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. 1991 May 1;173(5):1065–1072. doi: 10.1084/jem.173.5.1065

Deletions of immunoglobulin C kappa region characterized by the circular excision products in mouse splenocytes

PMCID: PMC2118853  PMID: 1902500

Abstract

We have identified circular DNAs containing the kappa light chain constant region (C kappa), as well as the excision products of V kappa- J kappa and V lambda-J lambda joining in adult mouse splenocytes. Analysis of C kappa-positive circular DNA clones revealed two recombination sites (intron recombining sequence [IRS]1 and -2) within the germline J kappa-C kappa intron region and the recombining sequence (RS) located downstream of the C kappa exon. While RS contains a conserved heptamer and nonamer separated by a 23-bp spacer on the 5' side, IRS1 sequence is an isolated heptamer without an obvious nonamer, and IRS2 contains a variant heptamer, CACAAAA. Since IRS1 and IRS2 recombined with both RS (23-bp spacer signal) and V kappa (12-bp spacer signal) with significant frequency, intron recombination sites seem to have dual recombination signals. These findings provide direct evidence that C kappa deletion preceding lambda gene rearrangement can occur by looping out and excision. Increased accessibility of inefficient recombinational loci within the intron may enable recombinase to accept wide signal sequence variation.

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Selected References

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  1. Bernard O., Hozumi N., Tonegawa S. Sequences of mouse immunoglobulin light chain genes before and after somatic changes. Cell. 1978 Dec;15(4):1133–1144. doi: 10.1016/0092-8674(78)90041-7. [DOI] [PubMed] [Google Scholar]
  2. Boyd R. T., Goldrick M. M., Gottlieb P. D. Structural differences in a single gene encoding the V kappa Ser group of light chains explain the existence of two mouse light-chain genetic markers. Proc Natl Acad Sci U S A. 1986 Dec;83(23):9134–9138. doi: 10.1073/pnas.83.23.9134. [DOI] [PMC free article] [PubMed] [Google Scholar]
  3. Campbell M. J., Carroll W., Kon S., Thielemans K., Rothbard J. B., Levy S., Levy R. Idiotype vaccination against murine B cell lymphoma. Humoral and cellular responses elicited by tumor-derived immunoglobulin M and its molecular subunits. J Immunol. 1987 Oct 15;139(8):2825–2833. [PubMed] [Google Scholar]
  4. D'Hoostelaere L. A., Huppi K., Mock B., Mallett C., Potter M. The Ig kappa L chain allelic groups among the Ig kappa haplotypes and Ig kappa crossover populations suggest a gene order. J Immunol. 1988 Jul 15;141(2):652–661. [PubMed] [Google Scholar]
  5. Durdik J., Moore M. W., Selsing E. Novel kappa light-chain gene rearrangements in mouse lambda light chain-producing B lymphocytes. Nature. 1984 Feb 23;307(5953):749–752. doi: 10.1038/307749a0. [DOI] [PubMed] [Google Scholar]
  6. Early P., Huang H., Davis M., Calame K., Hood L. An immunoglobulin heavy chain variable region gene is generated from three segments of DNA: VH, D and JH. Cell. 1980 Apr;19(4):981–992. doi: 10.1016/0092-8674(80)90089-6. [DOI] [PubMed] [Google Scholar]
  7. Feddersen R. M., Martin D. J., Van Ness B. G. Novel recombinations of the IG kappa-locus that result in allelic exclusion. J Immunol. 1990 Jul 15;145(2):745–750. [PubMed] [Google Scholar]
  8. Fujimoto S., Yamagishi H. Isolation of an excision product of T-cell receptor alpha-chain gene rearrangements. Nature. 1987 May 21;327(6119):242–243. doi: 10.1038/327242a0. [DOI] [PubMed] [Google Scholar]
  9. Harada K., Yamagishi H. Lack of feedback inhibition of V kappa gene rearrangement by productively rearranged alleles. J Exp Med. 1991 Feb 1;173(2):409–415. doi: 10.1084/jem.173.2.409. [DOI] [PMC free article] [PubMed] [Google Scholar]
