Skip to main content
The EMBO Journal logoLink to The EMBO Journal
. 1990 Aug;9(8):2501–2506. doi: 10.1002/j.1460-2075.1990.tb07429.x

Organization of variable region segments of the human immunoglobulin heavy chain: duplication of the D5 cluster within the locus and interchromosomal translocation of variable region segments.

F Matsuda 1, E K Shin 1, Y Hirabayashi 1, H Nagaoka 1, M C Yoshida 1, S Q Zong 1, T Honjo 1
PMCID: PMC552279  PMID: 2114977

Abstract

We have studied the organization of variable region (V) genes of the human immunoglobulin heavy chain (H) by cosmid cloning. We isolated two independent immunoglobulin D5 clusters (D5-a and D5-b) from cosmid libraries of the human genome. Restriction maps of these two regions showed that downstream 15 kb portions of the 55 kb overlap were different although upstream 40 kb portions were almost identical. Four more D segments, (DM, DXP, DA and DK) were found around the D5 segment in the conserved region of each cluster. Nucleotide sequences of the corresponding D segments from each cluster were almost identical and they encoded potentially functional D regions. Analysis using human-rodent somatic cell hybrids demonstrated that both clusters were located in the immunoglobulin heavy chain (H) locus on chromosome 14, suggesting that the D5-a and D5-b regions evolved by internal duplication within this locus. We also isolated a 60 kb DNA region carrying four VH segments, designated as VH-F region, which was located on chromosome 16. Nucleotide sequences of the four VH segments were determined. Two of them encoded potentially functional VH segments, and the other two were pseudogenes. Some more VH segments were found to be located outside chromosome 14, by Southern blot hybridization of human-rodent hybrid cell DNAs. These results provide further evidence that the human VH locus has undergone recent reorganization.

Full text

PDF
2501

Images in this article

Selected References

These references are in PubMed. This may not be the complete list of references from this article.

