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Proc. NatI. Acad. Sci. USA Vol. 85, pp. 8563-8567, November 1988 Genetics Construction of a genetic map of 17 by use of chromosome-mediated transfer (in situ hybridization/acute promyelocytic leukemia/somatic-celi hybrid/ gene/gene mapping) WEIMING XU*, PATRICIA A. GORMANt, SUSAN H. RIDERt, PHILIP J. HEDGE*, GRAHAM MOORE*t, CATRIN PRICHARD§¶, DENISE SHEERt, AND ELLEN SOLOMON*II *Somatic Genetics Laboratory, tHuman Laboratory, and §Human Molecular Genetics Laboratory, Imperial Cancer Research Fund, London WC2A 3PX, United Kingdom Communicated by Walter F. Bodmer, July 28, 1988 (receivedfor review May 13, 1988)

ABSTRACT We used somatic-cell hybrids, containing as used this panel of transfectants to localize most of the their only human genetic contribution part or all of chromo- published gene markers and sequences on . some 17, as donors for chromosome-mediated gene transfer. A By analyzing cotransfection frequencies, we were also able to total of 54 independent transfectant clones were isolated and define groups of loci that are closely located on chromosome analyzed by use of probes or isoenzymes for >20 loci located on 17. Then, by combining CMGT data with results from a panel chromosome 17. By combining the data from this chromosome- of hybrids with well-defined breaks on chromosome 17 and mediated gene transfer transfectant panel, conventional so- from in situ hybridization, we constructed a detailed map of matic-cell hybrids containing well-defined breaks on chromo- the long arm ofchromosome 17 and a less detailed map ofthe some 17, and in situ hybridization, we propose the following short arm. We found CMGT to be particularly useful in the order for these loci: pter-(TP53-RNP2-DJ7S1)-(MYH2- analysis of groups of closely linked markers, such as GALK, MYHl)-D17Z]-CRYB1-(ERBAI-GCSF-NGL)-acute promye- TKI, GHC, UMPH, and GAA, which could not easily be locytic leukemia breakpoint-RNU2-HOX2-(NGFR-COLIAI- resolved by other means. Our results also confirm that (i) MPO)-GAA-UMPH-GHC-TKI-GALK-qter. Using chromo- although substantial lengths of DNA may be transferred some-mediated gene transfer, we have also regionally localized intact (8, 9), interstitial deletions do frequently occur; (ii) the random probes D17S6 to D17S19 on chromosome 17. multiple fragments of transfected can be found in the same clone; and (iii) there is also selection for centro- Structural abnormalities of chromosome 17 are associated meric sequences (8, 9). with important clinical disorders. For example, the leukemic In addition, during the course of this study we obtained cells of nearly all individuals with acute promyelocytic some transfectants that contained only the small regions of leukemia (APL, FAB classification M3) are characterized by chromosome 17 particularly relevant to the study ofAPL and a reciprocal translocation involving the long arms of chro- von Recklinghausen . mosomes 15 and 17 t(15;17)(q22;qll.2-12) (1). Deletions of chromosome 17 have been associated with mental retardation MATERIALS AND METHODS syndromes such as Miller-Dieker syndrome [del(17)pl3] (2) and Smith-Magenis syndrome [del(17)pll] (3,4). In addition, Culture Conditions. The cells were cultured either in RPMI several causing common genetic diseases were shown 1640 or in Dulbecco's minimal essential medium supple- to be located on chromosome 17. For example, the domi- mented with 10% fetal bovine serum. PCTBA1.8, PJT2/A1, nantly inherited disorder von Recklinghausen neurofibroma- and all CMGT transfectants, PLT, KLT, and TLT were tosis was linked to markers in the pericentromeric region of maintained in HAT medium (100 ,uM hypoxanthine/10 ,M chromosome 17 (5). Loss of heterozygosity of chromosome methotrexate/10 ,M thymidine), which selects for thymidine 17p markers was also recently reported in the progression of kinase. The hybrids GPT17.3.2K41 and TRID62 were main- colorectal carcinomas (6). tained in MX medium [mycophenolic acid (25 ,ug/ml)/xan- Construction of genetic maps is central to the study of thine (250 ,g/ml)]. Back selection of CMGT transfectants . Physical maps can be used as a basis for the with 5-bromo-2-deoxyuridine was done by the addition of 5- generation ofevenly spaced restriction fragment-length