  10. Heinrich G., Traunecker A., Tonegawa S. Somatic mutation creates diversity in the major group of mouse immunoglobulin kappa light chains. J Exp Med. 1984 Feb 1;159(2):417–435. doi: 10.1084/jem.159.2.417. [DOI] [PMC free article] [PubMed] [Google Scholar]
  11. Heller M., Owens J. D., Mushinski J. F., Rudikoff S. Amino acids at the site of V kappa-J kappa recombination not encoded by germline sequences. J Exp Med. 1987 Sep 1;166(3):637–646. doi: 10.1084/jem.166.3.637. [DOI] [PMC free article] [PubMed] [Google Scholar]
  12. Hesse J. E., Lieber M. R., Mizuuchi K., Gellert M. V(D)J recombination: a functional definition of the joining signals. Genes Dev. 1989 Jul;3(7):1053–1061. doi: 10.1101/gad.3.7.1053. [DOI] [PubMed] [Google Scholar]
  13. Hirama T., Takeshita S., Yoshida Y., Yamagishi H. Structure of extrachromosomal circular DNAs generated by immunoglobulin light chain gene rearrangements. Immunol Lett. 1991 Jan;27(1):19–23. doi: 10.1016/0165-2478(91)90238-6. [DOI] [PubMed] [Google Scholar]
  14. Höchtl J., Müller C. R., Zachau H. G. Recombined flanks of the variable and joining segments of immunoglobulin genes. Proc Natl Acad Sci U S A. 1982 Mar;79(5):1383–1387. doi: 10.1073/pnas.79.5.1383. [DOI] [PMC free article] [PubMed] [Google Scholar]
  15. Iwasato T., Shimizu A., Honjo T., Yamagishi H. Circular DNA is excised by immunoglobulin class switch recombination. Cell. 1990 Jul 13;62(1):143–149. doi: 10.1016/0092-8674(90)90248-d. [DOI] [PubMed] [Google Scholar]
  16. Joho R., Gershenfeld H., Weissman I. L. Evolution of a multigene family of V kappa germ line genes. EMBO J. 1984 Jan;3(1):185–191. doi: 10.1002/j.1460-2075.1984.tb01782.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  17. Kaushik A., Schulze D. H., Bona C., Kelsoe G. Murine V kappa gene expression does not follow the VH paradigm. J Exp Med. 1989 May 1;169(5):1859–1864. doi: 10.1084/jem.169.5.1859. [DOI] [PMC free article] [PubMed] [Google Scholar]
  18. Kawakami T., Takahashi N., Honjo T. Complete nucleotide sequence of mouse immunoglobulin mu gene and comparison with other immunoglobulin heavy chain genes. Nucleic Acids Res. 1980 Sep 11;8(17):3933–3945. doi: 10.1093/nar/8.17.3933. [DOI] [PMC free article] [PubMed] [Google Scholar]
  19. Kelley D. E., Wiedemann L. M., Pittet A. C., Strauss S., Nelson K. J., Davis J., Van Ness B., Perry R. P. Nonproductive kappa immunoglobulin genes: recombinational abnormalities and other lesions affecting transcription, RNA processing, turnover, and translation. Mol Cell Biol. 1985 Jul;5(7):1660–1675. doi: 10.1128/mcb.5.7.1660. [DOI] [PMC free article] [PubMed] [Google Scholar]
  20. Kleinfield R., Hardy R. R., Tarlinton D., Dangl J., Herzenberg L. A., Weigert M. Recombination between an expressed immunoglobulin heavy-chain gene and a germline variable gene segment in a Ly 1+ B-cell lymphoma. 1986 Aug 28-Sep 3Nature. 322(6082):843–846. doi: 10.1038/322843a0. [DOI] [PubMed] [Google Scholar]
  21. Kobayashi M., Koike K. Construction and analysis of recombinant lambda phages containing mitochondrial DNA fragments. Gene. 1979 Jun;6(2):123–136. doi: 10.1016/0378-1119(79)90067-2. [DOI] [PubMed] [Google Scholar]
  22. Kurosawa Y., Tonegawa S. Organization, structure, and assembly of immunoglobulin heavy chain diversity DNA segments. J Exp Med. 1982 Jan 1;155(1):201–218. doi: 10.1084/jem.155.1.201. [DOI] [PMC free article] [PubMed] [Google Scholar]
  23. Lawler A. M., Kearney J. F., Kuehl M., Gearhart P. J. Early rearrangements of genes encoding murine immunoglobulin kappa chains, unlike genes encoding heavy chains, use variable gene segments dispersed throughout the locus. Proc Natl Acad Sci U S A. 1989 Sep;86(17):6744–6747. doi: 10.1073/pnas.86.17.6744. [DOI] [PMC free article] [PubMed] [Google Scholar]
