GENOMICS

11,

709-719

(1991)

A Map of 22 Loci on Human Chromosome

22

JAN P. DUMANSKI,*+ ELISABETH CARLBOM,t’$ V. PETER COLLINS,* MAGNUS NORDENSKJ&D,t BEVERLY 5. EMANUEL,~ MARCIA L. BUDARF,~~ HEATHER E. MCDERMID,~T ROGER WOLFF,** PETER O’CONNELL,** RAY WHITE,** JEAN-MARK LALOUEL,** AND MARK LEPPERT** *Ludwig Institute for Cancer Research, Stockholm Branch, t Department of Clinical Genetics, and *Department of Neurosurgery, Karolinska Hospital, S-104 07 Stockholm, Sweden; §Department of Human Genetics and “Department of Pediatrics, University of Pennsylvania School of Medicine, and Division of Human Genetics and Molecular Biology at the Children’s Hospital of Philadelphia, Philadelphia, Pennsylvania; lIDepartment of Human Genetics, University of Alberta, Edmonton, Alberta, Canada T6G2E9; and **Howard Hughes Medical Institute and Department of Human Genetics, University of Utah, Salt Lake City, Utah 84 132 Received

November

29, 1990;

July 2, 1991

number of disease-susceptibility genes, e.g., genes involved in diGeorge syndrome, Cat-Eye syndrome, and recessive congenital methemoglobinemia (RCM). It is also the site of a number of constitutional translocations and of (11;22) translocations associated with Ewing sarcoma (McKusick, 1987). A large number of other expressed genes have also been assigned to this chromosome (Kaplan and Emanuel, 1989). Two chromosome 22 genes involved in oncogenesis have been cloned and deleterious mutations characterized at the molecular level; namely, the breakpoint cluster region (bcr) gene involved in the (9;22) translocations in chronic myeloid leukemia cells and the immunoglobulin X-chain locus involved in the variant (8;22) translocations in Burkitt lymphoma (Kaplan et aZ., 1987). Among the disease-susceptibility genes located on this chromosome is the unknown gene involved in the origin of central neurofibromatosis (NF2), a condition inherited as an autosomal dominant trait. This gene has been placed between two DNA markers through linkage studies in a single large affected family (Rouleau et al., 1990). A loss of chromosome 22 alleles has been observed in sporadic acoustic neurinomas (Seizinger et al., 1986) as well as in one acoustic neurinoma, one neurofibroma, and one meningioma from subjects affected with NF2 (Seizinger et al., 1987). On the basis of these findings it has been proposed that a common pathogenetic mechanism, affecting a locus on chromosome 22, is involved in NF2 and meningioma (Seizinger et at., 1987). However, recent findings based on the deletion mapping of chromosome 22 in meningiomas suggest that the meningioma and NF2 loci are separate entities (Dumanski et al., 1990b). A chromosome 22 linkage map with a large number of ordered loci within the regions of interest for NF2 and meningioma should greatly facilitate further investigations.

We constructed a genetic linkage map of the entire long arm of human chromosome 22 with 30 polymorphic markers, defining 22 loci. The map consists of a continuous linkage group 110 CM long, when male and female recombination fractions are combined; average distance between the loci is 6.2 CM. All loci were placed on the map with high support against alternative orders (odds in excess of 1OOO:l). The order of loci presented in our map is in full agreement with that of the previous linkage maps of chromosome 22 and with the physical assignment of markers. Two markers included in this map, III-831 (0225222) and pEFZ31 (D22S32), allowed us to better define the region of the (11;22) translocation breakpoint specific for Ewing sarcoma. Ten additional polymorphic markers were placed on the 22-loci map with odds lower than 1OOO:l against alternative locations. In total, we have introduced 29 new markers on the linkage map of chromosome 22. 0 lvel Academic

revised

Press, Inc.

INTRODUCTION

In growing numbers, cloned DNA segments that detect DNA sequence polymorphisms are forming a powerful set of tools for studies in human genetics. Genetic linkage maps of DNA markers have already been constructed for most of the human chromosomes. Such maps allow one to locate a disease gene between flanking markers, as the first step toward cloning of the gene and characterization of an underlying genetic defect. A large number of disease-susceptibility genes have been precisely localized using this strategy, and several have been cloned and characterized at the molecular level. Chromosome 22 is the second smallest of the human autosomes and contains approximately 52 Mb of DNA (Kaplan et al., 1987). It harbors a substantial 709

All

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

710

DUMANSKI

Three published linkage studies of human chromosome 22 (Donis-Keller et al., 1987; Julier et aZ., 1988; Rouleau et aZ., 1989) give consistent results with respect to gene order. However, a number of gaps remained not covered by markers included in these previous maps, and the markers used in those studies were typed only on a part of the set of CEPH (Centre d’Etude du Polymorphisme Humain, Paris) families or on a set of families that was not a part of CEPH reference panel. A large set of new chromosome 22-specific polymorphic DNA markers has been developed over the past few years (Budarf et al., 1991; Dumanski et al., 1990a). In the present linkage study of the long arm of human chromosome 22 we have included many of these new probes in addition to probes used for previously published linkage maps. We have been able to refine the localization of the (11;22) translocation breakpoint that is specific for Ewing sarcoma. Furthermore, markers ordered in the linkage map presented here are likely to contribute to the better localization of the gene at the origin of central neurofibromatosis through linkage analysis of affected families. This map will also allow refined deletion-mapping studies of tumor-associated loss of chromosome 22 alleles in meningiomas and acoustic neurinomas. The large set of markers along the entire long arm of chromosome 22 should allow detection of linkage for any disease-susceptibility gene that resides on this chromosome. Finally, the map provides anchor points for a 1-CM high-resolution linkage map of chromosome 22. MATERIAL

DNA Markers

AND

METHODS

and Determination

of Genotypes

A total of 40 polymorphic, chromosome 22-specific DNA markers were used in the present study (see Table 1 for description of marker/enzyme systems). Anonymous markers W13E (D22S21), WllOD (D22922), W24F (D22823), and W22D (D22S29) were kindly provided by G. Rouleau. They were developed from a flow-sorted chromosome 22-specific DNA library and mapped to chromosome 22 by genetic linkage (Rouleau et al., 1989). The probe for platelet-derived growth factor p chain (PDGFB), pSM-1 (Fourney et al., 1988; Ratner et al., 1985), was obtained from ATCC. The probes for the myoglobin gene (MB), pHM27.B2.9 (Julier et aZ., 1988; Weller et aZ., 1984); the arylsulfatase gene (ARSA), pCP8 (Herzog et aZ., 1990); the immunoglobulin X polypeptide, variable region (IGLV), pV3.3 (Julier et al., 1988); the BCR gene (5’ bcr, 1.95-kb HindIII-BglII fragment) (Roschmann et al., 1987); the cytochrome P450 gene, subfamily IID, debrisoquine/sparteine type (CYP2D), dbl (Gonzalez et al., 1988; Skoda et al., 1988); and the anonymous markers p22/34 (D22S9,

ET

AL.

