Skip to main content
NCBI 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
. 1987 Feb;84(4):1085–1089. doi: 10.1073/pnas.84.4.1085

V kappa and J kappa gene segments of A/J Ars-A antibodies: somatic recombination generates the essential arginine at the junction of the variable and joining regions.

I Sanz, J D Capra
PMCID: PMC304366  PMID: 3103124

Abstract

The germ-line elements for the variable (V) and joining (J) regions of the kappa chains (V kappa-Ars, J kappa 1) that give rise to the productive allele for the A/J mouse light chains of Ars-A monoclonal antibodies (directed against the hapten p-azophenylarsonate) have been cloned and sequenced as well as a rearranged Ars-A light chain gene. Using the V kappa-Ars gene as a hybridization probe, we provide evidence that the V kappa 10 family is relatively small and that only one member gives rise to the productive allele in all Ars-A antibodies. An unusual feature of Ars-A light chains is that all contain an arginine at position 96, the V-J junctional position. We had shown by chain recombination that arginine-96 was essential for antigen binding. In the present study, an arginine codon was not found either at the 3' end of the V kappa-Ars gene segment or at the 5' end of J kappa 1. However, an arginine codon (CGG) can easily be generated by recombination between these two germ-line elements. Thus, we document that junctional diversity through intracodonic recombination can be crucial to the antibody binding function of the resulting molecule.

Full text

PDF
1085

Images in this article

1088Image
on p.1088
1088Image
on p.1088
1088Image
on p.1088

Selected References

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

  1. Alt F. W., Blackwell T. K., DePinho R. A., Reth M. G., Yancopoulos G. D. Regulation of genome rearrangement events during lymphocyte differentiation. Immunol Rev. 1986 Feb;89:5–30. doi: 10.1111/j.1600-065x.1986.tb01470.x. [DOI] [PubMed] [Google Scholar]
  2. Berek C., Griffiths G. M., Milstein C. Molecular events during maturation of the immune response to oxazolone. Nature. 1985 Aug 1;316(6027):412–418. doi: 10.1038/316412a0. [DOI] [PubMed] [Google Scholar]
  3. Cohn M. Conversations with Niels Kaj Jerne on immune regulation: associative versus network recognition. Cell Immunol. 1981 Jul 1;61(2):425–436. doi: 10.1016/0008-8749(81)90390-7. [DOI] [PubMed] [Google Scholar]
  4. Crews S., Griffin J., Huang H., Calame K., Hood L. A single VH gene segment encodes the immune response to phosphorylcholine: somatic mutation is correlated with the class of the antibody. Cell. 1981 Jul;25(1):59–66. doi: 10.1016/0092-8674(81)90231-2. [DOI] [PubMed] [Google Scholar]
  5. Feinberg A. P., Vogelstein B. A technique for radiolabeling DNA restriction endonuclease fragments to high specific activity. Anal Biochem. 1983 Jul 1;132(1):6–13. doi: 10.1016/0003-2697(83)90418-9. [DOI] [PubMed] [Google Scholar]
  6. Gearhart P. J., Bogenhagen D. F. Clusters of point mutations are found exclusively around rearranged antibody variable genes. Proc Natl Acad Sci U S A. 1983 Jun;80(11):3439–3443. doi: 10.1073/pnas.80.11.3439. [DOI] [PMC free article] [PubMed] [Google Scholar]
  7. Gearhart P. J., Johnson N. D., Douglas R., Hood L. IgG antibodies to phosphorylcholine exhibit more diversity than their IgM counterparts. Nature. 1981 May 7;291(5810):29–34. doi: 10.1038/291029a0. [DOI] [PubMed] [Google Scholar]
  8. Hozumi N., Hawley R. G., Murialdo H. Molecular cloning of an immunoglobulin kappa constant gene from NZB mouse. Gene. 1981 Mar;13(2):163–172. doi: 10.1016/0378-1119(81)90005-6. [DOI] [PubMed] [Google Scholar]
  9. Jerne N. K., Roland J., Cazenave P. A. Recurrent idiotopes and internal images. EMBO J. 1982;1(2):243–247. doi: 10.1002/j.1460-2075.1982.tb01154.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  10. Jeske D. J., Jarvis J., Milstein C., Capra J. D. Junctional diversity is essential to antibody activity. J Immunol. 1984 Sep;133(3):1090–1092. [PubMed] [Google Scholar]
  11. 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]
  12. Kaartinen M., Griffiths G. M., Markham A. F., Milstein C. mRNA sequences define an unusually restricted IgG response to 2-phenyloxazolone and its early diversification. 1983 Jul 28-Aug 3Nature. 304(5924):320–324. doi: 10.1038/304320a0. [DOI] [PubMed] [Google Scholar]
  13. Landolfi N. F., Capra J. D., Tucker P. W. Germ-line sequence of the DH segment employed in Ars-A antibodies: implications for the generation of junctional diversity. J Immunol. 1986 Jul 1;137(1):362–365. [PubMed] [Google Scholar]
  14. Leo O., Urbain J., Moser M., Marvel J., Hiernaux J., Slaoui M. Idiotypic manipulations in the arsonate system. Ann Immunol (Paris) 1984 Jan-Feb;135C(1):45–50. doi: 10.1016/s0769-2625(84)80010-0. [DOI] [PubMed] [Google Scholar]
  15. Milner E. C., Capra J. D. VH families in the antibody response to p-azophenylarsonate: correlation between serology and amino acid sequence. J Immunol. 1982 Jul;129(1):193–199. [PubMed] [Google Scholar]
  16. Perlmutter R. M., Crews S. T., Douglas R., Sorensen G., Johnson N., Nivera N., Gearhart P. J., Hood L. The generation of diversity in phosphorylcholine-binding antibodies. Adv Immunol. 1984;35:1–37. doi: 10.1016/s0065-2776(08)60572-6. [DOI] [PubMed] [Google Scholar]
  17. Rocca-Serra J., Matthes H. W., Kaartinen M., Milstein C., