Skip to main content
PMC home page
Proceedings of the National Academy of Sciences of the United States of America logoLink to Proceedings of the National Academy of Sciences of the United States of America
. 1984 Aug;81(15):4756–4760. doi: 10.1073/pnas.81.15.4756

Different joining region J elements of the murine kappa immunoglobulin light chain locus are used at markedly different frequencies.

D L Wood, C Coleclough
PMCID: PMC391569  PMID: 6431410

Abstract

In order to assess whether DNA rearrangement occurs with equal frequency at each of the several J (joining region) elements in the mouse kappa light chain locus, we set out to determine the relative frequency of usage of J kappa segments in populations of B lymphocytes unperturbed by antigenic selection or cloning. To obtain such a population, we exposed a suspension of spleen cells to a mixture of mitogens capable of activating most B cells independently of their specificity for antigen. We estimated the relative usage of the J kappa elements in unspliced kappa chain gene transcripts in total and poly(A)-containing nuclear RNA, using an S1 nuclease protection assay, and in mature kappa chain mRNA, using a specifically primed cDNA hybridization assay. Both types of assay reveal a marked difference in the frequency of J kappa elements and indicate that their relative usage is: J1 approximately J2 much greater than J4 approximately J3. Comparison of the 5' flanking regions of the mouse J kappa elements, including the conserved putative recombination target sequences, shows no obvious differences consistent with the variation in recombinational efficiency, so we conclude that, although the consensus heptamer and nonamer signals may be sufficient to identify a recombination site, the probability that that site will be used depends also on other determinants. A review of published data suggests a nonequivalence of usage also among human J kappa elements and between mouse J lambda I and J lambda III loci. Extending the comparison to include these sets of sequences indicates that one of the determinants of frequency of J use may be the proximity of the consensus heptamer to a T-G dinucleotide within the J coding sequence, perhaps revealing an Escherichia coli gyrase-like substrate preference within a recombination enzyme.