McDermid et aZ., 1986) and FZIXCll (D22S171) were kindly provided by A. Jeffreys and N. Blin, J.-C. Kaplan, G. Assum, F. Gonzalez, B. N. White, and G. Thomas, respectively. The anonymous markers pMS3-18 (D22S1, Barker et al., 1984) and pEFZ31 (D22S32, Krapcho et al., 1988) were published previously. Twenty-one other anonymous markers, which are designated by KI numbers, were developed from the DNA library mentioned above and were mapped to three different regions of the long arm of chromosome 22 on a panel of somatic cell hybrids: KI-169 (D22S209), KI-780 (D22S213), KI-831 (D22S212), and KI-1582 (D22S208) to region 22qll-q12; KI-185 (D22S90), KI-844 (D22Sl59), KI-261 (D22Sl58), KI474 (D22993), KI-117 (D22986), KI-711 (D22S160), KI-106 (D22885), KI-436 (D229102), KI-211 (D22S91), KI-120 (D22S87), and KI-218 (D22S92) to region 22q12-q13; KI-839 (D22S95), KI-260 (D22997), KI-216 (D22S84), KI-63 (D22382), KI-536 (D228157), and KI-1105 (D22S94) to region 22q13qter (Dumanski et aE., 1990a; Carlbom et al., 1991). Four anonymous markers, pH4la (D22345), pH19 (022840) pH91 (D22955), and pH130 (D22864), were developed from the same library and assigned to chromosome 22 on a panel of somatic cell hybrids within the region distal to the Ewing sarcoma (11;22) translocation breakpoint (Budarf et al., 1991). Marker RWClO (D22S219) was isolated from a total human cosmid library using bovine /3 crystalline cDNA probe llC8 (Hogg et aZ., 1987), which was a generous gift from M. Gorin. Genotypes were collected from a reference panel of 60 three-generation pedigrees. Forty of these families are part of the CEPH reference panel. Extraction of DNA from lymphoblastoid cell lines was performed according to the method of Bell et al. (1981). Restriction endonuclease digestion, Southern transfer of the DNA; preparation of phage, plasmid, and cosmid DNA; and radiolabeling of the probes with 32P followed methods described elsewhere (Barker et al., 1984; Cavenee et al., 1984; Dumanski, 1990; Feinberg and Vogelstein, 1984; Reed and Mann, 1985). Hybridizations were carried out using whole vector and insert constructs, or insert DNA only, in low-meltingpoint (LMP) agarose, except for marker KI-63 (022382). A 0.55-kb fragment from the 2.1-kb insert of this marker was used for hybridization experiments, because it detects only a single constant band in addition to variable bands (see Table 1). After PCR amplification of the insert (Dumanski, 1990), TaqI digestion of the reaction product, and electrophoresis through LMP agarose, a 0.55-kb band was excised from the gel and used for radiolabeling (Feinberg and Vogelstein, 1984). Genotypic data were entered into a computer database. These entries were checked against the original autoradiograms to verify geno-

A MAP

type interpretations entry.

OF

22 LOCI

and the absence of errors

ON

in data

Linkage Analysis Linkage analysis was performed with the LINKAGE computer program package (Lathrop et al, 1985). A genetic map of chromosome 22 was constructed following an iterative algorithm previously described (White et al., 1990). Briefly, six loci were selected after inspection of all pairwise recombination estimates automatically arrayed so as to best display clustering. These loci were selected on the basis of their high heterozygozity and their apparent regularity of spacing. This initial order was validated by testing all possible permutations. Additional loci were progressively added to the map by iterative application of the following scheme: (1) a maximum location score on the current map is derived for each unmapped locus; any locus with a likely location in an interval with odds in excess of 1OOO:l against alternative intervals becomes a candidate for addition to the map; (2) only one locus can be added in every other interval of the map at each cycle; (3) in any interval, the locus added to the map presents odds in excess of 1OOO:l when alternative orders are tested and these odds are greater than those obtained for other candidates for this interval; (4) map intervals are reestimated, adjacent loci are retested, and another cycle is performed unless no addition to the map can be made under the conditions imposed. We calculated the odds against the inversion of adjacent loci using up to three flanking loci (three on each side of the inverted pair) (Lathrop et al., 1984, 1986). Evidence for sex-specific differences in recombination rates was investigated under two models: no sex-difference (combined male and female recombination rates) and sex-difference variable from one interval to another. RESULTS

Characterization

of the Polymorphic

Loci

Table 1 summarizes the polymorphic marker/enzyme systems used for construction of the new linkage map of chromosome 22. We typed, on average (for all polymorphic marker/enzyme systems), 712 individuals of a maximum 844 from the panel of reference pedigrees. The majority of the probes detect two-allele polymorphic marker/enzyme systems each of which reflects a single change in a restriction endonuclease recognition site. Two probes, pH4la (022845) and pH91 (D22855), detect VNTRs with five and four alleles, respectively. Six markers (IGLV, BCR, PDGFB, CYP2D, RWClO (D22S219), and ARSA) represent gene loci, and marker KI-436 (D22SlO2) detects evolutionarily conserved DNA sequences.

HUMAN

CHROMOSOME

22

711

Three markers-W13E (D22S21), W24F (D22S23), and W22D (D22S29)-that detect multiple marker/ enzyme systems were haplotyped prior to the linkage analysis. The observed heterozygosities ranged from 0.2 to 0.58 with an average of 0.45 (see Table 1).

Linkage Results On the basis of results from pairwise linkage analysis of all markers, six loci (BCR, W22D (D22S29), PDGFB, KI-260 (0229971, W13E (D22S21), and ARSA) were chosen as anchor points for the map. After all different permutations of these markers were tested, they were ordered with high odds against the next most probable order (102?1). Other markers were added to the map following an iterative algorithm described under Materials and Methods. Markers that were in complete linkage (0% recombination with high lod scores) were constantly placed within the same interval of flanking loci, and displayed consistent recombination events with flanking loci were combined into haplotypes: W22D (D22S29), KI-106 (DZZSSS), KI-117 (D22S86), and KI-711 (D22S160); KI-185 (D22S90) and KI-211 (D22S91); PDGFB and KI-218 (022992); KI-839 (D22S95) and pH130 (022864); and WllOD (D22S22) and KI-1105 (022394) see Tables 1 and 2, and Fig. 1). Two of these haplotypes involved the initial anchor points (W22D and PDGF-B) and confirmed the order of our trial map with even higher odds (1031:1 and greater). The iterative mapping procedure was pursued until no other locus could be added to the map with the tolerance specified. This analysis yielded a linear order for 22 different loci on the long arm of human chromosome 22; 30 different polymorphic markers were used for the construction of this map (see Fig. 1). For all loci included in the map, odds against alternative locations were in excess of 1OOO:l. The overall support of this map against alternative orders is high (odds greater than 1.3 X 104:1), as determined by inversion tests of adjacent loci on the map (see Fig. 1). The linkage group spans 110 CM for the combined-sex model of recombination frequencies (see Table 3), with an average distance between the loci of 5.2 CM. Ten additional markers typed on the panel of reference pedigrees could not be placed on the established map of 22 loci showed in Fig. 1 within a single interval, with odds >lOOO:l against alternative locations. Figure 2 indicates the intervals of their most likely locations. Two anonymous markers included in the present study, KI-63 (022882) and FZIXCll (D22Sl71), displayed complete linkage with a maximum lod score of 144. The genotypes displayed with Ta.qI polymorphisms identified by both markers, as well as the position of allelic bands on the same set of Southern filters, were identical for more than 800 individuals

712

DUMANSKI

TABLE Description Locus

Probeb locus

I.D.” 1 A

2 3 B 4 C D 5 6 E 7.1

7.2 7.3 7.4 7HAP 8 9 F 10 11.1 11.2 1lHAP 12.1 12.2 12HAP G 13.1 13.2

of the Polymorphic, Insert size (kb)

p22/34 (30) D22S9 pv3.3 (22) IGLV 5’ BCR (36) HindIII-BgZII KI-169’ (6a) D22S209 RWClO’ D22S219 KI-1582’ (6a) D22S208 pMS3-18 (2) D22Sl KI-780’ (6a) D22S213 KI-831’ (6a) D22S212 pEFZ31 (25)’ D22S32 KI-844 (13) D22S159 W22D (37) D22S29

pSM-1 PDGFB KI-218 D22S92

KI-120 (13) D22S87 KI-839 (13)” D22S95 pH130 (4) D22S64

1.85

TaqI

-

3.3

KpnI

1.95

TaqI

3.9

TagI

40

MspI

1.7

B&II

0.58

BglII

1.8

TaqI

2.9

PVUII

3.1

MspI

1.4

XbaI

10.0

TaqI

4.8

ToqI

5.5

MspI

3.8

MspI

Haplotype-Markers 3.8 KpnI (44,22)

(34, 17) (13)

Constant bands (kb)

7.0

Td

4.0

MspI

2.1

TagI

6.2

TaqI

6.7

MspI

Haplotype-Markers 2.6 Hind111 1.1

MspI

Haplotype-Markers 0.73

MspI

2.0

EcoRV

2.8

PvuII

AL.

