Comparative Mapping in the Domestic Cat (Felis catus)

S. J. O'Brien, S. J. Cevario, J. S. Martenson, M. A. Thompson, W. G. Nash, E. Chang, J. A. M. Graves, J. A. Spencer, K.-W. Cho, H. Tsujimoto, and L. A. Lyons

The genetic map of the domestic cat has been developed as a model for studying Downloaded from both feline analogues of human genetic disease and comparative genome organi- zation of mammals. We present here the results of syntenic mapping of 35 based upon concordant occurrence of feline gene homologues with feline chro- mosomes and previously mapped loci in a panel of 41 rodent x cat somatic cell hybrids. These somatic cell hybrids retain rodent and segregate fe- http://jhered.oxfordjournals.org/ line chromosomes, but in different combinations in each hybrid cell line. Thirty- three of the 35 new locus assignments extend and reaffirm conserved segment homologies between the human and cat genomes previously recognized by comparative mapping and zoo-FISH. These results demonstrate the extensive syntenic conservation between the human and feline genomes and extend the fe- line gene map to Include 105 assigned loci.

Genetic mapping of homologous loci in di- ical comparisons (Lyons et aJ. 1994, 1997; verse species reveals that genomic orga- Nash and O'Brien 1982, O'Brien and Nash nization is not a random process (Com- 1982; O'Brien et al. 1993, 1997). More re- at University of California, Davis - Library on September 12, 2012 parative Genome Organization 1996; Co- cently, reciprocal chromosome painting peland et al. 1993; DeBry and Seldin 1996; using individual probes from flow-sorted Nadeau et al. 1995; O'Brien et al. 1988). human and feline metaphase chromo- Blocks of linked genes have been shown somes has demonstrated by direct obser- to be preserved throughout evolution and vation the conservation between the fe- these blocks can exist intact in species as line and organizations diverse as humans and flies. This concept (O'Brien et al. 1997; Wienberg et al., in is best exemplified by the comparison of press). genomic localizations of homologous Pathological analogues of over 30 inher- genes that have been mapped in both hu- ited human diseases have been described mans and mice. Approximately 130 chro- in the domestic cat (Migaki 1982; Nicholas mosomal segments are shown to be con- and Harper 1996; Nicholas et al. 1996). Fe- served when homologous genes are lines are also an excellent model for infec- aligned in the two species [Comparative tious and acquired diseases, specifically Genome Organization 1996; Copeland et feline leukemia virus (Hardy et al. 1980) al. 1993; DeBry and Seldin 1996; Nadeau et and feline immunodeficiency virus (Car- al. 1995; Mouse Genome Database (MGD)]. penter and O'Brien 1995; Pedersen 1993). The conserved segments likely reflect an Feline leukemia is an extensively studied From the Laboratory of Genomlc Diversity, National ancestral genomic organization that has model for virus-induced cancer, while fe- Cancer Institute, Frederick Cancer Research and De- been inherited throughout the evolution line immunodeficiency virus induces feline velopment Center, Frederick, MD 21702-1201 (O'Brien, Cevario, Martenson, Thompson, and Lyons), H&W Cy- of rodents and primates. Similar conclu- AIDS. Extension of the cat gene map will togenetlc Services, Inc., Lovettsvllle, Virginia (Nash), sions were reached when comparison of facilitate disease gene analyses and the fe- the Lombardl Cancer Center, Georgetown University, human and cattle gene maps revealed ex- line genome can serve as the carnivore Washington, D.C. (Chang), the Department of Genetics and Human variation. La Trobe University, Bundoora, tensive conserved syntenic segments representative for genome evolutionary Victoria, Australia (Graves and Spencer), and the De- (Barendse et al. 1994; Bishop et al. 1994; studies. partment of Veterinary Internal Medicine, University of Tokyo, Tokyo, Japan (Tsu)lmoto, Cho). This paper was Ma et al. 1996; Womack and Kata 1995). We are continuing to develop the genet- delivered at a symposium entitled "Interspedes Hy- The feline genome has also proven to ic map of the cat using both type 1 (coding brids In Mammals" In association with the New Zea- land Genetical Society and Australasian Gene Mapping have a genomic organization highly con- gene) and type n (microsatellite markers) Workshop In Dunedln, New Zealand, from November served relative to human. This conserva- loci (Menotti-Raymond and O'Brien 1995; 30-December 1, 1995. tion has been evident by both compara- O'Brien 1991; O'Brien et al. 1993). In the Journal of Heredity 1997^8:408-414; 0022-1503/97/15.00 tive gene mapping and G-banded cytolog- present study, 35 genes are assigned to fe-

