- Email: [email protected]
The Human CC Chemokine TECK (SCYA25) Maps to Chromosome 19p13.2
311
BRIEF MAPPING REPORTS
proliferation (4). In the present study, the human PTPRK gene was assigned to 6q22.2– q22.3, a region frequently deleted in hematological neoplasms, melanomas, ovary carcinomas, and many other solid tumors. As regards this chromosomal location and the biochemical properties of its product, the human PTPRK gene might be a good candidate for the presumed tumor suppressor gene in the long arm of chromosome 6. References 1.
2.
3.
4.
5.
Harder, J., Siebert, R., Zhang, Y., Matthiesen, P., Christophers, E., Schlegelberger, B., and Schro¨der, J-M. (1997). Mapping of the gene encoding human b-defensin-2 (DEFB2) to chromosome region 8p22–p23.1. Genomics, 46: 472– 475. Sahar, E., and Latt, S. A. (1980). Energy transfer and binding competition between dyes used to enhance staining differentiation in metaphase chromosomes. Chromosoma 79: 1–28. Sorio, C., Melotti, P., D’Arcangelo, D., Mendrola, J., Calabretta, B., Croce, C. M., and Huebner, K. (1997). Receptor protein tyrosine phosphatase gamma, PTPr, regulates hematopoietic differentiation. Blood 90: 49 –57. Yang, Y., Gil, M., Byun, S. M., Choi, I., Pyun, K. H., and Ha, H. (1996). Transforming growth factor-b1 inhibits human keratinocyte proliferation by upregulation of a receptor-type tyrosine phosphatase R-PTP-k gene expression. Biochem. Biophys. Res. Commun. 228: 807– 812. Yang, Y., Gil, M. C., Choi, E. Y., Park, S. H., Pyun, K. H., and Ha, H. (1997). Molecular cloning and chromosomal localization of a human gene homologous to the murine R-PTP-k, a receptor-type protein tyrosine phosphatase. Gene 186: 77– 82.
The Human CC Chemokine TECK (SCYA25) Maps to Chromosome 19p13.2 H. Nomiyama,*,1 K. Amano,† J. Kusuda,‡ T. Imai,§ R. Miura,* O. Yoshie,§,2 and Y. Matsuda† *Department of Biochemistry, Kumamoto University Medical School, 2-2-1 Honjo, Kumamoto 860-0811 Japan; †Laboratory of Animal Genetics, School of Agricultural Sciences, Chikusa, Nagoya 464-8601, Japan; ‡Laboratory of Genetic Resources, National Institute of Health, 1-23-1 Toyama, Shinjuku-ku, Tokyo 162-8640, Japan; and §Shionogi Institute for Medical Science, 2-5-1 Mishima, Settsu-shi, Osaka 566-0022, Japan Received April 1, 1998; accepted April 16, 1998
Functional gene description: Chemokines are a large family of cytokines that regulate leukocyte trafficking 1 To whom correspondence should be addressed. Telephone: 81-96373-5063. Fax: 81-96-372-6140. E-mail: [email protected]. 2 Current address: Department of Microbiology, Kinki University School of Medicine, 377-2 Ohno-higashi, Osaka-Sayama, Osaka 5898511, Japan.
GENOMICS 51, 311–312 (1998) ARTICLE NO. GE985352
0888-7543/98 $25.00 Copyright © 1998 by Academic Press All rights of reproduction in any form reserved.