  24. Leahy D. J., Rule G. S., Whittaker M. M., McConnell H. M. Sequences of 12 monoclonal anti-dinitrophenyl spin-label antibodies for NMR studies. Proc Natl Acad Sci U S A. 1988 Jun;85(11):3661–3665. doi: 10.1073/pnas.85.11.3661. [DOI] [PMC free article] [PubMed] [Google Scholar]
  25. Lewis S. M., Hesse J. E., Mizuuchi K., Gellert M. Novel strand exchanges in V(D)J recombination. Cell. 1988 Dec 23;55(6):1099–1107. doi: 10.1016/0092-8674(88)90254-1. [DOI] [PubMed] [Google Scholar]
  26. Matsuda T., Kabat E. A. Variable region cDNA sequences and antigen binding specificity of mouse monoclonal antibodies to isomaltosyl oligosaccharides coupled to proteins. T-dependent analogues of alpha(1----6)dextran. J Immunol. 1989 Feb 1;142(3):863–870. [PubMed] [Google Scholar]
  27. Matsuoka M., Yoshida K., Maeda T., Usuda S., Sakano H. Switch circular DNA formed in cytokine-treated mouse splenocytes: evidence for intramolecular DNA deletion in immunoglobulin class switching. Cell. 1990 Jul 13;62(1):135–142. doi: 10.1016/0092-8674(90)90247-c. [DOI] [PubMed] [Google Scholar]
  28. Meyer K. B., Neuberger M. S. The immunoglobulin kappa locus contains a second, stronger B-cell-specific enhancer which is located downstream of the constant region. EMBO J. 1989 Jul;8(7):1959–1964. doi: 10.1002/j.1460-2075.1989.tb03601.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  29. Moore M. W., Durdik J., Persiani D. M., Selsing E. Deletions of kappa chain constant region genes in mouse lambda chain-producing B cells involve intrachromosomal DNA recombinations similar to V-J joining. Proc Natl Acad Sci U S A. 1985 Sep;82(18):6211–6215. doi: 10.1073/pnas.82.18.6211. [DOI] [PMC free article] [PubMed] [Google Scholar]
  30. Ng K. H., Lavigueur A., Ricard L., Boivrette M., Maclean S., Cloutier D., Gibson D. M. Characterization of allelic V kappa-1 region genes in inbred strains of mice. J Immunol. 1989 Jul 15;143(2):638–648. [PubMed] [Google Scholar]
  31. Nishioka Y., Leder P. Organization and complete sequence of identical embryonic and plasmacytoma kappa V-region genes. J Biol Chem. 1980 Apr 25;255(8):3691–3694. [PubMed] [Google Scholar]
  32. Okazaki K., Davis D. D., Sakano H. T cell receptor beta gene sequences in the circular DNA of thymocyte nuclei: direct evidence for intramolecular DNA deletion in V-D-J joining. Cell. 1987 May 22;49(4):477–485. doi: 10.1016/0092-8674(87)90450-8. [DOI] [PubMed] [Google Scholar]
  33. Okazaki K., Sakano H. Thymocyte circular DNA excised from T cell receptor alpha-delta gene complex. EMBO J. 1988 Jun;7(6):1669–1674. doi: 10.1002/j.1460-2075.1988.tb02994.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  34. Pech M., Höchtl J., Schnell H., Zachau H. G. Differences between germ-line and rearranged immunoglobulin V kappa coding sequences suggest a localized mutation mechanism. Nature. 1981 Jun 25;291(5817):668–670. doi: 10.1038/291668a0. [DOI] [PubMed] [Google Scholar]
  35. Persiani D. M., Durdik J., Selsing E. Active lambda and kappa antibody gene rearrangement in Abelson murine leukemia virus-transformed pre-B cell lines. J Exp Med. 1987 Jun 1;165(6):1655–1674. doi: 10.1084/jem.165.6.1655. [DOI] [PMC free article] [PubMed] [Google Scholar]
  36. Reth M., Gehrmann P., Petrac E., Wiese P. A novel VH to VHDJH joining mechanism in heavy-chain-negative (null) pre-B cells results in heavy-chain production. 1986 Aug 28-Sep 3Nature. 322(6082):840–842. doi: 10.1038/322840a0. [DOI] [PubMed] [Google Scholar]
