Skip to main content
Log in

Cloning of the β2-microglobulin gene in the zebrafish

  • Original Articles
  • Published:
Immunogenetics Aims and scope Submit manuscript

Abstract

The β2-microglobulin (β2m) is a protein found in the serum in a free form and on the cell surface in a form noncovalently associated with the α chain of the class I major histocompatibility complex (Mhc) molecules. In mammals, the β2m-encoding gene (B2m) is found on a chromosome different from the Mhc proper. We have isolated and characterized the B2m gene of the zebrafish, Brachydanio rerio, family Cyprinidae. We obtained both cDNA and genomic clones of the Brre-B2m gene. The cDNA clones contained the entire coding sequence, the entire 3′ untranslated (UT) region, and at least part of the 5′UT region. The genomic clone contained the entire Brre-B2m gene. The coding sequence specifies 97 amino acid residues of the mature protein so that the zebrafish β2m is two residues shorter than human and one residue shorter than cattle, fowl, or turkey β2m (codons at positions 85 and 86 have been deleted in the Brre-B2m. gene). The amino acid and nucleotide sequence similarities between zebrafish and human β2m (B2m) are 45% and 59%, respectively. Approximately 24% of the positions are invariant and an additional 9% show only conservative substitutions in comparisons which include all known β2m sequences (fish, avian, and mammalian). Most of the conserved positions are in the β strands (some 47% of the β-strand positions are conserved in the three vertebrate classes). The Brre-B2m gene consists of four exons separated by three introns. All of the introns are considerably shorter than the corresponding introns in the mammalian B2m genes. The coding sequences of the cDNA and the genomic clones are almost identical but the sequences of the 3'UT regions differ at 1.7% of the sites, suggesting that the genes borne by these clones might have diverged at least 0.7 million years (my) ago. In contrast to the human B2m gene, the Brre-B2m gene shows no bias in the distribution of the CpG dinucleotides: the dinucleotides are distributed evenly along the entire available sequence. The haploid genome of the zebrafish contains only one copy of the B2m gene.

This is a preview of subscription content, log in via an institution to check access.

Access this article

Price excludes VAT (USA)
Tax calculation will be finalised during checkout.

Instant access to the full article PDF.

Similar content being viewed by others

References

  • Arce-Gomez, B., Jones, E. A., Barnstable, C. J., Solomon, E., and Bodmer, W. F. The genetic control of HLA-A and B antigens in somatic cell hybrids: Requirement for β2. Tissue Antigens 11: 96–112, 1978

    Google Scholar 

  • Becker, J. W. and Reeke, G. N. Three-dimensional structure of β2-microglobulin. Proc Natl Acad Sci USA 82: 4225–4229, 1985

    Google Scholar 

  • Berggård, I. and Bearn, A. G. Isolation and properties of a low molecular weight β2-globulin occurring in human biological fluids. J Biol Chem 243: 4095–4103, 1968

    Google Scholar 

  • Bjorkman, P. J., Saper, M. A., Samraoui, B., Bennett, W. S., Strominger, J. L., and Wiley, D. C. Structure of the human class I histocompatibility antigen, HLA-A2. Nature 329: 506–512, 1987

    Google Scholar 

  • Brinckerhoff, C. E., Mitchell, T. I., Karmilowicz, M. J., Kluve-Beckerman, B., Benson, M. D. Autocrine induction of collagenase by serum amyloid A-like and β2-microglobulin-like proteins. Science 243: 655–657, 1989

    Google Scholar 

  • Burnet, F. M. A certain symmetry: histocompatibility antigens compared with immunocyte receptors. Nature 226: 123–126, 1970

    Google Scholar 

  • Calabi, F. and Milstein, C. A novel family of human major histocompatibility complex-related genes not mapping to chromosome 6. Nature 323: 540–543, 1986

    Google Scholar 

  • Cejka, J., Van Nieuwkoop, J., Mood, D., Kithier, K., and Radl, J. β2-Microglobulin in human colostrum and milk: effect of breast feeding and physiochemical characterization. Clin Chim Acta 67: 71–78, 1976

    Google Scholar 

  • Cunningham, B. A., Wang, J. L., Berggård, I., and Peterson, P. A. The complete amino acid sequence of β2-microglobulin. Biochemistry 12: 4811–4822, 1973