(1). According to the positions of the first two of the four conserved cysteine residues, chemokines are classified into two major subfamilies, CXC and CC. Most of the human CC chemokine genes cluster at chromosome 17q11.2. CC chemokine TECK (thymus-expressed chemokine) (5) is distantly related to other CC chemokines and is expressed mainly in thymic dendritic cells. Mouse TECK has been shown to be chemotactic for activated macrophages, dendritic cells, and thymocytes. Although the mouse TECK gene has been localized to the proximal part of chromosome 8 (5), the human counterpart has not yet been mapped. DNA source: The human BAC library (Research Genetics, Huntsville, AL) with an average insert size of 130 kb was screened by PCR. The primers used were 59-GATAAAACCGTCGCCCTACA and 59-GCGGCAAAAGGTTAAAATCA (product size, 163 bp). One clone, 133j9, was obtained. Methods of mapping: For SCH, the NIGMS human 3 rodent somatic cell mapping panel No. 2 version 2 and PCR were used. For RH, the GeneBridge 4 panel (Research Genetics) and PCR were used (LOD . 3.0). For FISH, direct R-banding was performed using the BAC 133j9 DNA as probe. One hundred cells were examined, and 87 cells showed doublet signals on homologous chromosomes. Results: By SCH and RH, the TECK gene was localized on the short arm of chromosome 19, 3.5 cR centromeric of marker RPI S28I 1 and 20.4 cR telomeric of D19S221 (not shown). D19S221 maps to a site close to chromosome 19p13.2 within p13.1 (LLNL maps of human chromosome 19 are available at http://www-bio.llnl.gov/bbrp/genome/ genome.html). A region of 19p13.2 close to p13.3 is syntenic to the mouse chromosome 8 (3). To confirm the localization, FISH was performed. The isolated BAC clone 133j9 was used as probe. An example is shown in Fig. 1. The result clearly shows that the TECK gene is located on chromosome 19p13.2. Additional comments: Recently, some CC chemokines have been mapped to chromosomes other than chromosome 17: a minicluster of TARC (SCYA17), MDC (SCYA22), and fractalkine/neurotactin (CX3C chemokine, SCYD1) at 16q13, LARC/MIP-3a/exodus-1 (SCYA20) at 2q33– q37, ELC/MIP-3 b /exodus-3 (SCYA19) and SLC/6Ckine/exodus-2 (SCYA21) at 9p13, and MPIF-2/ eotaxin-2 (SCYA24) at 7q11.23. These observations and the result described here indicate that some CC chemokine genes were scattered on several chromosomes during the evolutionary diversification and amplification of the gene family. References 1.
Baggiolini, M., Dewald, B., and Moser, B. (1997). Human chemokines: An update. Annu. Rev. Immunol. 15: 675–705.
312
BRIEF MAPPING REPORTS
FIG. 1. Chromosomal localization of the human TECK gene on chromosome 19p13.2 by FISH. The hybridization signals are indicated by arrows. R-band and G-band patterns are shown in (a, c) and (b), respectively. Chromosomes were identified by their size and G-banding patterns. Preparation of R-banded chromosomes and FISH were essentially as described (2, 4). Briefly, the entire BAC 133j9 DNA was labeled with biotin– dUTP by nick-translation. The labeled probe was mixed with excess human Cot1 DNA (Life Technologies, Rockville, MD) and hybridized to metaphase chromosomes in a solution containing 50% formamide, 23 SSC, 10% dextransulfate, and 2 mg/ml BSA at 37°C overnight. After being washed, the samples were incubated with FluoroLink Cy2 streptavidin (Amersham, Cleveland, OH) and stained with propidium iodide. 2.
Matsuda, Y., and Chapman, V. M. (1995). Application of fluorescence in situ hybridization in genome analysis of the mouse. Electrophoresis 16: 261–272.
3.
Mohrenweiser, H., Olsen, A., Archibald, A., Beattie, C., Burmeister, M., Lamerdin, J., Lennon, G., Stewart, E., Stubbs, L., Weber, J. L., and Johnson, K. (1996). Report of the Third International Workshop on Human Chromosome 19 Mapping 1996. Cytogenet. Cell Genet. 74: 161–186.
4.
Takahashi, E., Hori, T., O’Connell, P., Leppert, M., and
White, R. (1990). R-banding and nonisotopic in situ hybridization: Precise localization of the human type II collagen gene (COL2A1). Hum. Genet. 86: 14 –16. 5. Vicari, A. P., Figueroa, D. J., Hedrick, J. A., Foster, J. S., Singh, K. P., Menon, S., Copeland, N. G., Gilbert, D. J., Jenkins, N. A., Bacon, K. B., and Zlotnik, A. (1997). TECK: A novel CC chemokine specifically expressed by thymic dendritic cells and potentially involved in T cell development. Immunity 7: 291–301.