408 Table 1. Molecular gene clone* used to track feline homologues In Southern blot analyses

Species of R.E. Mouse/ Gene symbol Clone Vector Insert (Kb) R.E.- origin Hamster Reference/Source

ABU pAOgl pUC19 1.2&ORI Feline Bg/U Blochlmica et Blophysica Acta 824:104-112, 1985 ARAF1 SAraQ81 pUC19 I.6&0RI Mouse Bg/U Mol Cell Biol 6:2655-2662, 1986 CD8A pT8Fl pSP6 1.7£coRI Human BamHl Cell 40:241, 1985 COL1A1 Hf677 pBR322 1.8£coRl Human Bg/1 ATCC #61322 CSF1R EGFR pAEBamRI pBR322 BamHl/EcoW Avian BamHI ATCC #41019 EGR1 EGR1 pUC13 3.1 £coRI Mouse Bg/II Sukhatme (unpublished) F9 pf9 pGEM 1.5 Human PsA Nature 299:178-180, 1982 FES pA08g pAT153 tf/ndUI/£coRI Feline Kpn\ Gene 35:33-43, 1985 FGF1 EGG177 pDH15 2.2 EcoRl Human EcoKi/Bgl II Science 233:541-545, 1986 FGF3 p39A pUC12 0.6 7a<7l Mouse BamHI R. Cardiff FOS p(os-l pBR322 0.9 ft/I Mouse J. Virol 44:674, 19?? FYN pFyn pucl9 1.6 tfincIU/fcoRl ftrl/BamHI ATCC GU pKK36pl pGem3 1.55 ft/1 Human Bgill/Bamrfl Science 236.70-73, 1987 H0XA4 Hox 1.4 pBR322 0.98 Windlll Human 6:oRI Genomics 5:250-258, 1989 pUC EJ6.6 HRAS pUC13 6.6 BamHI Human Sad ATCC #41028 Downloaded from HRASP pBR-NT pBR322 10.7 BamHI Human ATCC #41001 ILIA IL-la pBR322 1 7 EcoRl/HwdlW Human BamVA DA1NIPPON Pharmaceutical CO:LTD IL8 IL8 pucl9 0.5, 1.2&ORI Human EcoN J Exp Med 167:1883, 1988 JUN pHJ 0 9ftrt Human flg/II Bohman KIT phcldt-171 pUC19 1.25 Ssl\ Human ftrt/Bg/I ATCC #59492 KRAS2 pSWll-1 pUC13 0.6, 0.5 ft/I/£coRI Human Sad ATCC #41027 MOS pAB pBR322 0.6 Aval/BamHl Mouse EcofQ G. Vande Woude MYB c-myb pUC18 0.5£coRl Mouse Bg/II PNAS 835010-5014, 1986 http://jhered.oxfordjournals.org/ MYC pHSR-1 pBR322 9.0 EcoRl Human ATCC #41010 MYCN pNB-1-Sub pGem3 1.0£coRI/BamHI Human Bg/D ATCC #41011 NRAS p52C pUC12 1.5£coRl Human Sad ATCC #41030 ore pOTC pUC9 1.3 Rat ft/I Eur J Blochem 143:183-187, 1984 PDGFB pPHS-1 pUC18 1.3 BamHl Feline BglO Gene 3533-43, 1985 PIM1 phplm5R] pyt 3.6 £coRl Human Ssl\ ATCC #59168 ROS1 pROS pBR322 0.8 PvM/EcoRl Avian Hindin/HindlU B. Vogelstein SRC phu-csrc pUC8 \.7 Hindm/EcoRl Human Bgifl Mol Cell Blol 5:831-338, 1985 THRA1 pHE-Al pBR322 3.9 £coRJ Human BamHl/Ssti J Virol 36(2):575-585, 1980 WNT1 pMT2.5 pBR322 2.5 BamHl/firoRI Mouse Ssfi Cell 31:99-109, 1982 YES1 pXEyes pUC19 0.55 £coRJ/ftrt Human ft/I ATCC #57582 at University of California, Davis - Library on September 12, 2012 • Restriction eiuymes used to release the Insert from the vector.