  1. Akira S., Okazaki K., Sakano H. Two pairs of recombination signals are sufficient to cause immunoglobulin V-(D)-J joining. Science. 1987 Nov 20;238(4830):1134–1138. doi: 10.1126/science.3120312. [DOI] [PubMed] [Google Scholar]
  2. Berman J. E., Mellis S. J., Pollock R., Smith C. L., Suh H., Heinke B., Kowal C., Surti U., Chess L., Cantor C. R. Content and organization of the human Ig VH locus: definition of three new VH families and linkage to the Ig CH locus. EMBO J. 1988 Mar;7(3):727–738. doi: 10.1002/j.1460-2075.1988.tb02869.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  3. Brodeur P. H., Osman G. E., Mackle J. J., Lalor T. M. The organization of the mouse Igh-V locus. Dispersion, interspersion, and the evolution of VH gene family clusters. J Exp Med. 1988 Dec 1;168(6):2261–2278. doi: 10.1084/jem.168.6.2261. [DOI] [PMC free article] [PubMed] [Google Scholar]
  4. Croce C. M., Shander M., Martinis J., Cicurel L., D'Ancona G. G., Dolby T. W., Koprowski H. Chromosomal location of the genes for human immunoglobulin heavy chains. Proc Natl Acad Sci U S A. 1979 Jul;76(7):3416–3419. doi: 10.1073/pnas.76.7.3416. [DOI] [PMC free article] [PubMed] [Google Scholar]
  5. Gunning P., Ponte P., Kedes L., Hickey R. J., Skoultchi A. I. Expression of human cardiac actin in mouse L cells: a sarcomeric actin associates with a nonmuscle cytoskeleton. Cell. 1984 Mar;36(3):709–715. doi: 10.1016/0092-8674(84)90351-9. [DOI] [PubMed] [Google Scholar]
  6. Hattori M., Sakaki Y. Dideoxy sequencing method using denatured plasmid templates. Anal Biochem. 1986 Feb 1;152(2):232–238. doi: 10.1016/0003-2697(86)90403-3. [DOI] [PubMed] [Google Scholar]
  7. Honjo T., Habu S. Origin of immune diversity: genetic variation and selection. Annu Rev Biochem. 1985;54:803–830. doi: 10.1146/annurev.bi.54.070185.004103. [DOI] [PubMed] [Google Scholar]
  8. Humphries C. G., Shen A., Kuziel W. A., Capra J. D., Blattner F. R., Tucker P. W. A new human immunoglobulin VH family preferentially rearranged in immature B-cell tumours. Nature. 1988 Feb 4;331(6155):446–449. doi: 10.1038/331446a0. [DOI] [PubMed] [Google Scholar]
  9. Ichihara Y., Abe M., Yasui H., Matsuoka H., Kurosawa Y. At least five DH genes of human immunoglobulin heavy chains are encoded in 9-kilobase DNA fragments. Eur J Immunol. 1988 Apr;18(4):649–652. doi: 10.1002/eji.1830180426. [DOI] [PubMed] [Google Scholar]
  10. Ichihara Y., Hayashida H., Miyazawa S., Kurosawa Y. Only DFL16, DSP2, and DQ52 gene families exist in mouse immunoglobulin heavy chain diversity gene loci, of which DFL16 and DSP2 originate from the same primordial DH gene. Eur J Immunol. 1989 Oct;19(10):1849–1854. doi: 10.1002/eji.1830191014. [DOI] [PubMed] [Google Scholar]
  11. Ichihara Y., Matsuoka H., Kurosawa Y. Organization of human immunoglobulin heavy chain diversity gene loci. EMBO J. 1988 Dec 20;7(13):4141–4150. doi: 10.1002/j.1460-2075.1988.tb03309.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  12. Katamine S., Otsu M., Tada K., Tsuchiya S., Sato T., Ishida N., Honjo T., Ono Y. Epstein-Barr virus transforms precursor B cells even before immunoglobulin gene rearrangements. Nature. 1984 May 24;309(5966):369–372. doi: 10.1038/309369a0. [DOI] [PubMed] [Google Scholar]
  13. Kodaira M., Kinashi T., Umemura I., Matsuda F., Noma T., Ono Y., Honjo T. Organization and evolution of variable region genes of the human immunoglobulin heavy chain. J Mol Biol. 1986 Aug 20;190(4):529–541. doi: 10.1016/0022-2836(86)90239-1. [DOI] [PubMed] [Google Scholar]
  14. Lee K. H., Matsuda F., Kinashi T., Kodaira M., Honjo T. A novel family of variable region genes of the human immunoglobulin heavy chain. J Mol Biol. 1987 Jun 20;195(4):761–768. doi: 10.1016/0022-2836(87)90482-7. [DOI] [PubMed] [Google Scholar]
  15. Lorenz W., Straubinger B., Zachau H. G. Physical map of the human immunoglobulin K locus and its implications for the mechanisms of VK-JK rearrangement. Nucleic Acids Res. 1987 Dec 10;15(23):9667–9676. doi: 10.1093/nar/15.23.9667. [DOI] [PMC free article] [PubMed] [Google Scholar]