poly- bromo-2-deoxyuridine (50 ,g/ml) to the medium, thus se- morphisms along . These restriction fragment- lecting for those CMGT transfectants that have lost the TKI length polymorphisms may then allow the demonstration of . This medium was replenished every 3 days for 2-3 positive linkage between human genetic diseases and the weeks. Resulting colonies were maintained in RPMI 1640/ polymorphic DNA markers. Here we report the use of 10% fetal bovine serum and were denoted by the letter B after chromosome-mediated gene transfer (CMGT) in the genera- the name of the original transfectant. tion of a physical map of chromosome 17. CMGT is an Cytogenetic Analysis. Chromosomes from human lympho- established method of constructing hybrid cells containing cytes, hybrids, and CMGT transfectants were all prepared as subchromosomal fragments of a selected donor chromosome described (10). Additional methods are described in the figure (for reviews, see refs. 7 and 8). The generation of transfec- and table legends. tants by CMGT for chromosome 17 has been helped by the Southern Blots and Hybridization. Twenty micrograms of presence of the selectable marker thymidine kinase (TK) restricted DNA was separated on 0.7% agarose gels by located on the long arm. This technique has enabled us to generate a panel of more than 50 transfectants containing Abbreviations: CMGT, chromosome-mediated gene transfer; APL, different transgenomic fragments of chromosome 17. We acute promyelocytic leukemia. tPresent address: Institute of Plant Science Research, Cambridge, United Kingdom. The publication costs ofthis article were defrayed in part by page charge VPresent address: Department of Physiology, University of Califor- payment. This article must therefore be hereby marked "advertisement" nia Medical Center, San Francisco, CA 94143. in accordance with 18 U.S.C. §1734 solely to indicate this fact. I1To whom reprint requests should be addressed. 8563 Downloaded by guest on September 24, 2021 8564 Genetics: Xu et al. Proc. Natl. Acad. Sci. USA 85 (1988)

electrophoresis and transferred to Hybond-N nylon mem- were localized on bands 17q24-+q25 (Fig. 3A). For the branes (Amersham). DNA hybridization probes were pre- GM0271 cell line, 175 grains were scored on 80 metaphase pared by excising the inserts from the cloning vectors with spreads; of the 17 grains present on the normal chromosome the appropriate restriction endonuclease and separating them 17, 12 (71%) were present on bands 17q24-*q25. Of the 9 on a low-melting-agarose gel. The purified insert was radio- grains present on the 19p+ derivative chromosome, 7 (78%) labeled with [32P]dCTP (Amersham) by the random-priming were located in the 17q23--q25 region of chromosome 17 method (29). Hybridizations were done as described (30). (Fig. 3E). GHC. Regional localization of the GHC genes was ob- RESULTS tained by analysis of chromosome 17 in 28 metaphase spreads; of the 38 grains present, 27 (71%) were located on Southern Blot and Isoenzyme Analysis. Southern blots (11) bands 17q23--+q24 (Fig. 3B). ofrestricted DNA from each ofthe CMGT transfectants were HOX2. Regional localization of the HOX2 locus was hybridized with probes for the loci indicated in Table 1. The obtained by analysis of chromosome 17 in 40 metaphase presence or absence ofthe human specific bands detected by spreads; of the 50 grains present, 28 (56%) were located on these probes corresponds to the presence or absence ofthese bands 17q21-+q22 (Fig. 3C). regions of chromosome 17 in the transfectants. This infor- CRYB1. Chromosomes from 70 metaphase spreads of mation is summarized together with the results of the peripheral blood lymphocytes were analyzed; 149 grains isoenzyme analysis in Fig. 1. The frequencies with which were scored, ofwhich 17 were found on chromosome 17, and markers are cotransferred with the selected gene TKI are 12 (71%) of these were located on bands 17q11.1-*q12 (Fig. indicated in Fig. 2. 