line chromosomes by Southern blot anal- O'Brien and Nash 1982). Hybrid lines were following standard protocols (Modi et al. yses using a panel of somatic cell hybrids characterized by chromosomal G-banding 1987) with the following modifications. between rodent and feline cells (O'Brien and isozyme typing, which has been de- Southern blots were hybridized for 40 h at and Nash 1982). Each hybrid cell line has scribed (Berman et al. 1986; Gilbert et al. 37°C in 50% formamide, 1 M NaCl, 50 mM been karyotyped to identify the retained 1988; Masuda et al. 1991; Nash and O'Brien PIPES (pH 6.8), 200 ng/ml denatured salm- feline chromosomes. Seventy genes have 1982; O'Brien and Nash 1982). The feline on sperm DNA, 10 mM EDTA, and 10X been previously mapped using this same chromosomal constitution of a single pas- Denhardt's solution (0.2% Ficoll, 0.2% po- hybrid panel. Thirty-three of the new gene sage for each hybrid cell line Is presented lyvinylpyrrolidone, and 0.2% bovine se- assignments could be predicted by previ- in Table 2. Genotypes for all passages of rum albumin). The gene-specific insert for ous mapping data that delineated blocks every member of the cell panel are includ- each probe was isolated from the vector of chromosomal segments that are con- ed in the Web site for the Laboratory of using the appropriate restriction served between felines and humans. The Genomic Diversity (http://www.nci.nih.gov/ (Table 1). The inserts were separated new genes increased the genetic map of intra/lgd/lgdpage.html). from the vectors by agarose-gel electro- the cat to 105 loci, which encompasses 18 DNA was purified from the parental and phoresis and the inserts were isolated by of the 19 feline chromosomes (27V = 38), the 42 hybrid cell lines by standard phenol excision from the gels followed by purifi- showing 15 multigene blocks of conserved and chloroform extraction methods (Man- cation with GeneClean (Bio 101). Vector chromosomal segments with humans. iatis et al. 1982). Southern blots of the pa- inserts were radiolabeled by random rental samples were used to determine priming following manufacturer's recom- which enzymes would distinguish hybrid- mendations (Boehringer Mannheim). The Materials and Methods ization patterns that would be diagnostic filters were initially washed in 2x SSC, Molecular clones and probes for the 35 between the cat, the mouse, and the ham- 0.1% SDS for 30 min at 50°C; the final wash genes are described in Table 1. The iden- ster. Each sample (10 ng) was digested was lx SSC for 30 min at 65°C. Washes tity of each probe was verified by confirm- with seven different enzymes—BamHI, were changed every 30 min and wash ing that the expected fragment size from Bglll, EcoRl, Hindm, ft/I, Sst\, and Kpn\— stringencies were increased as required human DNA for each probe was obtained using conditions as recommended by the for each probe. Stringency was increased by a Southern blot analysis. The develop- manufacturer (GIBCO BRL). Digested sam- by increasing temperatures and decreas- ment of the hybrid cell lines has been de- ples were separated by electrophoresis In ing SSC concentration as required to re- scribed (Berman et al. 1986; Gilbert et al. 1.0% agarose gels at 40 V for 18 h. DNA duce background radiation on the filters. 1988; Masuda et al. 1991; O'Brien 1986; blotting and hybridization were performed Blots were exposed to X-ray diagnostic

O'Brien et al • Comparative Gene Mapping in Domestic Cat 409 Table 2. Chromosome constitution of cat x rodent somatic cell hybrid panel ure 1). Each gene produced a hybridiza-

Feline chromosomes tion signal specific for the cat that was a different molecular size(s) than the signals Hybrid Al A2 A3 Bl B2 B3 B4 Cl C2 Dl D2 D3 D4 El E2 E3 Fl F2 X Y produced by the mouse or the hamster 17T1G + + + + + + + + (Figure 1). The chromosome assignment, 17T2F + + + + + + + + range of discordant hybrid frequency, plus 17T3E 100 80 80 80 20 100 20 10 100 30 100 17T4E 35 25 30 50 50 25 60 25 40 60 15 a chi-square test for gene chromosome as- 17T5H + + 60 sociation is presented in Table 3. An av- 17T6D + + + + + erage of 32 hybrid lines were scored to cal- 17T7D + 17T8F + + + culate concordancy and disconcordancy 17T9D 80 10 100 100 frequencies. 17T10C + 100 + 100 17T11D 60 Gene markers were assigned to chro- 17T12G + 60 90 100 + 100 90 100 mosome based on concordance with 17T26D + + G-banded chromosomes, by concordance 17T27D + + + + 17T28E 20 10 35 5 40 50 60 100 10 with gene markers previously assigned to 17T29D + + + + + + + + that chromosome, and by high discor- Downloaded from 17T30C + + + + dance with other chromosomes and gene 17T33E 100 80 100 100 100 100 17T34D 100 10 5 35 100 markers. Thus the genes CD8A, ILIA, 17T35F 50 50 40 50 40 30 20 5 80 30 80 40 MYCN, and SRC were assigned to feline 17T36E 50 70 50 60 70 40 20 15 70 40 5 70 50 17T37D 40 + 100 40 chromosome A3 with four supportive