1

22-Specific

Enzyme

fr.

KI-106 (13)’ D22S85 KI-117 (13)’ D22S86 KI-711 (13)’ D22S160 KI-261 (13) D22S158 pHM27.B2.9 MB KI-474 (13) D22S93 KI-436 (13)” D22S102 KI-185 (13) D22S90 KI-211 (13) D22S91

Chromosome

ET

DNA

Markers

Allele sizes (kb)

5.8 3.2 19 17.5 14.1 5.2 1.95 1.2, 0.75 2.1, 1.5, 0.9: 3.8 3.2 0.7 Many 3.5 3.0 4.5 4.3 9.5 6.5 1.8, 0.68 3.5 2.7 3.3, 0.6 2.3 1.8 3.5 2.6 16 15 4.0, 2.6 Al 5.1 A2 4.3, 0.8 Bl 6.4 B2 2.6 Haplotype 2.5 4.9 4.0, 0.9 1.5 9.5 6.8 8.5, 1.8 4.0 2.8 W22D, KI-106, KI-117, and KI-711 18 7.5 3.7, 2.1 1.6 1.3 2.5, 1.0 3.3 2.45 5.0 4.1 3.8 7.7 6.3, 1.4 2.5, 1.4, 1.3, 1.95 0.9, 0.7 1.1 KI-185 and KI-211 21 14.5, 6.5 0.75 0.7 0.6 PDGFB and KI-218 4.5 4.2 20 15 1.8 1.6 1.5

Used Allele frequenciesd 0.66 0.34 0.26 0.46 0.28 0.68 0.32 0.49 0.51 0.33 0.67 0.54 0.46 0.65 0.35 0.36 0.64 0.43 0.57 0.63 0.37 0.41 0.59 0.42 0.58 0.04 0.96

(0.03) (0.03) (0.03) (0.03) (0.03) (0.03) (0.03) (0.04) (0.04) (0.04) (0.04) (0.04) (0.04) (0.05) (0.05) (0.03) (0.03) (0.04) (0.04) (0.03) (0.03) (0.03) (0.03) (0.03) (0.03) (0.01) (0.01)

0.27 0.73 0.7 0.3 0.11 0.89

(0.03) (0.03) (0.03) (0.03) (0.02) (0.02)

0.52 0.48 0.34 0.66 0.76 0.24 0.49 0.51 0.52 0.48 0.14 0.86

(0.03) (0.03) (0.03) (0.03) (0.03) (0.03) (0.04) (0.04) (0.03) (0.03) (0.02) (0.02)

0.74 0.26 0.53 0.47

(0.03) (0.03) (0.03) (0.03)

0.89 0.11 0.47 0.53 0.36 0.64

(0.02) (0.02) (0.03) (0.03) (0.03) (0.03)

in the Present Observed heterozygozity

Study Number typed’

0.46

802 (316)

0.72

504 (269)

0.51

778 (386)

0.46

713 (278)

0.49

504 (192)

0.45

473 (185)

0.49

267 (98)

0.44

753 (335)

0.52

657 (296)

0.45

718 (260)

0.57

609 (293)

0.55

733 (363)

0.07

790 (68)

0.58 0.44

(396) 746 (335)

0.48

740 (296)

0.18

812 (157)

0.92 0.57

789 (383)

0.45

821 (351)

0.38

809 (327)

0.57

688 (312)

0.45

812 (358)

0.26

640 (187)

0.58 0.4

755 (294)

0.49

774 (353)

0.67 0.22

765 (120)

0.49

736 (328)

0.48

781 (337)

A MAP

OF

22 LOCI

ON

TABLE Locus I.D.” 13HAP H

14 15 16.1 16.2 16HAP I 17.1 17.2 18 19 J

Probeb locus

p45Odbl CYPBD

(18,41)

pHl9 (4) D22S40 KI-260 (13)” D22S97 KI-1105 (13)’ D22S94 WllOD (37) D22S22 KI-536 (13) D22Sl57 KI-63 (13) D22S82 FZIXCll (9) D22S171 KI-216 (13) D22S84 pH4la (4) D22S45 W24F (37) D22S23

Insert size (kb)

Enzyme

Haplotype-Markers 1.568 EcoRV

5.0

Hind111

4.1

TaqI

1.95

-11

1.9

Td

Hanlotwe-Markers 22 -EcoRV 0.55

TWI

0.53

TaqI

3.0

PVUII

1.4

PVUII

2.1

Td EcoRV

20

21 22

W13E (37) D22S21

pH91 (4) D22S55 pCP8 (20) ARSA

3.6

Td

5.5

TasI

1.4

TaqI

HUMAN

CHROMOSOME

l-Continued

Constant bands (kb)

Allele sizes (kb)

KI-839 14.5

and pH130 44 29 12 11.7 6.2 2.3 2.6 2.4 6.6 5.5 2.2 1.9 1.8 KI-1105 and WllOD 6.5 5.4, 1.1 3.2 1.2 1.6 3.0 1.8 3.9 2.2, 1.7 1.8 VNTR, five alleles 2.7-2.0 3.6 2.1, 1.5 6.8 5.4 Haplotype Al 2.8 1.05 A2 1.6 Bl 1.5 B2 1.35 Haplotype 2.2, 0.56 VNTR, four alleles 1.55-1.25 1.1 3.9 3.2

’ Polymorphic markers are identified by numbers or letters and are in agreement with those nomenclature is according to HGMlO. b Parenthetical numbers in this column indicate references for the markers used. ’ Indicates markers containing repetitive DNA elements. d Parenthetical numbers in this column denote + standard deviation. ’ Parenthetical numbers in this column denote the number of potentially informative meiosis gous parent.

scored (see Tables 1 and 2 and Fig. 1, locus numbers 17.1 and 17.2; autoradiograms not shown). These two chromosome 22-specific probes, isolated independently in two laboratories, therefore identify the same TuqI polymorphism and represent the same locus.

Sex-Specijic Differences in Recombination Frequencies No significant differences were observed in the overall lengths of maps constructed on the basis of sex-specific recombination fractions (107 CM in male, 116 CM in female, and 110 CM for the combined male

713

22

Allele frequenciesd

0.08 0.9 0.02 0.54 0.46 0.44 0.56 0.79 0.21 0.31 0.69

(0.02) (0.02) (0.01) (0.04) (0.04) (0.03) (0.03) (0.03) (0.03) (0.03) (0.03)

0.17 0.83 0.42 0.58 0.42 0.58 0.41 0.59

(0.02) (0.02) (0.03) (0.03) (0.03) (0.03) (0.03) (0.03)

0.1 0.9 0.76 0.24

(0.02) (0.02) (0.04) (0.04)

0.8 0.2 0.86 0.14

(0.03) (0.03) (0.02) (0.02)

Observed heterozygozity

0.48 (0.03) 0.52 (0.03) in the figures

from

pedigrees

0.74 0.20

540 (102)

0.48

408 (143)

0.57

816 (423)

0.36

806 (305)

0.39

685 (241)

0.62 0.25

792 (215)

0.55

808 (394)

0.56

815 (402)

0.43

737 (314)

0.49

808 (372)

0.14

812 (135)

0.26

447 (100)

0.38 0.35

804 (290)

0.28

826 (251)

0.57 0.34

777 (266)

0.47

753 (327)

and in Tables

with

Number typed’

at least

(261)

(406)

2 and 3. The

one heterozy-

and female map). Inspection of these maps did suggest an excess of male over female recombination within three regions of the chromosome (see Table 3). Of these, one is at the telomere of the long arm of chromosome 22 and is composed of the four contiguous intervals between marker KI-216 (D22S84), locus 18 and the ARSA gene, locus 22. However, a clear excess of female over male recombination was observed in two other regions, between loci 5 and 6 and 9 and 10 (see Table 3). We tested whether sex differences in recombination frequency were statistically significant within each interval of the map. After allowance for multiple tests, significant values were ob-

6 2 4 1 1 1 1 1 0 1 0 I 0 1 29 I 5 0

I

0 1 I

1 2 0 1 0 0 1

16.2 I 17.1

17.2

19 I 20 2, 22

,a

10

2 3 B 4 c D 5 6 E 7HAP 7.1 7.2 1.3 7.4 8 9 F 10 llW1 (I.1 11.2 12HAP1 12.1 12.2 G 13~1 13.1 13.2 H 14 15 ,b"mO 16.1

0 0 1 1 0

0 2 2

I 0

17

*

I

A

1 I I 0 I 0 0

1 I I

7 9 3 7 ‘I 4 3 4 4 7 6 3 5 2 4 1 2 1

25 23 14 9 3 ,I 8 7 4 22 10 7 6 2

.