  16. Lötscher E., Zimmer F. J., Klopstock T., Grzeschik K. H., Jaenichen R., Straubinger B., Zachau H. G. Localization, analysis and evolution of transposed human immunoglobulin V kappa genes. Gene. 1988 Sep 30;69(2):215–223. doi: 10.1016/0378-1119(88)90432-5. [DOI] [PubMed] [Google Scholar]
  17. Maeda T., Sugiyama H., Tani Y., Miyake S., Oka Y., Ogawa H., Komori T., Soma T., Kishimoto S. Start of mu-chain production by the further two-step rearrangements of immunoglobulin heavy chain genes on one chromosome from a DJH/DJH configuration in an Abelson virus-transformed cell line: evidence of secondary DJH complex formation. J Immunol. 1987 Apr 1;138(7):2305–2310. [PubMed] [Google Scholar]
  18. Matsuda F., Lee K. H., Nakai S., Sato T., Kodaira M., Zong S. Q., Ohno H., Fukuhara S., Honjo T. Dispersed localization of D segments in the human immunoglobulin heavy-chain locus. EMBO J. 1988 Apr;7(4):1047–1051. doi: 10.1002/j.1460-2075.1988.tb02912.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  19. Nakamura Y., Martin C., Krapcho K., O'Connell P., Leppert M., Lathrop G. M., Lalouel J. M., White R. Isolation and mapping of a polymorphic DNA sequence (pCMM65) on chromosome 16 [D16S84]. Nucleic Acids Res. 1988 Apr 11;16(7):3122–3122. doi: 10.1093/nar/16.7.3122. [DOI] [PMC free article] [PubMed] [Google Scholar]
  20. Nishida Y., Miki T., Hisajima H., Honjo T. Cloning of human immunoglobulin epsilon chain genes: evidence for multiple C epsilon genes. Proc Natl Acad Sci U S A. 1982 Jun;79(12):3833–3837. doi: 10.1073/pnas.79.12.3833. [DOI] [PMC free article] [PubMed] [Google Scholar]
  21. Otsu M., Katamine S., Uno M., Yamaki M., Ono Y., Klein G., Sasaki M. S., Yaoita Y., Honjo T. Molecular characterization of novel reciprocal translocation t(6;14) in an Epstein-Barr virus-transformed B cell precursor. Mol Cell Biol. 1987 Feb;7(2):708–717. doi: 10.1128/mcb.7.2.708. [DOI] [PMC free article] [PubMed] [Google Scholar]
  22. Purrello M., Alhadeff B., Whittington E., Buckton K. E., Daniel A., Arnaud P., Rocchi M., Archidiacono N., Filippi G., Siniscalco M. Comparison of cytologic and genetic distances between long arm subtelomeric markers of human autosome 14 suggests uneven distribution of crossing-over. Cytogenet Cell Genet. 1987;44(1):32–40. doi: 10.1159/000132337. [DOI] [PubMed] [Google Scholar]
  23. Reth M. G., Jackson S., Alt F. W. VHDJH formation and DJH replacement during pre-B differentiation: non-random usage of gene segments. EMBO J. 1986 Sep;5(9):2131–2138. doi: 10.1002/j.1460-2075.1986.tb04476.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  24. Sanger F., Coulson A. R., Barrell B. G., Smith A. J., Roe B. A. Cloning in single-stranded bacteriophage as an aid to rapid DNA sequencing. J Mol Biol. 1980 Oct 25;143(2):161–178. doi: 10.1016/0022-2836(80)90196-5. [DOI] [PubMed] [Google Scholar]
  25. Schroeder H. W., Jr, Hillson J. L., Perlmutter R. M. Early restriction of the human antibody repertoire. Science. 1987 Nov 6;238(4828):791–793. doi: 10.1126/science.3118465. [DOI] [PubMed] [Google Scholar]
  26. Southern E. M. Detection of specific sequences among DNA fragments separated by gel electrophoresis. J Mol Biol. 1975 Nov 5;98(3):503–517. doi: 10.1016/s0022-2836(75)80083-0. [DOI] [PubMed] [Google Scholar]
  27. Takahashi N., Noma T., Honjo T. Rearranged immunoglobulin heavy chain variable region (VH) pseudogene that deletes the second complementarity-determining region. Proc Natl Acad Sci U S A. 1984 Aug;81(16):5194–5198. doi: 10.1073/pnas.81.16.5194. [DOI] [PMC free article] [PubMed] [Google Scholar]
  28. Tonegawa S. Somatic generation of antibody diversity. Nature. 1983 Apr 14;302(5909):575–581. doi: 10.1038/302575a0. [DOI] [PubMed] [Google Scholar]
  29. Tycko B., Palmer J. D., Sklar J. T cell receptor gene trans-rearrangements: chimeric gamma-delta genes in normal lymphoid tissues. Science. 1989 Sep 15;245(4923):1242–1246. doi: 10.1126/science.2551037. [DOI] [PubMed] [Google Scholar]
  30. Zong S. Q., Nakai S., Matsuda F., Lee K. H., Honjo T. Human immunoglobulin D segments: isolation of a new D segment and polymorphic deletion of the D1 segment. Immunol Lett. 1988 Apr;17(4):329–333. doi: 10.1016/0165-2478(88)90006-5. [DOI] [PubMed] [Google Scholar]

Articles from The EMBO Journal are provided here courtesy of Nature Publishing Group

RESOURCES