49C1E 100 + 100 markers each, and FGF3 was assigned to http://jhered.oxfordjournals.org/ 49C3E + + + + + + + + feline chromosome Dl with three support- 49C4B + + + + + + + + + + + + 49C5C 65 18 47 18 6 12 53 24 18 82 ive markers (Table 3). These four genes 49C6D + + + + also displayed high discordancy frequen- 49C7D 100 100 100 cies with chromosomes and genes from 49C9C 64 9 82 55 18 18 49C10B + + + + + + the other feline chromosomes. For exam- 49C11C 14 50 7 71 21 7 86 7 79 86 ple, CD8A maps to feline chromosome A3 49C12D 20 20 67 7 13 33 67 7 67 49C13F 90 80 70 100 100 100 40 + + 30 100 100 concordant with four gene markers (ADA, 49C14F + + + ACPI, MDH1, and ITPA). The discordancy 49C15E 60 80 60 80 80 50 100 80 80 40 100 100 of CD8A versus chromosome A3 markers 49C16G 100 40 100 100 100 80 100 100 100 80 80 100 at University of California, Davis - Library on September 12, 2012 49C17A + + + + + + + + + + + + + + ranged from 0.0 to 8.0% with the associ- 49C18F + + + + + ated chi-square values of 9.3-22.5. A high 49C19E + + + chi-square value, 11.2, was observed be- 49C20D + 100 100 100 100 4OC21D 100 100 100 100 100 tween CD8A and a marker on feline chro- 49C22E + mosome B3, but the discordance was 49C23A + + + + 49C24D 70 70 50 30 60 high, 19%. 49C25A + + + + MYC, HMBS, and THRA1 had strong con- cordance and chi-square values for mark- Each cell hybrid was karyoptyped using high-resolution G-trypsIn banding and scored for the presence of every ers on feline chromosomes F2, Dl, and A3, cat chromosome. "+" Indicates ^30% of metaphases had Indicated chromosome. Numbers are actual percentages of examined metaphase spread that retained the chromosomes In the specific cell hybrid passage 17T series are respectively. These three genes also had a mouse RAG cell X feline lymphocyte hybrids. 49C series are Chinese hamster E36 X feline lymphocyte hybrids high concordancy value with one marker (O'Brien and Nash 1982). not located on the designated chromo- some, but other markers from the other film (Kodak X-OMAT XRP-5) for 2-7 days square values were calculated from a 2 x syntenic group were highly discordant. and developed using a AFP Imaging Mini/ 2 contingency table where marginal fre- For example, the concordance for THRA1 Med 90 X-ray film processor. The purified quencies were used to estimate expected to chromosome A3 and to nine other DNA samples from the hybrid lines were values. Gene symbols are as determined genes on chromosome A3 was 87-92% digested with diagnostic enzymes and the by the human nomenclature committee with corresponding chi-square values of digested products were separated, blot- (Fasman et al. 1996; McAlplne et al. 1994). 6.7-21.4. THRA1 was 88% concordant with ted, and hybridized as described above. the feline X chromosome with a chi-square Each hybrid line was scored as positive of 11.36, but the only other X marker, Results for a gene if a unique feline hybridization G6PD, was 68% discordant with a chi- pattern was seen and negative if no hy- Thirty-five genes were assigned to chro- square of 7.77. bridization pattern for the cat was detect- mosomes in the feline genome. The gene The 35 genes localized to 14 of the 19 ed. If hybridization signals were difficult to assignments were made by Southern blot feline chromosomes (Figure 2). Thirty-one score, the Southern blot was repeated us- analyses of heterologous probes on a pan- genes mapped to positions within previ- ing DNA from a different culture passage el of DNA from 45 rodent x feline somatic ously known conserved syntenic seg- of the same hybrid line. Scores were cell hybrid lines. The assignments were ments that occur between the feline and checked for concordancy and discordancy based upon concordant association of human genome. MYC is the first gene to with all other known markers typed in the each gene hybridization signal with feline be mapped to feline chromosome F2. hybrid lines including chromosomes, iso- chromosomes and markers that were pre- Three feline genes—THRA1, MOS, and zymes, and other genes (Table 2). Chi- viously mapped in the hybrid panel (Fig- PDGFB—were asyntenic with markers that