Full text

PDF
4756

Images in this article

4757Image
on p.4757
4757Image
on p.4757
4758Image
on p.4758

Selected References

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

  1. Coleclough C. Chance, necessity and antibody gene dynamics. Nature. 1983 May 5;303(5912):23–26. doi: 10.1038/303023a0. [DOI] [PubMed] [Google Scholar]
  2. Coleclough C., Perry R. P., Karjalainen K., Weigert M. Aberrant rearrangements contribute significantly to the allelic exclusion of immunoglobulin gene expression. Nature. 1981 Apr 2;290(5805):372–378. doi: 10.1038/290372a0. [DOI] [PubMed] [Google Scholar]
  3. 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]
  4. Eshhar Z., Blatt C., Bergman Y., Heimovich J. Induction of secretion of IgM from cells of the B cell line 38c-13 by somatic cell hybridization. J Immunol. 1979 Jun;122(6):2430–2434. [PubMed] [Google Scholar]
  5. Hieter P. A., Maizel J. V., Jr, Leder P. Evolution of human immunoglobulin kappa J region genes. J Biol Chem. 1982 Feb 10;257(3):1516–1522. [PubMed] [Google Scholar]
  6. Ikeda H., Aoki K., Naito A. Illegitimate recombination mediated in vitro by DNA gyrase of Escherichia coli: structure of recombinant DNA molecules. Proc Natl Acad Sci U S A. 1982 Jun;79(12):3724–3728. doi: 10.1073/pnas.79.12.3724. [DOI] [PMC free article] [PubMed] [Google Scholar]
  7. Marvo S. L., King S. R., Jaskunas S. R. Role of short regions of homology in intermolecular illegitimate recombination events. Proc Natl Acad Sci U S A. 1983 May;80(9):2452–2456. doi: 10.1073/pnas.80.9.2452. [DOI] [PMC free article] [PubMed] [Google Scholar]
  8. Max E. E., Maizel J. V., Jr, Leder P. The nucleotide sequence of a 5.5-kilobase DNA segment containing the mouse kappa immunoglobulin J and C region genes. J Biol Chem. 1981 May 25;256(10):5116–5120. [PubMed] [Google Scholar]
  9. Max E. E., Seidman J. G., Leder P. Sequences of five potential recombination sites encoded close to an immunoglobulin kappa constant region gene. Proc Natl Acad Sci U S A. 1979 Jul;76(7):3450–3454. doi: 10.1073/pnas.76.7.3450. [DOI] [PMC free article] [PubMed] [Google Scholar]
  10. Miller J., Selsing E., Storb U. Structural alterations in J regions of mouse immunoglobulin lambda genes are associated with differential gene expression. Nature. 1982 Feb 4;295(5848):428–430. doi: 10.1038/295428a0. [DOI] [PubMed] [Google Scholar]
  11. Morrison A., Cozzarelli N. R. Site-specific cleavage of DNA by E. coli DNA gyrase. Cell. 1979 May;17(1):175–184. doi: 10.1016/0092-8674(79)90305-2. [DOI] [PubMed] [Google Scholar]
  12. Perry R. P., Kelley D. E., Coleclough C., Seidman J. G., Leder P., Tonegawa S., Matthyssens G., Weigert M. Transcription of mouse kappa chain genes: implications for allelic exclusion. Proc Natl Acad Sci U S A. 1980 Apr;77(4):1937–1941. doi: 10.1073/pnas.77.4.1937. [DOI] [PMC free article] [PubMed] [Google Scholar]
  13. Reilly E. B., Frackelton A. R., Jr, Eisen H. N. Synthesis of lambda 1, lambda 2, and lambda 3 light chains by mouse spleen B cells. Eur J Immunol. 1982 Jul;12(7):552–557. doi: 10.1002/eji.1830120705. [DOI] [PubMed] [Google Scholar]
  14. Rigby P. W., Dieckmann M., Rhodes C., Berg P. Labeling deoxyribonucleic acid to high specific activity in vitro by nick translation with DNA polymerase I. J Mol Biol. 1977 Jun 15;113(1):237–251. doi: 10.1016/0022-2836(77)90052-3. [DOI] [PubMed] [Google Scholar]
  15. 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]
  16. Sakano H., Kurosawa Y., Weigert M., Tonegawa S. Identification and nucleotide sequence of a diversity DNA segment (D) of immunoglobulin heavy-chain genes. Nature. 1981 Apr 16;290(5807):562–565. doi: 10.1038/290562a0. [DOI] [PubMed] [Google Scholar]
  17. 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]
  18. Schibler U., Marcu K. B., Perry R. P. The synthesis and processing of the messenger RNAs specifying heavy and light chain immunoglobulins in MPC-11 cells. Cell. 1978 Dec;15(4):1495–1509. doi: 10.1016/0092-8674(78)90072-7. [DOI] [PubMed] [Google Scholar]
  19. 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]
  20. Van Ness B. G., Weigert M., Coleclough C., Mather E. L., Kelley D. E., Perry R. P. Transcription of the unrearranged mouse C kappa locus: sequence of the initiation region and comparison of activity with a rearranged V kappa-C kappa gene. Cell. 1981 Dec;27(3 Pt 2):593–602. doi: 10.1016/0092-8674(81)90401-3. [DOI] [PubMed] [Google Scholar]
  21. Wetzel G. D., Kettman J. R. Activation of murine B lymphocytes. III. Stimulation of B lymphocyte clonal growth with lipopolysaccharide and dextran sulfate. J Immunol. 1981 Feb;126(2):723–728. [PubMed] [Google Scholar]
  22. Wood C., Tonegawa S. Diversity and joining segments of mouse immunoglobulin heavy chain genes are closely linked and in the same orientation: implications for the joining mechanism. Proc Natl Acad Sci U S A. 1983 May;80(10):3030–3034. doi: 10.1073/pnas.80.10.3030. [DOI] [PMC free article] [PubMed] [Google Scholar]

Articles from Proceedings of the National Academy of Sciences of the United States of America are provided here courtesy of National Academy of Sciences

RESOURCES