Cloning and Characterization of a Protein Binding to the Jκ Recombination Signal Sequence of Immunoglobulin Genes

  • Yasushi Hamaguchi
  • Norisada Mastunami
  • Yoshiki Yamamoto
  • Kogo Kuze
  • Kenji Kangawa
  • Hisayuki Matsuo
  • Masashi Kawaichi
  • Tasuku Honjo
Part of the Advances in Experimental Medicine and Biology book series (AEMB, volume 292)


A protein with molecular weight of 60,000 that binds to the recombination signal sequence (RS) of the immunoglobulin Jκ segment was purified from the nuclear extract of a murine pre B cell line 38B9. This binding protein was found in lymphoid cell lines but not in non-lymphoid cell lines. The Kd value of the Jκ RS binding protein to the Jκ RS was 1 nM. The cDNA clone (RBP-2) was isolated based on partial amino-acid sequence of this protein. This cDNA encodes 526 amino-acid residues, and its sequence does not show extensive overall homology with any known proteins, but displays an interesting homology to a 40-residue region that is conserved among a subset of site specific recombinase (integrase family).


Immunoglobulin Gene Partial Amino Acid Sequence Lymphoid Cell Line Variable Region Gene Cell Receptor Gene 
These keywords were added by machine and not by the authors. This process is experimental and the keywords may be updated as the learning algorithm improves.