    Google Scholar 

  • Daniel, F., Morello, D., Le Bail, O., Chambon, P., Cayre, Y., and Kourilsky, P. Structure and expression of the mouse β2-microglobulin gene isolated from somatic and non-expressing teratocarcinoma cells. EMBO J 2: 1061–1065, 1983

    Google Scholar 

  • Dargemont, C., Dunon, D., Deugnier, M.-A., Denoyelle, M., Girault, J.-M., Lederer, F., Lê, K. H. D., Godeau, F., Thiery, J. P., and Imhof, B. A. Thymotaxin, a chemotactic protein, is identical to β2-microglobulin. Science 246: 803–806, 1989

    Google Scholar 

  • Dixon, B., Stet, R. J. M., van Erp, S. H. M., and Pohajdak, W. Characterization of β2-microglobulin transcripts from two teleost species. Immunogenetics 38: 27–34, 1993

    Google Scholar 

  • Dobberstein, B., Garoff, H., and Warren, G. Cell-free synthesis and membrane insertion of mouse H-2Dd histocompatibility antigen and β2-microglobulin. Cell 17: 759–769, 1979

    Google Scholar 

  • Dunon, D., Kaufman, J., Salomonsen, J., Skjødt, K., Vainio, O., Thiery, J.-P., and Imhof, B. A. T cell precursor migration towards β2-microglobulin is involved in thymus colonization of chicken embryos. EMBO J 9: 3315–3322, 1990

    Google Scholar 

  • Feinberg, A. P. and Vogelstein, B. A technique for radiolabeling DNA restriction endonuclease fragments to high specific activity. Anal Biochem 132: 6–13, 1983

    CAS  PubMed  Google Scholar 

  • Gally, J. A. and Edelman, G. M. The genetic control of immunoglobulin synthesis. Annu Rev Genet 6: 1–46, 1972

    Google Scholar 

  • Gates, F. T. III, Coligan, J. E., and Kindt, T. J. Complete amino acid sequence of rabbit β2-microglobulin. Biochemistry 18: 2267–2272, 1979

    Google Scholar 

  • Gold, J. R. and Karel, J. DNA base composition and nucleotide distribution among fifteen species of teleostean fishes. Comp Biochem Physiol 90B: 715–719, 1988

    Google Scholar 

  • Goodfellow, P. N., Jones, E. A., van Heyningen, V., Solomon, E., Bobrow, M., Miggiano, V., and Bodmer, W. F. The β2-microglobulin gene is on chromosome 15 and not in the HL-A region. Nature 254: 267–269, 1975

    Google Scholar 

  • Grey, H. M., Kubo, R. T., Colon, S. M., Poulik, M. D., Cresswell, P., Springer, T., Turner, M., and Strominger, J. L. The small subunit of HL-A antigens is β2 microglobulin. J Exp Med 138: 1608–1612, 1973

    Google Scholar 

  • Groves, M. L. and Greenberg, R. Bovine homologue of β2-microglobulin isolated from milk. Biochem Biophys Res Comm 77: 320–327, 1977

    Google Scholar 

  • Groves, M. L. and Greenberg, R. Complete amino acid sequence of bovine β2-microglobulin. J Biol Chem 257: 2619–2626, 1982

    Google Scholar 

  • Groves, M. L., Basch, J. J., and Gordon, W. G. Isolation, characterization, and amino acid composition of a new crystalline protein, lactollin, from milk. Biochemistry 2: 814–817, 1963

    Google Scholar 

  • Güssow, D., Rein, R., Ginjaar, I., Hochstenbach, F., Seemann, G., Kottman, A., and Ploegh, H. L. The human β2-microglobulin gene: Primary structure and definition of the transcriptional unit. J Immunol 139: 3132–3138, 1987

    Google Scholar 

  • Hashimoto, K., Nakanishi, T., and Kurosawa, Y. Isolation of carp genes encoding major histocompatibility complex antigens. Proc Natl Acad Sci USA 87: 6863–6867, 1990

    Google Scholar 

  • Holmes, D. S. and Quigley, M. A rapid boiling method for the preparation of bacterial plasmids. Anal Biochem 114: 193–197, 1981