A

LJXUSI

I,

5 12 14 7 20 1 23 I, 7 9 3 12 5 11 5 7 4

16

10

2 1 5 . 10 3 5 7 4 7 13 4 7 3 2 3 1 3 0 7 2 4 4 2 4 3 1 0 1 0 3 3 0 1 0 0 1 1 0 1 1 I 0 0

17

B

8 5 3 . 4 11 IO 2 4 10 3 6 3 2 5 3 6 2 5 3 2 4 2 3 1 4 3 3 0 2 1 I 2 0 0 0 0 0 0 0 0 0 0

28 II

4

5 10 0 . 13 2 9 6 10 6 4 3 0 10 2 7 4 a 3 5 1 1 3 1 3 1 2 I 0 1 1 2 0 0 0 0 0 2 0 0 0 0

,a 1,

3,

c

17

2, 23 25 5 24 17 13

7

22 13 9 15 9 13 1 8 3 6

18 23 2.5 8 21 10 I,

6

1, II 9 11 9 14 7 14 9

22

5

4

7 6 3 3 2 1 3 3 3 2 3 1

2 5

6

3 2 1 1 1 1 1 2 0 2 0 0

3 0 1 1 1 I 1 2 1 1 0 0

9

1

3

6

4

8

4 1, 8

7

2, 5 32 I2 10 9 20 IO 14 11 9

26

16 46

2) 51

6 * 23 48

1, 7 4 3 5 5 2 5 3 5 0 0

9

19

8

20 I1 13 18 8 12 7 23 11 I2

19

30 34 33 IO 38 3, 2,

9 4 * b4

19 14 13 9 4

14

8 lb 23 9 I

4, 26

E

40 22

6

19

38 19 20 15 17 12 22 4

5

30

2,.

40 17 ,2 12 9 10 3 *

D

2, 14 a 9 14 14 9 12 4 13 4 3

3,

22

8

44 39

6,

52 32 38 23

36

72 50

38

117 92 107 4, a3 73 69

7 5 2 .

17 17 18 12 6

,a

40 19

3,

2 3 4 4 4 3 7 1 4

i

6

1 1

7 6

a

4

17 I2

29

7 2 3 6 6 5 7 1

12 10

10

4

15 ,a

13

2,

16

15

24 14

10

29 24

14

0 * 48 5 40 24 22

7 4 1 0

21 I5

2,

40 22

23

* 36 40 1, 38 3, 26

8 6 2 0

I5

14 5

10 7

38 2, 20 19 16

7.2

1,

16 19

,a

4, 19

'IHAP 7.1

Recombination

1 I

5

-

3 3 3 10 IO 4 4 I

16 6

13

6

23 15

28

IO

16 14

23

14

3, 19

I2

0 0 I5 39 33 23

4 3 I 0

17 5

16

43 22 2, 17 22

7.3

5 2 2 4 4 5 4 1 3 2 1

6

6

0 6

I, 5

14

16

12 3 5 9 9 6 10 0 9 5 0

7 9 16 12

16

1,

32

29 18 I5 I2

12 13 7 a 3

a

34 2,

16

18

2 3 * 30 2,

0 a 19 I2 1

1

1

I I

7

7

2

7

35 20 17 16 21 14

8

0

0

2 0

9

6

14

67

35 25 18 1, 11 15

7.4

9 4 5 10 10 4 7 2 8 3 I

4 16 13 10

19

15

2,

29 14 17 ,

18

34 18

15

3 1 5 * 3,

4

2

3 3

8

13

12

16

37 23 2, 22 68 22

9

Estimates

9 6 5 9 9 5 5 0 5 0 0

6 I, 6 13

1,

10

2,

29 14 18 9

8

29 23

13

2 0 0 0 -

0

1

1, 1, I 1

13

7

26 19 17 16 24 11

F

6 a 9 9 5 9 5 9 3 2

7 9 ,5 I5 10

20

23

42

38 1, 25 12

I5

40 26

-

11 13 9 a 10

8

6

I5 16 5 9

I5

14

43 28 23 25 59 14

10

among

38

15

4

4

6

6 4 3 9 9 10 13 5 12

15 I2

5

6

28

46 1, 56 26

59 28

4

lb

6

is 15 13 22

29 23 lb 7 9

12

77 4, 56 25 70 37 37

56

* 110

0 .

9

7 1,,22,14543

8

27 28 25 22 I, 15 I2 13 9 8 8 7 6 a 9

29

6

5 ,

6

2, 25 20 2, 10 I, 1, I, 8 7 7 8

26

39

lIH.,P 11.1

0

1

4 4 b 8 I 5

6

9 23 17 10 3 5 ,3 12 I, 3

1,

0 0 . 28'0 15

4

13

35

1,

3

10

18 18 1, 14 8 13

7

* 3, 1, 42 22 22 8 1, 20 25 10 14

1 2 2

10 10

2

2

9 9 10 I, 5 ,

9

2 1 4 0 0 f 24 64 35 3, 14 12 1, 18 8 8

9 10

9 10 10 9 ,I

I,

9 11 10 7 9

10

13

,4

18

22 27 24 22 17

29

13

14

,8

16

28 27 30 25 29

24

48

12.2 42

1

2

8 8 5 6 4 8

1

I 1 0 0 0 0 * 29 22 II ,I 10 12 I, 9 5

a

10

2, 27 20 22 12 23

1,

3 2 3 2 2 1 0 * 114 105 25 22 49 40 20 24

10 13

1, 10

9 12 1, 10

18 23 12

17

24 27 35 26 25

29

5 8 0 10 9 6

Used

3

5

12

6

19 19 14 12

10

4 3 4 2 3 1 0 0 * 39 17 15 21 23 II 12

1, I5

10 10 9 13

I, 10

17 27 12

2,

27 37 35 25 28

32

43

,3HAP 13.1

6 6

9 76 3

I2

17 26 20 18 18

13

26

G

22 Loci

6

6

1,

6

14 14 7 9

7

3 2 4 2 1 2 0 0 0 * 9 I5 ?.a 20 IO 13

9 9

10 13 15 10

1, 8

20 20 11

15

18 20 3, 23 22

25

43

13.2

2

0

1

2

5 5 3 3

0

10 0

3

12 5 4 I, 1, 4 12

,a 14

9 7 5 19 .

6 a 4 10 4

13 10 15

11

35 24 24 23 24 22 68 17 22 23 3 13 13 13 8 I2 12 8

14

likelihood

11 10 II 4 3 4 0 0 0 0 * I 3 7 1 3

I, 12

10 1, 74 9

14 14

I1 26 3

22

18 25 48 30 18

23

24

H

42 23 23 13 43 43 2, 30 I8 29 2, 9

lb IO

9

6

9 I, a 10

,I

31 30 25 25 2, 18 lb 17 2, 14 18 15 18 22

26

5, 40 32 35

15

in the Present

the diagonal are the maximum to each recombination fraction.