BRIEF MAPPING REPORTS
proliferation (4). In the present study, the human PTPRK gene was assigned to 6q22.2– q22.3, a region frequently deleted in hematological neoplasms, melanomas, ovary carcinomas, and many other solid tumors. As regards this chromosomal location and the biochemical properties of its product, the human PTPRK gene might be a good candidate for the presumed tumor suppressor gene in the long arm of chromosome 6. References 1.
2.
3.
4.
5.
Harder, J., Siebert, R., Zhang, Y., Matthiesen, P., Christophers, E., Schlegelberger, B., and Schro¨der, J-M. (1997). Mapping of the gene encoding human b-defensin-2 (DEFB2) to chromosome region 8p22–p23.1. Genomics, 46: 472– 475. Sahar, E., and Latt, S. A. (1980). Energy transfer and binding competition between dyes used to enhance staining differentiation in metaphase chromosomes. Chromosoma 79: 1–28. Sorio, C., Melotti, P., D’Arcangelo, D., Mendrola, J., Calabretta, B., Croce, C. M., and Huebner, K. (1997). Receptor protein tyrosine phosphatase gamma, PTPr, regulates hematopoietic differentiation. Blood 90: 49 –57. Yang, Y., Gil, M., Byun, S. M., Choi, I., Pyun, K. H., and Ha, H. (1996). Transforming growth factor-b1 inhibits human keratinocyte proliferation by upregulation of a receptor-type tyrosine phosphatase R-PTP-k gene expression. Biochem. Biophys. Res. Commun. 228: 807– 812. Yang, Y., Gil, M. C., Choi, E. Y., Park, S. H., Pyun, K. H., and Ha, H. (1997). Molecular cloning and chromosomal localization of a human gene homologous to the murine R-PTP-k, a receptor-type protein tyrosine phosphatase. Gene 186: 77– 82.
The Human CC Chemokine TECK (SCYA25) Maps to Chromosome 19p13.2 H. Nomiyama,*,1 K. Amano,† J. Kusuda,‡ T. Imai,§ R. Miura,* O. Yoshie,§,2 and Y. Matsuda† *Department of Biochemistry, Kumamoto University Medical School, 2-2-1 Honjo, Kumamoto 860-0811 Japan; †Laboratory of Animal Genetics, School of Agricultural Sciences, Chikusa, Nagoya 464-8601, Japan; ‡Laboratory of Genetic Resources, National Institute of Health, 1-23-1 Toyama, Shinjuku-ku, Tokyo 162-8640, Japan; and §Shionogi Institute for Medical Science, 2-5-1 Mishima, Settsu-shi, Osaka 566-0022, Japan Received April 1, 1998; accepted April 16, 1998
Functional gene description: Chemokines are a large family of cytokines that regulate leukocyte trafficking 1 To whom correspondence should be addressed. Telephone: 81-96373-5063. Fax: 81-96-372-6140. E-mail: [email protected]. 2 Current address: Department of Microbiology, Kinki University School of Medicine, 377-2 Ohno-higashi, Osaka-Sayama, Osaka 5898511, Japan.
GENOMICS 51, 311–312 (1998) ARTICLE NO. GE985352
0888-7543/98 $25.00 Copyright © 1998 by Academic Press All rights of reproduction in any form reserved.