410 The Journal erf Heredity 1997:88(5) O O O CD Q Q on 2 m to r» _ — eo (O rt co co o 3 1 Kb Kb 23.2 9.4 6.7 Cat -». _ Cat- 6.7 Hamster -». ( 44 Hamster Mouse 2.3 2.0

INT1 MYB

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. 44

ARAF1 YES1 • • - • http://jhered.oxfordjournals.org/

uooomuu i 0)omvm(Dr 7T12 F 7T26 C 7T27 C 7T28 C 7T29 C 7T30 D 7T33 D 7T34 C 7T35 C 7T36 C 7T37 D :a t Mous e 0 O S S . Kb

1 •«- 6.7 Mouse -• Cat-*. 111 1 <- 4.4 | | | | | 1 Cat-* Mouse -* I m 1 <- 2.3 m mm ••- 2.0 Cat-* at University of California, Davis - Library on September 12, 2012 FGF3 1 GLI

(J O UJ UJ UJ UJ o O £ c S 106 * s

49C 1 49C 3 49C 1 49C 1 Ca t 01 O> O> 49C 4 49C 5 49C 7 49C 9 X X i Kb Hamster -» — •• — 23.1 Cat -• —•

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Hamster _». JUN - + ABL1 Figure 1. Southern blot analyses of 10 genes In the feline and rodent somatic cell hybrid panel. Gene abbreviations follow human nomenclature (McAlpine et al. 1994; Fasman et al. 1996) and are defined In Table 3.

OBrien et al • Comparative Gene Mapping in Domestic Cat 411 Table 3. Syntenic assignments of 37 genes in the feline genome

Chromosome location Discordancy Gene Discordancy range* No. of range/other symbol Gene name Human Cat Chi-square range* hybrids chromosomes' ABL1 Abelson murine leukemia viral (v-abl) oncogene homologue 1 9q34.1 D4(iy 0.05/25.4 41 0.19-0.21/11.1-8.6 ARAF1 Murine sarcoma 3611 viral (v-raf) oncogene homologue 1 Xpll.3-pll.23 X(2) 0.10-0.13/19.8-18.9 40 0.19-0.26/8.7-5.3 CD8A CD8 antigen, alpha polypeptlde (p32) 2pl2 A3(4) 0.00-8.0/22.5-9.3 37 0.19-0.27/11.2-7.3 COL1A1 Collagen, type 1, alpha 1 17q213-q22 El(l) 0.10/55 31 0.26/4.1 CSFIR Colony stimulating Factor Receptor FMS Oncogene 5q33.2q34 Al(2) 0.00-0.03/29.2-14.2 37 0.19-031/3.03-9.4 EGFR Epidermal growth factor receptor (v-erb-b) 7pl2 A2(2) 0.03-0.10/25.4-16.2 38 0.13-0.15/10.4-03 EGR1 Early growth response 1 5q23-q31 Al(2) 0.06-0.17/8.5-12.8 35 0.30-0.29/3.7-2.7 F9 Coagulation lactor K Xq26.3^27.1 X(2) 0.03-0.05/30.1-25.8 39 0.24-0.25/9.4-7 9 FES Feline sarcoma (Synder-Thellen) viral (v-fes)/Fujinaml avian sarcoma (PRCD) 15q26.1 B3<3) 0.03/32.4-30.5 40 0.11-0.21/13.5-93 viral (v-fps) oncogene homologue FCF1 FIbroblast growth factor 1 (acidic) 5q313-q33.2 A 1(2) 0.00-O.05/27.6-13.4 39 0.24-0.31/6.6-35 FGF3 FIbroblast growth factor 2, murine mammary tumor Integration site (v-lnt-2) 1 Iql33 Dl(3) 0.00-0.06/28.2-11.0 35 0.15-0.20/14.1^1.9 oncogene homologue FOS FBJ murine osteosarcoma viral (v-fos) oncogene homologue 14q24.3 B3(3) 0 08-0.05/233-26.8 38 0.22-0.26/8.6-2.4