24 24 19 20 10 19

16

9, 116 33 93 54 45 18 2, 30 42 24 22

2 2 2

10 11

13 13 12 13 12 10 10

5 2 2 5 2 0 5 1 5

16

19

17

8

6

2x5 28 30 24 26

32

39

,ZHAP 12.1

9

18 24 12 18 12

22

4,

II1

the Chromosome

2

Note. Locus identification numbers or letters correspond to those in Table 1. Numbers above multiplied by 100. The numbers under the diagonal are the maximum lod score corresponding

1 6 2 4 1 2 3 0 0 0 0 0 0 0 I 0 0

6

2

5 2 5 2

6

9 4 7 5 13 7 9 3 5 3 4 4 I I 3 3 2 2 0 2 0 I

14 7 12 II 24 10 8 9 4 14 6 LO 7 12 5

a

29 8 4 *

3

23 5 35

2

Pairwise

TABLE

10 5 0 * 25 36 25 25 24 3, 1, 30 27 7

estimates

6,

18

9 ,I 10 18 10 12 12 12

12 I, 10 14 1, 1, 12 1, 12 5 95 8A 4, 59 59 38 45 28 4, 32 17

19

16

22 28 ,9 66 ,a 2, 24 2, 19 19 14 ,a 19 16 14

16

30

32

37 30 36 30 23 27 17 22 20 24 24 20 18 20 20 14 14

38 37 27

36

46 34 29

16"AP 16.1

Study

20 20 1, 1, 18 20 20 5

6 1 0

I5 2, 12 13 15 29 13

14

19 13 12

16 14

58 35 lb 11 23 22 23 2, 24 20 23 2, 18

36

72

36 36 60

39

I

144 35 45 17 37 1, 8

4 2 2

13 12 13 19 13 4

1, I8 11

20 ,a 14

13 18

,a 2, 2, 22

,b I5 16

13 14 13 14 19 18 8 4 4 4 3 4 4 * 34 I5 38 17 14

I, 13 12 13 19 ,3 4 I 4 2 2 0 * 35 45 1, 3, 1, 8

22 2, 23 19 18 1, 19

ii 2, 22 13 ii 20 18 14 ,I 18

17

27 25

19

34 29 34 30 25

39

37 44 64 35

35

3,

18

,I(

29 2, 2,

26

26

38 33 34 25 24

46

34 43 34 47

38

42

17.2

29

38 33 34 25 24

46

34 43 34 47

38

42

17.1

of the recombination

18 35 35 19 1.5 13 18 13 6

12 12 10 I2 9 19 14 5 0 0

10

10

16

1,

11 16

2, 23 23 2n

28

24 23 2.4

,a

24 28

58 65 47 50 34

5, 33 32

16.2

27 23 19

4 4

3, 3, 34 65 32 55 2, 23 23 26 20 14 22 I5 20 20 20 19 23 11 8 12 9

36

33

2,

35

26

55

44 43

55 55 34

46

60

J

fractions,

20 18 18 20 23 I5 a 7 a 1 5 3 3 3 * 30 45 w 2,

20 16 2,

19 2, 29 2, 19 20 20

24

25 29

25

35 29 32 25 28

29

43 35 43

4, 36

57

19

20 20 2, 32 14 12 8 8 10

16

2, 18 25

24

25 24 30 19 22 2,

26

37 42 35 45 43 34 35 40 27 28 28 30 28

39

45

20

2,

1, -

60 43 52 47 41 46 4" 42 29 42 39 33 34 37 36 29 27 31 59 24 28 29 29 23 19 32 23 22 25 20 50 70 a 7 6 10 2 10 10 9 9 6 5

39 48 40 35 37 42 36 34 38 34 39 26 3, 30 43 30 27 32 29 28 31 21 25 31 18 lb 23 19 20 17 17 12 9 8 16 17

64 56 41 54 42 40 45 38

22

A MAP

OF

22 LOCI

ON

tained for the two regions with an excess of female over male recombination (between loci 5 and 6, xf = 11.8; between loci 9 and 10, xt = 13.3, P < 0.005) and in one of the regions with apparent excess of male recombination (between loci 20 and 21, xf = 8.92, P

< 0.005). Refinement of the Localization of Ewing Sarcoma (11;22) Translocation Breakpoint A translocation breakpoint (11;22) specific to Ewing sarcoma was previously localized on the long arm of chromosome 22 between flanking markers pMS318 (D22S1) on the centromeric side and markers pEFZ31(022832), DP22 (D22S15), and leukemia inhibitory factor gene (LIF) on the telomeric side (Budarf et al., 1989a,b; Zhang et aZ., 1990a,b). Of these markers, pMS3-18 (D22SI) and pEFZ31 (022S32) were included in the present study. We have been able to introduce at least one new locus, KI-831 (022S212), to the region between markers pMS3-18 (D22SI) and pEFZ31(022832). This marker was previously mapped on a panel of somatic cell hybrids to a region on the centromeric side of the Ewing sarcoma (11;22) translocation breakpoint (Dumanski et aZ., 1990a). Consequently, the breakpoint is located between the proximal marker KI-831 (022S212) and the distal marker pEFZ31 (022S32), which bracket a region of 4 CM (see Figs. 1 and 2). DISCUSSION Three previously published linkage maps of chromosome 22 (Donis-Keller et al., 1987; Julier et al., 1988; Rouleau et al., 1989) were constructed from 4,5, and 13 polymorphic loci, respectively. Results with regard to the gene order of loci common to these studies were consistent. For the linkage study presented here we used a total of 40 polymorphic markers. The order of 22 loci presented in Fig. 1 and the 10 additional loci showed in Fig. 2 is in full agreement with the previous, less detailed reports. We have significantly reduced the gaps that existed between the breakpoint cluster region gene (BCR) and marker W13E (022321). The largest gap between any two markers over the continuous region of 66 CM between BCR and W13E (022S21) is now 10 CM, with an average of 3.6 CM. Over certain parts of the chromosome, e.g., the 32-CM segment between loci KI-436 (0229102) and pH91 (022855), our map is of even higher density; the average distance between loci is 2.9 CM in this region. The most telomeric marker reported in previous chromosome 22 linkage studies was W13E (D22S21). Our map introduces two new loci, ARSA and pH91 (D22S55), distal to W13E (D22S21), that extend the map by 18 CM toward the telomere of the long arm of chromosome 22. The aryl-

HUMAN

CHROMOSOME

22

715

sulfatase A gene (ARSA), the most telomeric marker, has been physically assigned to the region 22q13.31qter (Kaplan et al., 1987). The most centromeric marker reported in this study is p22/34 (D22S9), which was physically assigned to the vicinity of the centromere of chromosome 22 (McDermid et aZ., 1986). Thus, our linkage map spans the entire long arm of chromosome 22; no markers specific for the short arm of this chromosome are included. The largest gap on our map is the 26 CM between p22/34 (D22S9) and BCR in the centromeric part of the long arm of chromosome 22 because relatively few polymorphic DNA markers specific for this region of the chromosome were available for our study. The 110 CM covered by the total linkage group presented here (all meioses combined) is the largest yet published for chromosome 22. As mentioned above, we have extended the linkage map of the long arm of this chromosome by 18 CM toward the telomere. However, our estimate of the genetic length of 22q is not larger than that of Rouleau et al. (1989), if one only compares the loci used in both studies. In total, we have introduced 29 new markers on the chromosome 22 linkage map. Of all the linkage maps for human chromosomes published to date, it appears that the map of chromosome 22 now has the highest density of markers over a continuous region. We have found statistically significant differences in the recombination frequency between the sexes in two regions of the long arm of chromosome 22 (see Results). This is a new finding for human chromosome 22, as no such differences were previously reported (Donis-Keller et aZ., 1987; Julier et al., 1988; Rouleau et al., 1989). The excess of male over female recombination has been found to occur close to the telomere and the excess of female over male recombination occurred in the central part of the long arm of this chromosome. A similar distribution of excess of male over female recombination in the telomeric regions with a converse relationship in the central section has been reported for several other human chromosomes, such as chromosomes 11,12, and 17 (White et al., 1985; O’Connell et al., 1987; Nakamura et al., 1988). The 21 polymorphic DNA markers designated by KI numbers had been assigned to chromosome 22 on a panel of five somatic cell hybrids, and sublocalized to four regions of this chromosome (Dumanski et aZ., 1990a). Breakpoints that define the borders of the regions are within the 22qll band (the (9;22) Philadelphia translocation breakpoint), the 22q12 band (the (11;22) Ewing sarcoma translocation breakpoint), and the 22q13 band (a constitutional (1;22) translocation breakpoint). In the present study, we applied markers assigned to three of the four regions (22qll12, 22q12-13, and 22ql3-qter; see Material and Methods), as only the probes localized to these re-

716

DUMANSKI

ET

AL.