(1). According to the positions of the first two of the four conserved cysteine residues, chemokines are classified into two major subfamilies, CXC and CC. Most of the human CC chemokine genes cluster at chromosome 17q11.2. CC chemokine TECK (thymus-expressed chemokine) (5) is distantly related to other CC chemokines and is expressed mainly in thymic dendritic cells. Mouse TECK has been shown to be chemotactic for activated macrophages, dendritic cells, and thymocytes. Although the mouse TECK gene has been localized to the proximal part of chromosome 8 (5), the human counterpart has not yet been mapped. DNA source: The human BAC library (Research Genetics, Huntsville, AL) with an average insert size of 130 kb was screened by PCR. The primers used were 59-GATAAAACCGTCGCCCTACA and 59-GCGGCAAAAGGTTAAAATCA (product size, 163 bp). One clone, 133j9, was obtained. Methods of mapping: For SCH, the NIGMS human 3 rodent somatic cell mapping panel No. 2 version 2 and PCR were used. For RH, the GeneBridge 4 panel (Research Genetics) and PCR were used (LOD . 3.0). For FISH, direct R-banding was performed using the BAC 133j9 DNA as probe. One hundred cells were examined, and 87 cells showed doublet signals on homologous chromosomes. Results: By SCH and RH, the TECK gene was localized on the short arm of chromosome 19, 3.5 cR centromeric of marker RPI S28I 1 and 20.4 cR telomeric of D19S221 (not shown). D19S221 maps to a site close to chromosome 19p13.2 within p13.1 (LLNL maps of human chromosome 19 are available at http://www-bio.llnl.gov/bbrp/genome/ genome.html). A region of 19p13.2 close to p13.3 is syntenic to the mouse chromosome 8 (3). To confirm the localization, FISH was performed. The isolated BAC clone 133j9 was used as probe. An example is shown in Fig. 1. The result clearly shows that the TECK gene is located on chromosome 19p13.2. Additional comments: Recently, some CC chemokines have been mapped to chromosomes other than chromosome 17: a minicluster of TARC (SCYA17), MDC (SCYA22), and fractalkine/neurotactin (CX3C chemokine, SCYD1) at 16q13, LARC/MIP-3a/exodus-1 (SCYA20) at 2q33– q37, ELC/MIP-3 b /exodus-3 (SCYA19) and SLC/6Ckine/exodus-2 (SCYA21) at 9p13, and MPIF-2/ eotaxin-2 (SCYA24) at 7q11.23. These observations and the result described here indicate that some CC chemokine genes were scattered on several chromosomes during the evolutionary diversification and amplification of the gene family. References 1.
Baggiolini, M., Dewald, B., and Moser, B. (1997). Human chemokines: An update. Annu. Rev. Immunol. 15: 675–705.
312
BRIEF MAPPING REPORTS
FIG. 1. Chromosomal localization of the human TECK gene on chromosome 19p13.2 by FISH. The hybridization signals are indicated by arrows. R-band and G-band patterns are shown in (a, c) and (b), respectively. Chromosomes were identified by their size and G-banding patterns. Preparation of R-banded chromosomes and FISH were essentially as described (2, 4). Briefly, the entire BAC 133j9 DNA was labeled with biotin– dUTP by nick-translation. The labeled probe was mixed with excess human Cot1 DNA (Life Technologies, Rockville, MD) and hybridized to metaphase chromosomes in a solution containing 50% formamide, 23 SSC, 10% dextransulfate, and 2 mg/ml BSA at 37°C overnight. After being washed, the samples were incubated with FluoroLink Cy2 streptavidin (Amersham, Cleveland, OH) and stained with propidium iodide. 2.
Matsuda, Y., and Chapman, V. M. (1995). Application of fluorescence in situ hybridization in genome analysis of the mouse. Electrophoresis 16: 261–272.
3.
Mohrenweiser, H., Olsen, A., Archibald, A., Beattie, C., Burmeister, M., Lamerdin, J., Lennon, G., Stewart, E., Stubbs, L., Weber, J. L., and Johnson, K. (1996). Report of the Third International Workshop on Human Chromosome 19 Mapping 1996. Cytogenet. Cell Genet. 74: 161–186.
4.
Takahashi, E., Hori, T., O’Connell, P., Leppert, M., and
White, R. (1990). R-banding and nonisotopic in situ hybridization: Precise localization of the human type II collagen gene (COL2A1). Hum. Genet. 86: 14 –16. 5. Vicari, A. P., Figueroa, D. J., Hedrick, J. A., Foster, J. S., Singh, K. P., Menon, S., Copeland, N. G., Gilbert, D. J., Jenkins, N. A., Bacon, K. B., and Zlotnik, A. (1997). TECK: A novel CC chemokine specifically expressed by thymic dendritic cells and potentially involved in T cell development. Immunity 7: 291–301.