fflV FYN oncogene related to SRC, FGR, YES, SYR 6q21 B2(4) 0.05-0.10/275-14.2 40 0.20-0.26/8.6-2.4 Downloaded from GU Glloma-assoclated oncogene homologue 12ql3 B4(4) 0.08-0.12/24.6-7.6 41 0.29-0.35/43-0.4 H0XA4 HomeoboxA4 7pl5-pl4 A2(l) 0.06/16.3 36 0.11/2.4 HRAS Harvey rat sarcoma viral (v-Ha-ras) oncogene homologue Ilpl5.5 Dl(4) 0.10-0.19/8.9-6.0 21 0.23-0.29/5.2-3.7 HRASP Harvey rat sarcoma viral (v-Ha-ras) oncogene homologue pseudogene Xpl 1.3-pl 1^3 X(2) 0.07/8.1-7.3 15 0.14-0.29/5.0-1.0 ILIA lnterleukln 1, alpha 2ql3 A3(4) 0.00/34.5-10.5 40 0.15-0.20/15.5-2.9 US lnterleukln 8 4ql3-q21 Bl(2) 0.11-0.12/17.0-16.0 36 0.22-0.43/6.9-2.0 JUN Avian sarcoma virus 17 (v-Jun) oncogene homologue Ip32-p31 Cl(4) 0.06-0.11/20.1-14.1 35 0.11-0.70/5.4-0.1 KIT Hardy-Zuckerman 4 feline sarcoma viral (v-klt) oncogene homologue 4ql2 Bl(2) 0.05-0.16/14.0-12.7 37 0.18-0.33/9.7-1.3 http://jhered.oxfordjournals.org/ KRAS2 Klrsten rat sarcoma 2 viral homologue (v-KI-ras2) oncogene 12pl2.1 B4(3) 0.00-0.09/8.7-3.9 13 0.15-0.29/3.3-0 1 MOS Moloney leukemia sarcoma virus (v-mos) oncogene homologue 8qll B2(4) 0.03-0.14/29.6-12.9 38 0.16-0.26/11.6-4.7 MYB Avian myelobastosls viral (v-myb) oncogene homologue 6q23.3-q24 B2(4) 0.06-0.17/23.8-10.8 39 0.23-0.32/6.7-2.3 MYC Avian myelocytomatosis viral (v-myc) oncogene homolog 8q24.12-q24.13 F2(l) 0.03/26.3 35 0.00-0.41/10.2-2.6 MYCN Avian myelocytomatosis viral related oncogene 2p24.3 A3(4) 0 05/25 4-11.7 39 0.21-0.28/11.8-9.7 NRAS Neuroblastoma Ras viral (v-ras) oncogene homologue Ipl3 Cl(4) 0.05-0.09/24.1-14.9 40 0.12-0.62/115-0.2 OTC Ornlthlne transcarbomoyi-transferase Xp21.1 X(2) 0.10-0.15/215-16.8 41 0.27-0.36/8.9-3.7 PDGFB Platelet-derived growth factor beta polypeptlde (sis oncogene) 22ql2.3-ql3.1 B4(4) 0.03-0.05/30.4-11.0 38 026-0.34/5.9-0.8 PIM1 Plm-1 oncogene 6p21 B2(4) 0.06-0.15/24.6-11.8 39 0.18-0.53/ ROS1 Avian UR2 sarcoma virus oncogene (v-ros) homologue 6q21-q22 B2(4) 0.00-0 16/25.3-10.1 23 0.18-0.21/8.1-7.4 SRC Avian sarcoma (Schmldt-Ruppln A-2) viral (v-src) oncogene homologue 2Oqll.2 A3(4) 0.05-0.07/28.2-10.9 41 0.13-0.20/18S-13.0 THRA1 Thyroid hormone receptor, alpha 1 (avian erythroblastlc leukemia viral • 17qll.2-o.12 A3(9) 0.08-0.13/6.7-21.4 37 0.12-0.68/11.4-7.8 (v-erb-a) oncogene homologue 1, (ERBA1) at University of California, Davis - Library on September 12, 2012 WNT1 Murine mammary tumor Integration site (vMnt-1) oncogene homologue 12ql3 B4(4) 0.00-0.06/29.9-10.1 34 0.25-0.47/6.9-0.1 YES1 Yamaguchl sarcoma viral (v-yes-1) oncogene homologue 18pll.31-pll.22 D3(l) 0.03/28.2 38 0.22-0.28/9.7-1.8