3D). Lotalization of Probes by in Situ Hybridiiation. TK1. This Mapping of Loci on Chromosome 17. In constructing a map locus was mapped with chromosomes from both peripheral of human chromosome 17 we assumed the following: (i) the blood lymphocytes and the cell line GM0271, which has the proximity of markers to the selected locus TKI would be translocation t(17;19)(q23;p13.3) (36). Chromosomes from indicated by the frequencies with which they were found in 100 metaphase spreads ofperipheral blood lymphocytes were the transfectants, and (ii) those markers with similar cotrans- analyzed. A total of 218 silver grains were scored, of which fection frequencies with TKI and that are generally found in 28 were found on chromosome 17, and of these, 18 (64%) the same transfectants would likely be located close together Table 1. Loci used to characterize the CMGT panel of transfectants Previous* Gene Name Probe localization Reference TPS3 Tumor p53-102 17p13 12 RNP2 Large subunit RNA polymerase pHRpll5.5 17p12-+p13 13 D17S6 DNA single copy cosH17.1 17pter-.p13.3 14 D17S7 DNA single copy cosH17.2 17cen--pter 14 D17SI DNA single copy p12-2 17pter-+p13 15 MYH2 heavy polypeptide, skeletal muscle, 2 adult 2 p10-3 17pter-+pll 16 MYHI Myosin heavy polypeptide, skeletal muscle, 1 adult 1 p2-3 17pter-*pll 16 DJ7ZI DNA multiple copy p17H8 17cen 17 CRYBI 13 crystallin pCU14A1 17q21 18 ERBAI Erythroblastic leukemia viral oncogene homology pHE-Al 17qll-.ql2.21 19 GCSF Granulocyte colony-stimulating factor pBRG-4 17qll-+ql2.21 20 NGLt Neuro/glioblastoma Amprobet 17q21-+q22 2 oncogene homology erbB2 RNU2 RNA, U2 small nuclear pML2d 17q21-.q22 21 D17S17 DNA single copy cosH17.11 17q22--qter 14 D17S18 DNA single copy cosH17.12 17q22--qter 14 D17S19 DNA single copy cosH17.13 17q22--qter 14 HOX2 region 2 pHU-1 17qll-.q22 22 NGFR Nerve growth factor receptor p104 17ql2-+q22 23 COLIAI Collagen al(I) pCG103 17q21.31-+q22.5 24 MPO pMP23 17q11.12-*q24 25 GAA Acid a-glucosidase Enzyme 17q23 26 UMPH Uridine 5'-monophosphate phosphohydrolase Enzyme 17q 26 GHC complex-homologous genes: pBGHXH01 17q22-*q24 27 GHl-CSPHl-CSHl-GH2-CSHP2 TKI Thymidine kinase pHTK2 17q21-+q22 28 D17S9 DNA single copy GERBA1 17qll 14 D17SIO DNA single copy cosH17.5 17qll-+q22 14 D17SII DNA single copy cosH17.6 17qll- q22 14 D17S12 DNA single copy cosH17.7 17q11- q22 14 D17S13 DNA single copy cosH17.8 17qll-.q22 14 D17S14 DNA single copy cosH17.9 i7qll-+q22 14 D17S16 DNA single copy cosH17.10 17qll- q22 14 GALK Enzyme 17q21-.q22 26 Isoenzyme analyses were all done with starch gel electrophoresis according to Harris and Hopkinson (26). *Prior localizations of the above loci are taken from refs. 31 and 32. tNGL is now officially known as ERBB2. tAmersham International. Downloaded by guest on September 24, 2021 Genetics: Xu et al. Proc. Natl. Acad. Sci. USA 85 (1988) 8565

E _tv Ngtv tTsted t MPositive E]Negative J3Not.Tested UnstabLe FIG. 1. Schematic representation of the presence or absence of chromosome 17 markers in the panel of transfectants and somatic-cell hybrids. Markers are displayed in their approximate order and position on chromosome 17 as determined by this study. DCI, donor cell line: P, PCTBA1.8; K, GPT17.3.2K41; and T, TRID62. q+, 15q+ hybrid. The CMGT protocol has been described in detail (33). Three human/mouse interspecific somatic-cell hybrids were used as chromosome donors. PCTBA1.8 is a mouse 3T3TK-/human somatic-cell hybrid (34) containing a chromosome 17 as its only human component. GPT17.3.2K41 is a mouse teratocarcinoma/human somatic-cell hybrid (35) also containing chromosome 17 as its only human component. TRID62 is a mouse/human somatic-cell hybrid containing as its only human component the long arm of chromosome 17 (17qll.2--qter) (35). All resultant CMGT transfectants derived from PCTBA1.8, GPT17.3.2K41, and TRID62 carry the prefix PLT-, KLT-, and TLT-, respectively. The mouse cell line LMTK- was used as the thymidine kinase-deficient recipient cell line for all the CMGT experiments. The PJT2/A1 hybrid, sometimes referred to as the 15q+ hybrid, contains the APL translocation chromosome with the region of 17 below the breakpoint (17qll.2-.qter) (10). on the physical map. By combining the data from CMGT, In situ hybridization has localized the thymidine kinase somatic-cell hybrids, and in situ hybridization, we were able gene to the bands 17q24-*q25. This position is more telome- to identify six major regions of chromosome 17 covering the ric than that previously reported (17q21--q22) (40). It is, long arm through the to the short arm. however, consistent with the more recent findings of others, Region 1. This region includes the selectable marker TKI where TKI was mapped to 17q23-*qter with a hybrid derived and four other genes (GALK, GHC, UMPH, and GAA) that from GM0271, by showing that thymidine kinase activity have cotransfection frequencies with TKI of >50%. GHC has remained only in the presence of the derivative 19 chromo- the highest cotransfection frequency with TKI and can some (41). The GHC locus has been assigned to chromosome therefore be assumed to map closest to the TKI locus. 