    Google Scholar 

  • Kaufman, J., Andersen, R., Avila, D., Engberg, J., Lambris, J., Salomonsen, J., Welinder, K., and Skjødt, K. Different features of the Mhc class I heterodimer have evolved at different rates. J Immunol 148: 1532–1546, 1992

    CAS  PubMed  Google Scholar 

  • Klein, C., Ono, H., Goldschmidt, T., O'hUigin, C., Vincek, V., and Klein, J. Extensive Mhc variability in cichlid fishes of Lake Malawi. Nature, submitted, 1992

  • Klein, J., Ono, H., Klein, D., and O'hUigin, C. The accordion model of Mhc evolution. In J. Klein and D. Klein (eds.): Progress in Immunology vol. 8, Springer. Heidelberg, in press, 1992

    Google Scholar 

  • Kyte, J. and Doolittle, R. F. A simple method for displaying the hydropathic character of a protein. J Mol Biol 157: 105–132, 1982

    CAS  PubMed  Google Scholar 

  • Lawlor, D. A., Warren, E., Ward, F. E., and Parham, P. Comparison of class I MHC alleles in humans and apes. Immunol Rev 113: 147–185, 1990

    Google Scholar 

  • Li, W.-H., Wu, C.-I., and Luo, C.-C. A new method for estimating synonymous and nonsynonymous rates of nucleotide substitution considering the relative likelihood of nucleotide and codon changes. Mol Biol Evol 2: 150–174, 1985

    Google Scholar 

  • Lingappa, V. R., Cunningham, B. A., Jazwinski, M., Hopp, T. P., Blobel, G., and Edelman, G. M. Cell-free synthesis and segregation of β2-microglobulin. Proc Natl Acad Sci USA 76: 3651–3655, 1979

    Google Scholar 

  • Lögdberg, L., Björck, J., and Cigen, R. Heterologous interaction between β2-microglobulin and Ag-B antigens. Transplantation 30: 233–235, 1980

    Google Scholar 

  • Mauxin, F. and Kress, M. Nucleotide sequence of rat β2-microglobulin cDNA. Nucleic Acids Res 15: 7638, 1987

    Google Scholar 

  • Nakamuro, K., Tanigaki, N., and Pressman, D. Multiple common properties of human β2-microglobulin and the common portion fragment derived from HL-A antigen molecules. Proc Natl Acad Sci USA 70: 2863–2865, 1973

    Google Scholar 

  • Olmo, E., Capriglione, T., and Odierna, G. Genome size in vertebrates: Trends and constraints. Comp Biochem Physiol 92B: 447–453, 1989

    Google Scholar 

  • Ono, H., Klein, D., Vincek, V., Figueroa, F., O'hUigin, C., Tichy, H., and Klein, J. Major histocompatibility complex class II genes of zebrafish. Proc Natl Acad Sci USA 89: 11886–11890, 1992

    Google Scholar 

  • Parnes, J. R. and Seidman, J. G. Structure of wild-type and mutant mouse β2-microglobulin genes. Cell 29: 661–669, 1982

    Google Scholar 

  • Peterson, P. A., Cunningham, B. A., Berggård, I., and Edelman, G. M. β2-microglobulin — a free immunoglobulin domain. Proc Natl Acad Sci USA 69: 1697–1701, 1972

    Google Scholar 

  • Ploegh, H. L., Cannon, L. E., and Strominger, J. L. Cell-free translation of the mRNAs for the heavy and light chains of HLA-A and HLA-B antigens. Proc Natl Acad Sci USA 76: 2273–2277, 1979

    Google Scholar 

  • Ploegh, H. L., Orr, H. T., and Strominger, J. L. Major histocompatibility antigens: the human HLA-A,-B,-C and murine (H-2K, H-2D) class I molecules. Cell 24: 287–299, 1981

    Google Scholar 

  • Rein, R. S., Seemann, G. H. A., Neefjes, J. J., Hochstenbach, F. M. H., Stam, N. J., and Ploegh, H. L. Association with β2-microglobulin controls the expression of transfected human class I genes. J Immunol 138: 1178–1183, 1987

    Google Scholar 

  • Saitou, N. and Nei, M. The neighbor-joining method. A new method for reconstructing phylogenetic trees. Mol Biol Evol 4: 406–425, 1987