8 ~22134

(1)

.20

/ BCR (2) 11.2

KI-169

11.1 11.1

.OS

(3)

KI-lS82

.OS

(4)

.09 KI-831

11.2

12.1 12.2 12.3

(5)

MB

.04 .04 .Ol .03

(9)

.05 KI-185

(10)

KI-436

-i

4: .o 1 .04 .04

KI-839 PHl30

13.2

.04 .o 1 ::: .04 .04

.12

lo301 I 1 -2

I -3 I

S.9x104:I I -4

1

I

I

107:1

108:1 I

-5

2.2r104:1 1

I

-6

-7

I

-8

I

IO241

lo241 1

-9

I

-10

h

-1i

-12-13

I

1o12:1

I

1

lo’“:l

loll:1

1013:1

13-1;--1;-*k--l;-,=-2b--2; I

I

I

107:1

loll:1

1.3x104:1

lOlO. I

I

I

IO361

1071

I

-22 I

I

1o19:1

1

I

lo’%

I

I

lo241

1

1

102?l

FIG. 1. Combined (no sex difference in recombination frequencies) genetic map of the long arm of human chromosome 22. The map is scaled in Morgans according to the Haldane’s mapping function and spans in total 110 CM. Recombination fraction (g), which is derived from multipoint recombination estimates, is indicated for each interval (see Table 3). The original recombination fractions between the six anchor points of the map-loci 2,7,12,15,20, and 22-were 0.130,0.114,0.092,0.125, and 0.149, respectively. Known physical localisations of loci p22/34 (D22S9), PDGFB, and ARSA are indicated on the karyogram. Locus numbers in parentheses reflect numbers shown in Tables 1,2, and 3. Below: Odds against inversion of adjacent loci are indicated on the brackets.

gions order with from ability

were tested for their detection of RFLPs. The of loci presented in our linkage map agrees fully the physical assignments of markers derived the panel of cell hybrids and confirms the reliof the cell panel (Dumanski et aZ., 1990a).

The remarkable regularity with which the reciprocal translocation (11;22)(q24;q12) is found in Ewing sarcoma tumor cells (Aurias et aZ., 1983; Heim and Mitelman, 1987; Turc-Care1 et al., 1983) suggests that this rearrangement plays an important role in onco-

A MAP

TABLE

OF

22 LOCI

ON

3

Pairwise Recombination Estimates for Each Interval of the Map Displayed in Fig. 1 under Different Models of Sex-Specific Recombination

Locus number

No sex difference (combined recombination frequencies) 9

1 2 3 4 5 6 7 8 9 10 11 12 13 14 15 16 17 18 19 20 21 22

0.203 0.052 0.045 0.089 0.040 0.044 0.010 0.028 0.051 0.019 0.018 0.012 0.044 0.044 0.038 0.012 0.025 0.022 0.039 0.036 0.121

Variable sex difference in each interval Male 0

Female 0

0.168 0.046 0.039 0.099 0.000 0.051 0.016’ 0.013 0.009 0.000 0.010 0.016" 0.044 0.040 0.043 0.005 0.012 0.031" 0.039” 0.084" 0.185'

0.237 0.057 0.053 0.086 o.05gb 0.047 0.005 0.037 0.089 o.04zb 0.026 0.008 0.039 0.052 0.033 0.018 0.023 0.013 0.036 0.004 0.069

HUMAN

CHROMOSOME

717

22

large, affected family (Rouleau et al., 1990). A closer approach to this gene will require placement of a number of probes on the chromosome 22 linkage map between markers pMS3-18 (D22Sl) and W23C (D22S28)/W22D (D22S29), to allow better positioning of the NF2 gene and to facilitate presymptomatic diagnosis of NF2. Our linkage map introduces at least two new loci (pEFZ31 and KI-831) within this region of chromosome 22 (see Figs. 1 and 2). Meningioma is a common, usually benign tumor of the meninges covering the central nervous system. Meningioma cells often show monosomy 22, or a loss of a distal part of the long arm of one chromosome 22 (Zankl and Zang, 1980). This tumor is frequent in patients with the central (NF2) form of neurofibromatosis (Eldridge, 1981; Lott and Richardson, 1981).

o=d t

IGLV 42O:l

IA)

2 D Intervals * Intervals

with with

an excess of male over female recombination. clear excess of female over male recombination.

3 lo?1 8:l pMS3-,

8 (C)

I

4 ..

. . .. . . .

I

genesis. A rearrangement, apparently the same, is seen in other types of tumors, i.e., Askin’s tumor (De Chadarevian et al., 1984), esthesioneuroblastoma (Whang-Peng et al., 1987), and neuroendocrine tumor of the small intestine (Vigfusson et al., 1986). In this study we have been able to better define the position of that breakpoint between two flanking markers KI831 (DZZS’212) and pEFZ31 (022S32), which delineate a ~-CM region. This localization may be precise enough to permit long-range physical mapping of the breakpoint and molecular cloning from this region of the chromosome. The form of neurofibromatosis affecting predominantly the central nervous system (NF2) is a relatively rare, autosomal dominant disease whose hallmark is bilateral acoustic neurinoma (Eldridge, 1981). Sporadic acoustic neurinomas, as well as acoustic neurinomas from NF2 patients, often display a specific loss of chromosome 22 alleles (Seizinger et al., 1986, 1987). The gene responsible for NF2 has been localized to a relatively large region (13 CM) of chromosome 22 between markers pMS3-18 (D22S1) and W23C (D22S28) through linkage analysis of a single,

2OO:l . . .. ..I... . . . .. .. ..I.................

5

6

.. . ..KI-780 3:1 1.2x106:1 .. . . .. . . .

. RWC,Q

(B)

2.6xlO’:l

(D)

. . . . . . .. KI-844

(E)

7 KI-474

(F)

1

. . . . .. . . .. .

3x108:1

lO’!l (-yqD l.5x104:1

(H)

t’: . .. . . . . .is. . I 14

. . . ...KI- 120 (G) 3OOO:l t

1s 16 i: 19 20

I

. .. ..KI-53$(1) 2.4110 W24F

:1

(J). 700:1

21

22 FIG. 2. Regional localization of 10 additional markers on the map of 22 loci from Fig. 1. Bars on the side of the map indicate the interval(s) of most probable location. When more than one interval is shown, a dotted line indicates the most probable localization. Locus numbers to the left of the map and letters (in parentheses) agree with designations in Fig. 1 and Tables 1, 2, and 3.

718

DUMANSKI

However, cases of familial meningioma in patients without symptoms of neurofibromatosis are very rare and it is unlikely that linkage studies of meningioma families will contribute to the precise localization of the meningioma gene. Studies of allelic deletions from chromosome 22 in DNA from the tumor tissue are an alternative approach. An important prerequisite for such studies is that a sufficient number of DNA markers be ordered along the chromosome. The putative meningioma gene has been localized by deletion mapping in tumors to the region distal to the myoglobin (MB) locus on the long arm of chromosome 22, which corresponds to 22q12.3-qter (Dumanski et al., 1987,199Ob). Further studies are clearly necessary to better delimit the region containing the gene. The same strategy may be used for deletion mapping of the chromosome 22 locus involved in the origin of NF2/ acoustic neurinoma. The present linkage map introduces 17 new polymorphic loci in the region distal to MB, which is of interest for meningioma, (see Figs. 1 and 2) as well as two new loci in the NF2 region (see above). These new markers should facilitate the identification of small deletions in DNA from meningiomas and acoustic neurinomas; such deletions may pinpoint the exact location of the geneswhose mutant alleles involved in the oncogenesis of each of these tumors.