17 and localized by in situ hybridization to chromosome Because the transfectant PLT7 only has TKJ and GALK and bands 17q22--q24 (42). Here we report confirmation of this transfectant PLT4 only has TKI and GHC, it is probable that assignment and have further localized it to 17q23->q24. This GHC and GALK are located on either side of TKI. UMPH regional localization of GHC is consistent with the GHC cotransfects with the GHC genes with a high frequency, locus being closer to the centromere than TKI. Region 1 indicating that it lies adjacent to GHC. probably spans 17q23-}q25 with the gene order being cen- tromere (cen)-GAA-UMPH-GHC-TKI-GALK-qter. We have also used this CMGT transfectant panel with several other probes isolated from a human chromosome-17- only library. D1759, D17SJO, D17SII, D17S12, D17S13, 80- D17S14, and Dl 7S16 (14) were detected on the transfectant 0- PLT8 and have therefore been assigned to region 1, 17q23-*qter. D17S9 and DJ7510 have cotransfer frequencies with TKI of >90%, indicating very close proximity to TKI. It should be noted that although D17S9 has also been localized by in situ hybridization to 17q23-,qter, it also o cross-hybridizes to sequences just below the APL break- point. Region 2. The loci HOX2, NGFR, COLIAJ, and MPO C.~~~~~~~~~~~~~~M C compose the second group, as they are commonly found together in the same transfectants. The MPO probe has only Tac40 SIS< SMDtEm- Q( been used with a more limited CMGT transfectant panel. However, MPO is always present with COLIAJ, indicating the close proximity of these loci; for example, they are both present in the transfectants PLT6, TLT2, TLT8, TLT10, and TLT20. COLIAJ is probably closer to TKI than either NGFR or HOX2, as it has a higher cotransfection frequency with TKI. HOX2 is likely to be located more proximal to the Gene centromere than the other loci in region 2, as it is not cotransferred with them in the transfectants PLT6, PLT15, FIG. 2. Overall gene frequencies of chromosome 17 markers in and TLT8. the PLT and KLT transfectant series. Frequencies are expressed as KLT13, However, HOX2 is found in KLT3, a percentage of the total number of transfectants that acquired the TLT10, and TLT20, suggesting that it is located in this region. human thymidine kinase gene. TLT transfectants are not included, This hypothesis is substantiated by the loss ofHOX2 together as the donor chromosome for this series was not a complete with MPO, COLJAJ, and NGFR in the transfectant PLT20, chromosome 17. which is deleted for this region. The close proximity of Downloaded by guest on September 24, 2021 8566 Genetics: Xu et al. Proc. Natl. Acad. Sci. USA 85 (1988)

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0 *000 25 @000000.000000000. 17 17 17 E

17 17q 19 19 p

FIG. 3. In situ hybridization to localize the loci TKI, GHC, HOX2, and CR YB). The distribution ofgrains on chromosome 17 is represented schematically for the following loci: TK) (A), GHC (B), HOX2 (C), and CRYB) (D). (E) A more precise localization of TKI was done using the human fibroblast cell line GM0271, which is characterized by a reciprocal translocation t(17;19)(q23;pl3.3) (37). Before in situ hybridization, cells were treated with 5-bromo-2-deoxyuridine (37). In situ hybridizations were done with the method of Harper and Saunders (38). The probes for the TKI, GHC, HOX2, and CR YBI loci were all radiolabeled using oligoprimers and [3H]deoxyribonucleotides to a specific activity of 1 x 107-1 x 108 cpm per jzg of DNA. In situ hybridizations were done overnight at 37°C with a final probe concentration of 0.1-0.02 ,tg/ml. Slides were then washed at 39°C, dehydrated, and dipped in Ilford K-5 emulsion. All slides were developed after 1-3 weeks and then G-banded (39); the slides were also stained with Wright's stain.

