VA, Eppig JT, Ko

MSH, Thurston SJ, Birkenmeier EH (1994) Mamm Genome 5, 253- 274

7. Mouse Genome Database (MGD), Mouse Genome Informatics, The Jackson Laboratory, Bar Harbor, Maine. World Wide Web (URL: http://www.informatics.jax.org/). (4 September 1996)

8. Lai E, Clark KL, Burley SK, Darnell JE Jr. (1993) Proc Natl Acad Sci USA 90, 10421-10423

9. Avraham KB, Fletcher C, Overdier DG, Clevidence DE, Lai E, Costa RH, Jenkins NA, Copeland NG (1995) Genomics 25, 388-393

10. Fredericks WJ, Galili N, Mukhopadhyay S, Rovera G, Bennicelli J, Barr FG, Rauscher FJ III (1995) Mol Cell Biol 15, 1522-1535

11. Parry P, Wei Y, Evans G (1994) Genes Chromosomes Cancer 11, 79-84

A second locus encoding elevated phosphoglycerate mutase activity (Pgam2e) maps to mouse Chromosome 4

Walter Pretsch, Jack Favor

GSF-National Research Center for Environment and Health, Institute of Mammalian Genetics, Ingolst~idter Landstral3e 1, D-85764 Neuherberg, Germany

Received: 24 September 1996 / Accepted: 1 November 1996

Species: Mus musculus Locus name: Phosphoglycerate mutase 2 elevated Locus symbol: Pgam2e Map position: Centromere-D4MiH2-(6.1 + 2.4)-D4Mit203-(3.1 + 1 .7) -D4Mit204- (4 .1 + 2 .0) -D4Mit134- (1 .O + 1 . 0 ) -

Correspondence to." W. Pretsch

6.1

3.1

4.1

1.0 1.0

4.1

3.1

D4Mitl2

D4Mit203

D4Mit204

D4Mit134 Pgam2e

D4Mit54

D4Mit158 D4Mitl70

D4Mit189

Fig. 1. A partial map of mouse Chr 4 with the distances in cM between each pair of loci.

Pgam2elNeu-(1.O + l.O)-D4Mit54-(4.1 + 2.0)-D4Mit158- D4Mit170-(3.1 + 1.7)-D4Mit189 (distances in cM) (Fig. 1). Method of mapping: Pgam2e 1Ne" was localized by haplotype analysis of 98 progeny from an intraspecific backcross, (B6C3F 1- Pgam2elUe"/+) x C57BL/6 employing eight Chromosome (Chr) 4 microsatellite markers (Fig. 2). Allele detection: The specific phosphoglycerate mutase (PGAM; EC 2.7.5.3) activity was measured in erythrocytes [1], and off- spring were categorized for their mutation genotype. Previously identified homologs: Human PGAMA has been as- signed to 10q25.3 [2]. Discussion: Recently, we described the ethylnitrosourea(ENU)- induced mutation PGAM12601 causing a 300--400% PGAM ac- tivity elevation in blood compared with wild-type animals; segre- gation analysis localized the corresponding gene Pgamle to the medial region of Chr 19 [3]. Peters and Ball described an ENU- induced mutation, pge, with 25 to 30-fold higher erythrocyte PGAM activity [4]. Homozygotes are fully viable and fertile, This regulatory locus is not linked to ru or ep on Chr 19, indicating that there exists at least one additional locus for elevated PGAM ac- tivity.

A mouse mutant (PGAM46) with elevated PGAM activity was recovered in the offspring of an irradiated male mouse [5]. From the present linkage data, we designate the mutation Pgam2e 1Ne". The mutation causes PGAM activity elevation to 244 + 27% (n = 30) in blood of heterozygotes as compared with wild-type animals at weaning. The increased PGAM activity has not been observed in liver, lung, kidneys, spleen, heart, or brain of heterozygotes; the enzyme shows no significant heat stability alteration in blood at 64~ Other than the enzyme activity effects, there were no de- tectable visible phenotypes associated with Pgam2e in heterozy- gotes. Homozygous mutants are lethal. Thus, mutants Pgamle 1Ne"

D4Mitl2 D4Mit203 D4Mit204 D4Mit134 pgam2e D4_Mit54 D4Mit158 D4Mitl70 D4Mit189

IB --,IBIBBI B IB IBI 'IBIIB

, , i B 36 40 2 4 2 1 2 2 1 1 4 3

Fig. 2. Haplotype data for the intraspecific backcross showing the markers flanking the Pgam2e locus segregating on Chr 4. The number of backcross progeny for each haplotype are identified by the number beneath the col- umns. Alleles are designated by the black (C3H) and white (C57BL/6) squares.