ET AL.

5.

6.

6a.

7.

8.

9.

10.

ACKNOWLEDGMENTS We are grateful to T. Elsner, M. Mitchell, L. Nelson, R. Olsen, B. Otterud, and D. Stauffer for excellent technical and computational assistance and to R. Foltz for editorial help in preparation of this manuscript. We also thank Drs. G. Assum, N. Blin, A. Jeffreys, F. Gonzalez, J.-C. Kaplan, G. Rouleau, G. Thomas, and B. N. White, as well as the American Type Culture Collection, for providing some of the recombinant DNA probes used in the present study. This work was supported in part by grants from the Swedish Cancer Society, Axe1 and Margaret Ax:son Johnson’s Fund, Erik and Edit Fernstrom Foundation, The King Gustav V’s Jubilee Fond, and Swedish Medical Research Council. R.W. and J.-M.L. are investigators at the Howard Hughes Medical Institute. Note added in proof. The polymorphic DNA markers designated by KI numbers have been submitted to American Type Culture Collection.

REFERENCES 1. AURIAS, A., RIMBAUT, C., BUFFE, D., DUBOUSSET, D., AND MAZABRAUD, A. (1983). Chromosomal translocations in Ewing’s sarcoma. N. Engl. J. Med. 309: 496-497. 2. BARKER, D., SCHAFER, M., AND WHITE, R. (1984). Restriction sites containing CpG show a higher frequency of polymorphism in human DNA. CeU 36: 131-138. 3. BELL, G., KARAM, J., AND R-R, W. (1981). Polymorphic DNA region adjacent to the 5’ end of the human insulin gene. Proc.

Natl.

Acad.

Sci. USA

78: 5759-5763.

4. BUDARF, M. L., MCDERMID, H. E., SELLINGER, B., ANKI E-L, B. S. (1991). Isolation and regional localization of

11.

35 unique anonymous DNA markers for human chromosome 22. Gerwmics 10: 996-1002. BUDARF, M., EMANUEL, B. S., MOHANDAS, T., GOEDDEL, D. V., AND LOWE, D. G. (1989a). Human differentiation stimulating factor (leukemia inhibitory factor, human interleukin DA) gene maps distal to the Ewing sarcoma breakpoint on 22q. Cytogenet. Cell Genet. 52: 1922. BUDARF, M., SELLINGER, B., GRIFFIN, C., AND EMANUEL, B. (1989b). Comparative mapping of the constitutional and tumor-associated 11;22 translocations. Am. J. Hum. Genet. 45: 128-139. CARLBOM, E., SUGAWA, N., LARSSON, C., SCAMBLER, P. J., DUMANSKI, J. P., COLLINS, V. P., AND NORDENSKJBLD, M. (1991). Identification of twelve new RFLP-markers on chromosome 22qll-qter. Hum. Genet., in press. CAVENEE, W., LEACH, R., MOHANDM, T., PEARSON, P., AND WHITE, R. (1984). Isolation and regional localization of DNA segments revealing polymorphic loci from human chromosome 13. Am. J. Hum. Genet. 36: 10-24. DE CHADAREVIAN, J. P., VEKEMANS, M., ANIJ SEEMAYER, T. A. (1984). Reciprocal translocation in small-cell sarcomas. N. Engl. J. Med. 311: 1702-1703. DELATTRE, O., AZAMBUJA, C. J., AURIAS, A., ZUCKMAN, J., PETER, M., ZHANG, F., HORS-CAYLA, M. C., ROULEAU, G. A., AND THOMAS, G. (1991). Mapping of human chromosomes with a panel of somatic cell hybrids. Genomics 9: 721-727. DONIS-KELLER, H., GREEN, P., HELMS, C., CARTINHOUR, S., WEIFFENBACH, B., STEPHENS, K., KEITH, T. P., BORDEN, D. W., SMITH, D. R., LANDER, E. S., BOTSTEXN, D., AKOTS, G., REDIKER, K. S., GRAVIUS, T., BROWN, V. A., RISING, M. B., PARKER, C., POWERS, J. A., WATT, D. E., KAUFFMANN, E. R., BRICKER, A., PHIPPS, P., MULLER-KAHLF,, H., FULTON, T. R., NG, S., SCHUMM, J., BRAMAN, J. C., KNOWLTON, R. G., BARKER, D. F., CROOKS, S. M., LINCOLN, S. E., DALY, M. J., AND ABRAHAMSON, J. (1987). A genetic linkage map of the human genome. Cell 51: 319-337. DUMANSKI, J. P., CARLBOM, E., COLLINS, V. P., AND NORDENSKJ~LD, M. (1987). Deletion mapping of a locus on human chromosome 22 involved in the oncogenesis of meningioma. Proc.

Natl.

Acad.

Sci. USA

84: 9275-9279.

12. DUMANSKI, J. P. (1990). Rapid procedures for the isolation of random chromosome-specific DNA probes from a phage library. Technique 2: 38-42. 13. DUMANSKI, J. P., GEURTS VAN KOSSEL, A. H. M., RUT-I= LEDGE, M., WLADIS, A., SUGAWA, N., COLLINS, V. P., AND NORDENSKJ~LD, M. (1990a). Isolationof anonymouspolymorphic DNA fragments from human chromosome 22q12-qter. Hum. Get&. 84: 219-222. 14. DUMANSKI, J. P., ROULEAU, G. A., NORDENSKJ~LD, M., AND COLLINS, V. P. (199Ob). Molecular genetic analysis of chromosome 22 in 81 cases of meningioma. Cancer Res. 50: 58635867. 15. ELDRIDGE, R. (1981). Central neurofibromatosis with bilateral acoustic neuroma. Zn “Advances in Neurology” (V. M. Riccardi, and J. J. Mulvihill, Eda.), Vol. 29, pp. 57-65, Raven Press, New York. 16. FEINBERG, A. P., AND VOGELSTEIN, B. (1984). A technique for radiolabeling DNA restriction endonuclease fragments to high specific activity: Addendum. Anal. B&hem. 137: 266 267. 17. FOURNEY, R. M., DIETRICH, K. D., AUBIN, R. A., AND PATERSON, M. C. (1988). HindIII polymorphism in the human c-sis proto-oncogene. Nucleic Acids Res. ltb 8197. 18. GONZALEZ, F. J., SKODA, R. C., KIMURA, S., UMENO, M., ZANGER, U. M., NEBERT, D. W., GELBOIN, H. V., HARDWICK,

A MAP

OF

22 LOCI

ON

J. P., AND MEmR, U. A. (1988). Characterization of the common genetic defect in humans deficient in debrisoquine metabolism. Nature 331: 442-446. 19.

HEIM, S., AND MITELMAN, A. R. Liss, New York.

20.

HERZOG, R., HOLZMAN, K., AND BLIN, N. (1990). A TaqI RFLP for the human arylsulfatase A gene. Nucleic Acids Res. 18: 6746. HOGG, D., GORIN, M., HEINZMAN, C., ZOLLMAN, S., MOHANDAS, T., KLISAK, I., SPARKES, R., BREITMAN, M., TSUI, L., AND HORWITZ, J. (1987). Nucleotide sequence for the cDNA of the bovine beta B2 crystallin and assignment of the orthologous human locus to chromosome 22. Eye Res. 6: 13351342. JULIER, C., LATHROP, M., REGHIS, A., SZAJNERT, M.-F., LALOUEL, J.-M., AND KAPLAN, J.-C. (1988). A linkage and physical map of chromosome 22, and some applications to gene mapping. Am. J. Hum. Genet. 42: 297-308.

21.

22.

F. (1987).

“Cancer

KAPLAN, J.-C., AURIAS, A., JULIER, C., PIUEUR, M., AND SZAJNERT, M.-F. (1987). Human chromosome 22. J. Med. Genet. 24: 65-78.