  37. Riley S. C., Connors S. J., Klinman N. R., Ogata R. T. Preferential expression of variable region heavy chain gene segments by predominant 2,4-dinitrophenyl-specific BALB/c neonatal antibody clonotypes. Proc Natl Acad Sci U S A. 1986 Apr;83(8):2589–2593. doi: 10.1073/pnas.83.8.2589. [DOI] [PMC free article] [PubMed] [Google Scholar]
  38. Sakano H., Hüppi K., Heinrich G., Tonegawa S. Sequences at the somatic recombination sites of immunoglobulin light-chain genes. Nature. 1979 Jul 26;280(5720):288–294. doi: 10.1038/280288a0. [DOI] [PubMed] [Google Scholar]
  39. Sakano H., Maki R., Kurosawa Y., Roeder W., Tonegawa S. Two types of somatic recombination are necessary for the generation of complete immunoglobulin heavy-chain genes. Nature. 1980 Aug 14;286(5774):676–683. doi: 10.1038/286676a0. [DOI] [PubMed] [Google Scholar]
  40. Sanger F. Determination of nucleotide sequences in DNA. Science. 1981 Dec 11;214(4526):1205–1210. doi: 10.1126/science.7302589. [DOI] [PubMed] [Google Scholar]
  41. Schnell H., Steinmetz M., Zachau H. G., Schechter I. An unusual translocation of immunoglobulin gene segments in variants of the mouse myeloma MPC11. Nature. 1980 Jul 10;286(5769):170–173. doi: 10.1038/286170a0. [DOI] [PubMed] [Google Scholar]
  42. Seidman J. G., Leder P. A mutant immunoglobulin light chain is formed by aberrant DNA- and RNA-splicing events. Nature. 1980 Aug 21;286(5775):779–783. doi: 10.1038/286779a0. [DOI] [PubMed] [Google Scholar]
  43. Shapiro M. A., Weigert M. How immunoglobulin V kappa genes rearrange. J Immunol. 1987 Dec 1;139(11):3834–3839. [PubMed] [Google Scholar]
  44. Sun L. K., Curtis P., Rakowicz-Szulczynska E., Ghrayeb J., Chang N., Morrison S. L., Koprowski H. Chimeric antibody with human constant regions and mouse variable regions directed against carcinoma-associated antigen 17-1A. Proc Natl Acad Sci U S A. 1987 Jan;84(1):214–218. doi: 10.1073/pnas.84.1.214. [DOI] [PMC free article] [PubMed] [Google Scholar]
  45. Takeshita S., Toda M., Yamagishi H. Excision products of the T cell receptor gene support a progressive rearrangement model of the alpha/delta locus. EMBO J. 1989 Nov;8(11):3261–3270. doi: 10.1002/j.1460-2075.1989.tb08486.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  46. Toda M., Fujimoto S., Iwasato T., Takeshita S., Tezuka K., Ohbayashi T., Yamagishi H. Structure of extrachromosomal circular DNAs excised from T-cell antigen receptor alpha and delta-chain loci. J Mol Biol. 1988 Jul 20;202(2):219–231. doi: 10.1016/0022-2836(88)90453-6. [DOI] [PubMed] [Google Scholar]
  47. Toda M., Hirama T., Takeshita S., Yamagishi H. Excision products of immunoglobulin gene rearrangements. Immunol Lett. 1989 Jun 15;21(4):311–316. doi: 10.1016/0165-2478(89)90025-4. [DOI] [PubMed] [Google Scholar]
  48. Van Cleave V. H., Naeve C. W., Metzger D. W. Do antibodies recognize amino acid side chains of protein antigens independently of the carbon backbone? J Exp Med. 1988 Jun 1;167(6):1841–1848. doi: 10.1084/jem.167.6.1841. [DOI] [PMC free article] [PubMed] [Google Scholar]
  49. Yancopoulos G. D., Alt F. W. Regulation of the assembly and expression of variable-region genes. Annu Rev Immunol. 1986;4:339–368. doi: 10.1146/annurev.iy.04.040186.002011. [DOI] [PubMed] [Google Scholar]
  50. de Villartay J. P., Hockett R. D., Coran D., Korsmeyer S. J., Cohen D. I. Deletion of the human T-cell receptor delta-gene by a site-specific recombination. Nature. 1988 Sep 8;335(6186):170–174. doi: 10.1038/335170a0. [DOI] [PubMed] [Google Scholar]
  51. von Schwedler U., Jäck H. M., Wabl M. Circular DNA is a product of the immunoglobulin class switch rearrangement. Nature. 1990 May 31;345(6274):452–456. doi: 10.1038/345452a0. [DOI] [PubMed] [Google Scholar]

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