COLIAI, MPO, and NGFR is substantiated by the loss of This positioning is inconsistent with our more proximal these three genes, together with the region-1 markers, in the localization of CR YBI at 17qll.1-+ql2. back-selected transfectant PLT6.B. The transfectant PLT6 Region 6. Many transfectants containing the centromere of contains two fragments of chromosome 17 integrated into chromosome 17 also contain markers from the short arm. The separate regions of the mouse . Selection with 5- probes from the short arm are rather far from the TKI locus, bromo-2-deoxyuridine results in the loss of the fragment but we do see these genes in transfectants such as PLT6, containing the thymidine kinase gene and, consequently, loss PLT15, PLT20, KLT3, and KLT12. TP53, RNP2, and D17S5 of the other loci on this fragment. are always associated with each other, as are MYHI and In situ hybridization confirms the localization of HOX2 to MYH2 (except in transfectant PLT6). We therefore tenta- 17q21-*q22 (43), and COLIA) has been previously mapped tively divide this region into these two subregions. to 17q21-*q22 (44). It therefore appears that this second D17S6 and D17S7 (14) are also located on the short arm of group is located in the region 17q21-+q22, most probably in chromosome 17, 17pter--cen. the order cen-HOX2-(NGFR-COLIAI-MPO)-qter. Region 3. The gene for small nuclear RNA U2 (RNU2) maps below the APL breakpoint. Although this gene has a DISCUSSION similar cotransfection frequency with TKI to the region-2 We describe our results using CMGT methods to generate loci, RNU2 does not belong to the same localization group, transfectants containing different fragments of human chro- as it is absent from cell lines KLT3 and TLT10 and is retained mosome 17. Originally CMGT was limited to the availability in cell line PLT6.B, which has lost regions 1 and 2. RNU2 has of endogenous dominantly acting selectable markers such as therefore been placed in a separate region, which lies closer TKI and HPRT, which enabled selection for chromosomes 17 to the centromere than 2. The loci region DI7S)7, DI7S18, and X, respectively. The technique has now been extended and have been localized to below DI7SI9 (14) 17q11.2-+q22, of the genome the use of other forms of the APL breakpoint and above the region covered by cell line to other regions by selection, such as selection of human transforming genes- PLT8, but as yet these loci have not been further localized to e.g., HRASI (45)-or by cotransfecting with a selectable either region 2 or 3. (33). In our study, by selecting for Region 4. Using the hybrids TRID62 and PJT2/A1, we marker-e.g., pSV2neo have mapped the loci ERBAI, GCSF, and NGL close to but the thymidine kinase gene we obtained a panel of >50 a We below the centromere on the long arm ofchromosome 17 and transfectants covering large region of chromosome 17. above the APL breakpoint 17q11.1- q12. The GCSF probe observed that our transfectants frequently have some inter- has only been used with a more limited CMGT transfectant stitial deletions. Sometimes these deletions cover one rather panel. However, GCSF is always present with ERBAI-for large region, but many transfectants are deleted for several example, in the transfectants KLT8, KLT12, and TLT2. different short regions, clearly showing serious drawbacks to NGL is also present in transfectants KLT8 and TLT2 but not using CMGT alone for gene mapping. However, when we KLT12. As KLT12 also contains the centromeric sequence combined our CMGT