24.

KAPLAN, J.-C., AND EMANUEL, B. S. (1989). Report committee on the genetic constitution of chromosome togenet. Cell Genet. 51: 372-383.

25.

KRAPCHO, K., NAKAMURA, Y., FUJIMOTO, E., EIDRIDGE, R., LEPPERT, M., O’CONNELL, P., LATHROP, G. M., LALOUEL, J.-M., AND WHITE, R. (1988). Isolation and mapping of a polymorphic DNA sequence (pEFZ31) on chromosome 22 (D22S32). Nucleic Acids Res. 16: 5221.

21.

28.

29.

30.

31.

32.

33.

34.

35.

36.

37.

38.

LATHROP, G. M., LALOUEL, J.-M., JULIER, C., AND OTT, J. (1985). Multilocus linkage analysis in humans: Detection of linkage and estimation of recombination. Am. J. Hum. Genet. 37: 482-498. LATHROP, G. M., LALOUEL, J.-M., AND WHITE, R. (1986). Constructions of human linkage maps: Likelihood calculations for multilocus linkage analysis. Genet. Epidemiol. 3: 3952. Lore, I. T., AND RICHARDSON, E. P. (1981). Neuropathological findings and the biology of neurofibromatosis. In “Advances in Neurology” (V. M. Riccardi and J. J. Mulvihill, Eds.), Vol. 29, pp. 23-32. Raven Press, New York. MCDERMID, H. E., DUNCAN, A. M. V., BRASCH, K. R., BURN, J., HOLDEN, J. J. A., MAGENIS, E., SHEEHY, R., BURN, J., KARDON, N., SCHINZEL, A., TESHIMA, I., AND WHITE, B. N. (1986). Characterization of the supernumerary chromosome in Cat Eye Syndrome. Science 232: 646-648. MCKUSICK, V. A. (1987). The morbid anatomy of the human genome. A review of gene mapping in clinical medicine. Part III. Chromosome 13-X, inclusive. Medicine 66: 237-296. NAKAMUFU, Y., LATHROP, M., O’CONNELL, P., LEPPERT, M., BARKER, D., WRIGHT, E., SKOLNICK, M., KONDOLEON, S., LITT, M., LALOUEL, J-M., AND WHITE, R. (1988). A mapped set of DNA markers for human chromosome 17. Genomics 2: 302-309. O’CONNELL, P., LATHROP, G. M., LAW, M., LEPPERT, M., NAKAMURA, Y., HOFF, M., KUMLIN, E., THOMAS, W., ELSNER, T., BALLARD, L., GOODMAN, P., AZEIN, E., SADLER, J. E., CAI, G. Y., LALOUEL, J-M., ANIJ WHITE, R. (1987). A primary genetic linkage map for human chromosome 12. Gecomics 1: 93-102. RATNER, L., JOSEPHS, S. F., JARRET, R., REITZ, M. S., AND WONG-STAAL, F. (1985). Nucleotide sequence of human c-sis

719

22 growth

fac-

RF.ED, K. C., AND MANN, D. A. (1985). Rapid transfer of DNA from agarose gels to nylon membranes. Nucleic Acids Res. 13: 7207-7221. R.OSCHMANN, E., ASSUM, G., AND FINK, T. (1987). RFLP detected with a 5’-bcr-gene-sequence. Nucleic Acids Res. 16: 1583. ROULEAU, G. A., HAINES, J., BAZANOWSKI, A., COLELUCROWLEY, A., TROFATTER, J. A., WEXLER, N. S., CONNEALLY, P. M., AND GUSELLA, J. F. (1989). A genetic linkage map of the long arm of chromosome 22. Genomics 4: l-6. ROULEAU, G. A., SEIZINGER, B. R., WERTELF.CKI, W., HAINES, J. L., SUPERNJMU, D. W., MARTUZA, R. L., AND GuSELLA, J. F. (1990). Flanking markers bracket the neurofibromatosis type 2 (NF2) gene on chromosome 22. Am. J. Hum. Genet. 46: 323-328.

39.

SEIZINGER, B. R., MARTUZA, R. L., AND GUSELLA, J. F. (1986). Loss of genes on chromosome 22 in tumorigenesis of human acoustic neuroma. Nature 322: 644-647.

40.

SEIZINGER, B. R., ROULEAU, G., O-s, L. J., LANE, A. H., ST. GEORGE-HYSLOP, P., HUSON, S., GUSELLA, J. F., AND MARTUZA, R. L. (1987). Common pathogenetic mechanism for three tumor types in bilateral acoustic neurofibromatosis. Science 236: 317-319.

41.

SKODA, R. C., GONZALEZ, F. J., DEMIERRE, A., AND MEYER, U. A. (1988). Two mutant alleles of the human cytochrome P-450dbl gene (P450C2Dl) associated with genetically deficient metabolism of debrisoquine and other drugs. Proc. Natl. Acad. Sci. USA 85: 5240-5243.

42.

TURC-CAREL, C., PHILIP, I., BERGEX, M., PHILIP, LENOIR, G. M. (1983). Chromosomal translocations ing’s sarcoma. N. Engl. J. Med. 309: 497-498.

43.

VIGFUSSON, N. V., ALLEN, L. J., PHILLIPS, J. H., ALSCHIBAJA, T., AND RICHES, W. G. (1986). A neuroendocrine tumor of the small interstine with a karyotype of 46, XY, t(11;22). Cancer Genet. Cytogenet 22: 211-218. WELLER, P., JEFTREYS, A. J., WILSON, V., AND BLANCHETOT, A. (1984). Organisation of the human myoglobin gene. EMBO J. 3: 439-446.

of the 22. Cy-

LATHROP, G. M., LALOUEL, J.-M., JULIER, C., AND OTT, J. (1984). Strategies for multilocus linkage analysis in humans. Proc. Natl. Acad. Sci. USA 81: 3443-3446.

CHROMOSOME

cDNA clones with homology to platelet-derived tor. Nucleic Acids Res. 13: 5007-5018.

Cytogenetics,”

23.

26.

HUMAN

44.

T., AND in Ew-

45.

WHANG-PENG, J., FRETER, C. E., KNUTSEN, T., NANFRO, J. J., AND GAZDAR, A. (1987). Translocation t(11;22) in esthesioneuroblastoma. Cancer Genet. Cytogenet. 29: 155-157.

46.

WHITE, R., LEPPERT, M., BISHOP, T., BARKER, D., BERKOWITZ, J., BROWN, C., CALLAHAN, P., HOLM, T., AND JEROMINSKI, L. (1985). Construction of linkage maps with DNA markers for human chromosomes. Nature 313: 101-105. WHITE, R., LALOLJEL, J.-M., NAKAM~R.A, Y., DONIS-KELLER, H., GREEN, P., BOWDEN, D., MATHEW, C. G. P., EASTON, D. F., ROBSON, E. B., MORTON, N. E., GUSELLA, J. F., HAINES, J. L., RETIEF, A. E., KIDD, K. K., MURRAY, J. C., LATHROP, G. M., AND CANN, H. M. (1990). The CEPH consortium primary linkage map of human chromosome 10. Genomics 6: 393-412. ZANKL, H., AND ZANG, K. D. (1980). Correlations between clinical and cytogenetical data in 180 human meningiomas. Cancer Genet. Cytogenet. 1: 351-356. ZHANG, F. R., AURIAS, A., DELA~RE, O., STERN, H., BENITEZ, J., ROULEAU, G., AND THOMAS, G. (1990a). Mapping of human chromosome 22 by in situ hybridization. Genomics 7: 319-324. ZHANG, F. R., DELATTRE, O., ROULEAU, G., COUTURIER, J., LEFRANCOIS, D., THOMAS, G., AND AURAS, G. (199Ob). The neuroepithelioma breakpoint on chromosome 22 is proximal to the meningioma locus. Genomics 6: 174-177.

47.

48.

49.

50.