  1. 1.
    P. Early, H. Huang, M. Davis, K. Calame, and L. Hood, An immunoglobulin heavy chain variable region gene is generated from three segments of DNA: VH, D and JH, Cell 19:981 (1980).PubMedCrossRefGoogle Scholar
  2. 2.
    H. Sakano, R. Maki, Y. Kurosawa, W. Roeder, M. Weigert, and S. Tonegawa, Two types of somatic recombinations are necessary for the generation of complete immunoglobulin heavy chain genes, Nature 286:676 (1980).PubMedCrossRefGoogle Scholar
  3. 3.
    S. Lewis, A. Gifford, and D. Baltimore, Joining of Va. to Ja. gene segments in a retroviral vector introduced into lymphoid cells, Nature 286:676 (1980).CrossRefGoogle Scholar
  4. 4.
    T. K. Blackwell and F. W. Alt, Site-specific recombination between immunoglobulin D and JR segments that were introduced into the genome of a murine pre-B cell line, Cell 37:105 (1984).PubMedCrossRefGoogle Scholar
  5. 5.
    S. Akira, K. Okazaki, and H. Sakano, Two pairs of recombination signals are sufficient to cause immunoglobulin V-(D)-J joining, Science 238:1134 (1987).PubMedCrossRefGoogle Scholar
  6. 6.
    F. Alt, N. Rosenberg, S. Lewis, E. Thomas, and D. Baltimore, Organization and reorganization of immunoglobulin genes in A-MuLV-transformed cells: rearrangement of heavy but not light chain genes, Cell 27:381 (1981).PubMedCrossRefGoogle Scholar
  7. 7.
    Y. Hamaguchi, N. Matsunami, Y. Yamamoto, and T. Honjo, Purification and characterization of a protein that binds to the recombination signal sequence of the immunoglobulin Jk segment, Nucleic Acids Res. 17:9015 (1989).PubMedCrossRefGoogle Scholar
  8. 8.
    R. Palacios and M. Steinmetz, IL3-dependent mouse clones that express B-220 surface antigen, contain Ig genes in germ-line configuration, and generate B lymphocytes in vivo, Cell 41:727 (1985).PubMedCrossRefGoogle Scholar
  9. 9.
    J. Pelkonen, P. Sideras, H.-G. Rammensee, K. Karjalainen, and R. Palacios, Thymocyte clones from 14-day mouse embryos. I. State of T cell receptor genes, surface markers, and growth requirements, J. Exp. Med. 166:1245 (1987).PubMedCrossRefGoogle Scholar
  10. 10.
    N. Matsunami, Y. Hamaguchi, Y. Yamanoto, K. Kuze, K. Kangawa, H. Matsuo, M. Kawaichi, and T. Honjo, A protein binding to the Ja. recombination sequence of immunoglobulin genes contains a sequence rerated to the integrase motif, Nature 342:934 (1989).PubMedCrossRefGoogle Scholar
  11. 11.
    P. Argos, A. Landy, K. Abremski, J. B. Egan, E. Haggard-Ljungquist, R. H. Hoess, M. L. Kahn, B. Kalionis, S. V. L. Narayana, L. S. Pierson, N. Sternberg, and J. M. Leong, The integrase family of site-specific recombinases: regional similarities and global diversity, EMBO J. 5:433 (1986).PubMedGoogle Scholar
  12. 12.
    G. Yancopoulos, T. K. Blackwell, H. Suh, L. Hood, and F. W. Alt, Introduced T cell receptor variable region gene segments recombine in pre-B cells: evidence that B and T cells use a common recombinase, Cell 44:251 (1986).PubMedCrossRefGoogle Scholar
  13. 13.
    J. Kavaler, M. M. Davis, and Y. Chien, Localization of a T-cell receptor diversity-region element, Nature 310:421 (1984).PubMedCrossRefGoogle Scholar
  14. 14.
    E. A. Kabat, T. T. Wu, M. Reid-Miller, H. M. Perry, and K. S. Gottesman, “Sequences of Proteins of Immunological Interest, Ed. 4.” NIH, Bethesda (1987).Google Scholar
  15. 15.
    N. L. Craig, The mechanism of conservative site-specific recombination, Annu. Rev. Genet. 22:77 (1988).PubMedCrossRefGoogle Scholar
  16. 16.
    R. L. Parsons, P. V. Prasad, R. M. Harshey, and M. Jayaram, Step-arrest mutants of FLP recombinase: implications for the catalytic mechanism of DNA recombination. Mol. Cell. Biol. 8:3303 (1988).PubMedGoogle Scholar
  17. 17.
    P. V. Prasad, L.-J. Young, and M. Jayaram, Mutations in the 2-m circle site-specific recombinase that abolish recombination without affecting substrate recognition, Proc. Natl. Acad. Sci. USA 84:2189 (1987).PubMedCrossRefGoogle Scholar
  18. 18.
    C. A. Pargellis, S. E. Nunes-Düby, L. Moitoso de Vargas, and A. Landy, Suicide recombination substrates yield covalent integrase-DNA complexes and lead to identification of the active site tyrosine, J. Biol. Chem. 263:7678 (1988).PubMedGoogle Scholar
  19. 19.
    L. S. Pierson, and M. L. Kahn, Integration of satellite bacteriophage P4 in Escherichia colt DNA sequences of the phage and host regions involved in site-specific recombination. J. Mol. Biol. 196:487 (1987).PubMedCrossRefGoogle Scholar
  20. 20.
    R. J. Aguilera, S. Akira, K. Okazaki, and H. Sakano, A pre-B cell nuclear protein which specifically interacts with the immunoglobulin V-J recombination sequences. Cell 51:909 (1987).PubMedCrossRefGoogle Scholar
  21. 21.
    B. D. Halligan and S. V. Desiderio, Identification of a DNA binding protein that recognizes the nonamer recombinational signal sequence of immunoglobulin genes, Proc. Natl. Acad. Sci. USA 84:7019 (1987).PubMedCrossRefGoogle Scholar

Copyright information

© Plenum Press, New York 1991

Authors and Affiliations

  • Yasushi Hamaguchi
    • 1
  • Norisada Mastunami
    • 1
  • Yoshiki Yamamoto
    • 1
  • Kogo Kuze
    • 1
  • Kenji Kangawa
    • 2
  • Hisayuki Matsuo
    • 2
  • Masashi Kawaichi
    • 1
  • Tasuku Honjo
    • 1
  1. 1.Department of Medical Chemistry, Faculty of MedicineKyoto UniversityKyotoJapan
  2. 2.Department of BiochemistryMiyazaki Medical CollegeMiyazakiJapan

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