• Discordancy range: Range of measured frequency of hybrids' discordance for tested gene versus Implicated chromosome by G-banded karyology plus test gene vs. other gene markers previously mapped to that chromosome. kChl square: Test for random occurrence of hybrids In four categories with respect to gene/marker combinations: +/+, +/-, — /+, and — /—. ' Discordancy range: Range of hybrids discordant for gene versus all other cat chromosomes. ' In paretheses Is number of gene markers previously mapped to implicated chromosome that were typed for concordance with the mapped gene

are syntenic in humans. THRA1, a gene on chromosome D4. The 35 assignments in- T, Cevario S, O'Brien SJ, Tsujimoto H, and human chromosome 17, mapped to A3. crease the syntenic map of the cat to 104 Hasegawa A, submitted; Yuhki and O'Brien Two other human chromosome 17 mark- loci, representing 18 of the 19 feline chro- 1988), 2 homeobox genes (Masuda et al. ers—TP53 and COL1A1—mapped to feline mosomes. Ninety-six of the genes mapped 1991), 1 gene encoding an rRNA (Yu et al. chromosome El. Feline chromosome A3 in the cat have a human homologue. Only 1980), 2 genes encoding coat-color phe- now has markers from three different hu- two small chromosomes, Fl and E2, do not notypes (O'Brien et al. 1986), 4 copies of man chromosomes—17, 20, and 2. MOS have genes localized to them. the endogenous retrovlrus RD114 (Reeves mapped to feline chromosome B2 in con- et al. 1985), and 13 miscellaneous genes. trast to the other human chromosome 8 This increase in markers on the feline map Discussion markers, GSR and MYC, which are located aids disease gene analyses, the study of on feline chromosomes C2 and F2, respec- We have added 35 genes to the feline ge- functional genome organization, and the tively. Feline chromosome B2 has an ex- netic map by Southern blot analyses of a extent of possible genome comparisons. tensive human chromosome 6 gene clus- feline x rodent somatic cell hybrid panel. The 35 genes localized to 14 different fe- ter and MOS is the only marker on B2 that These assignments increase the marker line chromosomes. Each feline chromo- is from a different human chromosome. density of the syntenic map of the cat to some, except for E2 and Fl, now has at PDGFB is the only marker on feline chro- 105 loci; 36 isozymes (Berman et al. 1986; least one genetic marker with a homolo- mosome B4 that is not from human chro- Gilbert et al. 1988; O'Brien and Nash 1982), gous gene mapped in humans. Only hu- mosome 12. YES] from human chromo- 35 oncogenes (Okuda et al. 1993; Tsuji- man chromosomes 16 and 19 are not rep- some 18 and ABL1 from human chromo- moto et al. 1993), 12 genes involved with resented in the feline map. Human chro- some 9 are the only markers mapped in immune response (Cho KW, Youn HY, Cev- mosomes 9, 13, and 18 are represented in the cat from these human chromosomes. ario S, O'Brien SJ, Watari T, Tsujimoto H, the cat genome by one marker each. Hu- ABL1 represents the third human chro- and Hasegawa A, submitted; Okuda M, Mi- mans have 24 chromosomes, including Y, mosome with gene homologues on feline nehata K, Setoguchl A, Nishigaki K, Watari thus 24 different syntenic groups. A direct

412 The Journal of Heredity 1997.88(5) tended from the short arm to the long arm ESD [13c * THRA1 17g Boxed rrPA 20p of the human chromosome. For example, RDV2 - RAF1 *M SRC 20q Human chromosome B4 has four human 12p ARSB 5p W ADA 2Qq Chromosomes genes and four human 12q genes. Conser- HOXA.4 « ACPI 2p EGR1 TCRG *R MYCN vation of synteny across the human cen- FGF1 EGFR B MDH1 2p tromere is also reflected by feline chro- CSF1R MET ~ REL 2p TCRB *AQ CD8A 2p mosomes Al, A2, A3, Bl, B2, B4, Dl, El, "° IGKC 2p andX. * IL1A Ninety-one of the 105 genes mapped in FTT1 _ the cat are also localized In the mouse and 9 MOS • 4p MANA1 15q CDKN1B 12p human genomes. These 91 genes are PEPS MHCO 6p KIT 4q HEXA 15q _ GAPO 12p •— CDKN1A 6p found on 21 different human chromo- IL8 4q MPI 15q TPM 12p PUU11 6p somes, 18 different feline chromosomes, IL2 4q PKM2 15q £\ LDHB 12p GLO1 6p IDH2 15q KRAS2 Ii2p m ME1 6q *\\ and 19 of 21 mouse chromosomes. Feline FES 15g LJ HOXC.1 12q chromosome A3 genes disperse to three PGM3 6q NP 14q WNT1 I2q