data with data from both a chromosome D17ZI, it is likely that ERBAI and GCSF are located closer 17 hybrid panel that contained well-characterized constitu- to the centromere than NGL, the order being as follows: cen- tional rearrangements of human chromosome 17 and in situ (ERBAI-GCSF)-NGL-qter. hybridization, the deleted regions could be detected and the Region 5. The centromeric region is represented by D17ZI genes could be mapped in finer-detail than would have been and CR YBI. Interestingly, the human centromere has a possible using any one of these methods. Using these three relatively high cotransfection frequency with TKI compared methods, we constructed the following order of markers on with adjacent loci, suggesting that there is selection for these chromosome 17: pter-(TP53-RNP2-D17SI)-(MYH2- sequences. We found that all the transfectants that contained MYHI)-D)7Z)-CRYBI-(ERBAI-GCSF-NGL)-APL break- the locus D)7ZI also retained the CR YBI locus. In situ point-RN U2-HOX2-(NGFR-COLIAl-MPO)-GAA- hybridization had previously mapped CRYBI to 17q21 (18). UMPH-GHC-TKI-GALK-qter. Downloaded by guest on September 24, 2021 Genetics: Xu et al. Proc. Natl. Acad. Sci. USA 85 (1988) 8567

Recently some family studies have demonstrated the son, J. R. & Solomon, E. (1983) Proc. Natl. Acad. Sci. USA 80, genetic distance between some of the markers used in this 5007-5011. 11. Southern, E. M. (1975) J. Mol. Biol. 98, 503-517. study. These studies show that DI7ZJ and COLIAI are about 12. Matlashewski, G., Lamb, P., Pim, D., Peacock, J., Crawford, L. & 20 centimorgans apart, GHC and DI7ZJ are about 28 centi- Benchimol, S. (1984) EMBO J. 3, 3257-3262. morgans apart, and NGFR and HOX2 are closely linked (46, 13. Cannizzard, L. A., Emanuel, B. S., Weinman, R. & Cho, K. Y. 47)-results that are consistent with our data. (1986) Am. J. Hum. Genet. 38, 812-816. Possibly the most useful aspect of CMGT is that it 14. Harper, M. E., Barrera-Saldana, H. A. & Saunders, G. F. (1982) Am. J. Hum. Genet. 34, 227-234. generates hybrid cell lines that are enriched for certain small 15. Naylor, S. L., Sakaguchi, A. Y., Barker, D., White, R. & Shows, regions ofthe human chromosome. This result is ofparticular T. B. (1984) Proc. NatI. Acad. Sci. USA 81, 2447-2451. value when the genes of interest are located near the 16. Leinwand, L. A., Fournier, R. E. K., Nadal-Ginard, B. & Shows, selectable marker. CMGT can also be used when the select- T. B. (1983) Science 221, 766-769. able marker is distant from the of interest due to the 17. Waye, J. S. & Willard, H. F. (1986) Mol. Cell. Biol. 6, 3156-3165. region 18. Law, M. L., Cai, G.-Y., Hartz, J., Kao, F.-T., Hogg, D., Breitman, high frequency of interstitial deletions, which allows frag- M. L. & Tsui, L.-C. (1984) Cytogenet. Cell Genet. 42, 202-207. ments normally distant from the selectable marker to be 19. Jansson, M., Philipson, L. & Vennstrom, B. (1983) EMBOJ. 2, 561- transferred on relatively short pieces ofthe chromosome. For 565. example, the transfectant KLT8 contains the loci from 20. Shigekazu, N., Masayuki, T., Shigetaka, A., Yoshito, K., Tatsumi, 1 and but not those from 2 and and could Y., Osami, Y., Yuichi, H., Naoki, K., Masayoshi, O., Hitoshi, N. regions 4, regions 3, & Masayoshi, 0. (1986) Nature (London) 319, 415-418. therefore be useful in the study of von Recklinghausen 21. Lindgren, V., Ares, M., Jr., Weiner, A. M. & Francke, U. (1985) neurofibromatosis and APL. In addition, the transfectant Nature (London) 314, 115-116. PLT6.B, which