    CAS  PubMed  Google Scholar 

  • Sanger, F., Nicklen, S., and Coulson, A. R. DNA sequencing with chain-terminating inhibitors. Proc Natl Acad Sci USA 74: 5463–5467, 1977

    CAS  PubMed  Google Scholar 

  • Saper, M. A., Bjorkman, P. J., and Wiley, D. C. Refined structure of the human histocompatibility antigen HLA-A2 at 2.6 Å resolution. J Mol Biol 219: 277–319, 1991

    CAS  PubMed  Google Scholar 

  • Shalev, A., Greenberg, A. H., Lögdberg, L., and Björck, L. β2-microglobulin-like molecules in low vertebrates and invertebrates. J Immunol 127: 1186–1191, 1981

    Google Scholar 

  • Shalev, A., Pla, M., Ginsburger-Vogel, T., Echalier, G., Lögdberg, E. L., Björck, L., Colombani, J., and Segal, S. Evidence for β2-microglobulin-like and H-2-like antigenic determinants in Drosophila. J Immunol 130: 297–302, 1983

    Google Scholar 

  • Simister, N. E. and Mostov, K. E. An Fc receptor structurally related to MHC class I antigens. Nature 337: 184–187, 1989

    Google Scholar 

  • Smith, R. F. and Smith, T. F. Automatic genration of primary sequence patterns from sets of related protein sequences. Proc Natl Acad Sci USA 87: 118–122, 1990

    Google Scholar 

  • Smithies, O. and Poulik, M. D. Dog homologue of human β2-microglobulin. Proc Natl Acad Sci USA 69: 2914–2917, 1972

    Google Scholar 

  • Tysoe-Calnon, V. A., Grundy, J. E., and Perkins, S. J. Molecular comparisons of the β2-microglobulin-binding site in class I major histocompatibility complex α-chains and proteins of related sequences. Biochem J 277: 359–369, 1991

    Google Scholar 

  • Vincek, V., Sertic, J., Zaleska-Rutczynska, Z., Figueroa, F., and Klein, J. Characterization of 27 new H-2 strains using DNA markers. Immunogenetics 31: 45–51, 1990

    Google Scholar 

  • Warr, G. W., Rosenshein, I. L., and DeLuca, D. β2-microglobulin-like polypeptide of the goldfish, Carassius auratus. Mol Immunol 21: 105–198, 1984

    Google Scholar 

  • Welinder, K. G., Jespersen, H. M., Walther-Rasmussen, J., and Skjødt, K. Amino acid sequences and structure of chicken and turkey beta2-microglobulin. Mol Immunol 28: 177–182, 1991

    Google Scholar 

  • Williams, A. F. and Barclay, A. N. The immunoglobulin superfamily domains for cell surface recognition. Annu Rev Immunol 6: 381–405, 1988

    Google Scholar 

  • Wolfe, P. B. and Cebra, J. J. The primary structure of guinea pig β2-microglobulin. Mol Immunol 17: 1493–1505, 1980

    Google Scholar 

  • Yokoyama, K. and Nathenson, S. G. Intramolecular organization of class I H-2 MHC antigens: Localization of the alloantigenic determinants and the β2m binding site to different regions of the H-2Kb glycoprotein. J Immunol 130: 1419–1425, 1983

    Google Scholar 

  • Zhu, Z., Vincek, V., Figueroa, F., Schönbach, C., and Klein, J. Mhc-DRB genes of the pigtail macaque (Macaca nemestrina): Implications for the evolution of human DRB genes. Mol Biol Evol 8: 563–578, 1991

    Google Scholar 

Download references

Author information

Authors and Affiliations

Authors

Additional information

The nucleotide sequence data reported in this paper have been submitted to the GenBank nucleotide sequence database and have been assigned the accession numbers L05383 (B2M) and L05384 (B2RG).

Correspondence to: J. Klein.

Rights and permissions

Reprints and permissions

About this article

Cite this article

Ono, H., Figueroa, F., O'hUigin, C. et al. Cloning of the β2-microglobulin gene in the zebrafish. Immunogenetics 38, 1–10 (1993). https://doi.org/10.1007/BF00216384

Download citation

  • Received:

  • Revised:

  • Issue Date:

  • DOI: https://doi.org/10.1007/BF00216384

Keywords

Navigation