B1 Downloaded from ROS1 6q „ TCRA 14q * ^ GU 42q human chromosomes and to four different MYB 6q B3 TCRD 14q B4 PEPB H2q FYN 6q mouse chromosomes (2, 6, 11, and 12). Fe- SOD2 * FOS 14q * PDGFB PGM1 1p 6q IGH© 14q line chromosome B2 has genes from two m PGD 1P m GLB1 3q human chromosomes and these genes are • FUCA1 1P SOD1 21q JUN 1p RDV1 ETS2 21q RDV3 found on four different mouse chromo- • NRAS 1p somes (4, 9, 10, and 17). Markers on feline RDV4 http://jhered.oxfordjournals.org/ chromosome B4 are found on three differ- IDH1 "2q] GSR ent mouse chromosomes. All mouse and C1 human X-linked gene homologues were Ir HRAS mapped to the feline X chromosome. The HBB conserved localization of genes on the X LDHA ACP2 HK1 |iOg 12a chromosome by over 19 eutherian mam- *l—I FGF3 4p mal species is considered a selective TYR NUMT - 9q HMBS mechanism to compensate for X-chromo- D1 ETS1 some inactivation (DeBry and Seldin 1996; GANAB Mouse Genome Database). at University of California, Davis - Library on September 12, 2012 Seventy-four feline genes are Included in rRNA blocks of two or more markers from the TP53 17p GUSB LJJ1J * OTC Xp same human chromosome representing 15 COL1A1 17q *^ ARAF1 Xp *U HRASP Xp conserved syntenic segments between hu- X GLA Xq man and cat genomes. The same 74 gene LJ HPRT Xq homologues are found on 17 mouse chro- MYC SRY Y *^n F9 Xq ZFY Y H G6PD Xq mosomes; but the segments are often sepa- ^ BVRI X 0 - rated by genes from a second human chro- mosome, disrupting conserved blocks Figure 2. Syntenlc map of the domestic cat (Felis calus). Ideograms depict the 19 feline chromosomes (2N = 38). Gene assignments do not convey distance, order, or regional localization on the feline chromosomes, but seen in the cat to 30 conserved segments parallel the order found In humans, as determined cytogenetlcally. Human localizations are boxed to the right of between mouse and human (Copeland et gene symbols. Asterisks Indicate genes mapped In this study. Gene symbols reflect human nomenclature (McAlplne al. 1993; DeBry and Seldin 1996). These et al. 1994, Fasman et al. 1996). data suggest that the cat is two to three times less genomically rearranged than correspondence of these 24 syntenic regions of the human chromosomes. The the mouse when compared to humans. groups to the 20 chromosomes (including overall conservation of syntenic groups The addition of markers to the feline ge- Y) in the feline genome would require four between humans and cats is strong and netic map and the strong genomic conser- feline chromosomes to represent at least most of the asyntenic genes are from vation to the human genome makes the two human syntenic groups each. Our regions with high potential for rearrange- cat a valuable animal model for studying data shows that 11 feline chromosomes ment. inherited diseases. The comparative gene are represented by markers from at least An exception is observed with human mapping approach for disease studies is a two different human chromosomes. Only chromosome 8 syntenic groups. Three more efficient method for candidate gene 2 of these 11 feline chromosomes, A3 and chromosome 8 markers have been identification than genome-wide linkage B3, are represented by more than one mapped in the cat and each localizes to a studies, particularly in species with small marker from two different human syntenic different feline chromosome. The three mapping projects. The cat is a model for groups. A majority of the feline chromo- chromosome 8 markers—GSR, MOS, and over 30 inherited diseases found In hu- somes that have markers from more than MYC—are from three different regions of mans (Nicholas et al. 1996; Migaki et al. one human chromosome have only one human chromosome 8—8p, 8cen, and 1992). Many of these feline disease models marker representing the second syntenic 8q—which may explain the disruption. complement murine models, and several group. Most of these isolated markers are But most of the larger blocks of conserved disease models are unique to cat and hu- from extremely telomeric or centromeric synteny between cats and humans has ex- mans. The current genetic map of the cat

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