has only :'13% of chromosome 17 according 22. Levine, M., Rubin, G. M. & Tjian, R. (1984) Cell 38, 667-673. to our DNA dot blot analysis (W.X., unpublished data) and 23. Huebner, K., Isobe, M., Chao, M., Bothwell, M., Ross, A. H., which has been deleted for most the could Finan, J., Hoxie, J. A., Sahgal, A., Buck, C. R., Lanahan, A., of long arm, also Nowell, P. C., Koprowski, H. & Croce, C. M. (1986) Proc. Natl. be of value for this purpose. Acad. Sci. USA 83, 1403-1407. While this manuscript was in preparation, another group 24. Barsh, G. S., Rousch, C. L. & Gelinas, R. E. (1984) J. Biol. Chem. published data on a regional mapping panel for human 259, 14906-14913. chromosome 17 (40). Their approach was to construct inter- 25. Chang, K. S., Trujillo, J. M., Cook, R. G. & Stass, S. A. (1986) somatic-cell from human Blood 68, 1411-1414. specific hybrids cells that had 26. Harris, H. & Hopkinson, D. A. (1976) Handbook of Enzyme well-defined breaks on chromosome 17. Our approach was Electrophoresis in Human Genetics (North-Holland, Amsterdam). different in that it was largely based on data from a panel of 27. Morello, D., Moore, G., Salmon, A. M., Yaniv, M. & Babinet, C. CMGT transfectants with some somatic-cell hybrids. There (1986) EMBO J. 5, 1877-1883. are no inconsistencies for the localization of loci on chromo- 28. Lau, Y.-F. & Kan, Y. W. (1984) Proc. Natl. Acad. Sci. USA 81, some 414-418. 17 in both studies. Our data give a more detailed map 29. Feinberg, A. P. & Vogelstein, B. (1984) Anal. Biochem. 137, 266- of the long arm of chromosome 17, and their data supply a 267. more detailed map ofthe short arm ofthe same chromosome. 30. Solomon, E., Hiorns, L. R., Spurr, N., Kurkinen, M., Barlow, D., Hogan, B. L. M. & Dalgleish, R. (1985) Proc. Natl. Acad. Sci. USA Note Added in Proof. More recent data make the ordering ofERBAI, 82, 3330-3334. GCSF, and NGL ambiguous. 31. Eighth International Workshop on Human Gene Mapping (1985) Cytogenet. Cell Genet. 40 (1-4). We thank the following people for generously providing probes for 32. Ninth International Workshop on Human Gene Mapping (1987) the loci used in this study: G. Matlasheski (TP53), N. Spurr (RNP2, Cytogenet. Cell Genet. 46 (1-4). D17SJ), H. Willard (DJ7ZJ), B. Vennstrom (CERBAI), N. Shige- 33. Pritchard, C. & Goodfellow, P. N. (1986) EMBO J. 5, 979-985. kazu A. 34. Bai, Y., Sheer, D., Hiorns, L., Knowles, R. W. & Tunnacliffe, A. (GCSF), Weiner (RNU2), C. Hauser (HOX2), K. Heubner (1982) Ann. Hum. Genet. 46, 337-347. (NGFR), G. Barsh (COLIAI), K. Chang (MPO), and Y. Kan (TKI). 35. Tunnacliffe, A., Parkar, M., Povey, S., Bengtsson, B. O., Stanley, We thank Sue Povey and Annabel Kearney for the isoenzyme K., Solomon, E. & Goodfellow, P. N. (1983) EMBOJ. 2,1577-1584. analysis. We are grateful to Peter Goodfellow for his advice on 36. National Institute of General Medical Sciences Human Genetic CMGT and his valuable discussions on the manuscript. We also Mutant Cell Repository (1985) Catalog of Cell Lines (Natl. Inst. thank Mrs. P. Miller for her help in the laboratory. Health, Bethesda, MD), NIH Publ. No. (NIH) 85-2011. 37. Wolff, S. & Perry, P. (1974) Chromosoma 48, 341-353. 1. Rowley, J. D., Golomb, H. M., Vardiman, J. W., Fukuhara, S., 38. Harper, M. E. & Saunders, G. F. (1981) Chromosoma 83, 431-439. Dougherty, S. & Potter, D. (1977) Int. J. Cancer 20, 860-872. 39. Zabel, B. U., Naylor, S. L., Sakaguchi, A. Y., Bell, G. 1. & Shows, 2. Stratton, R. 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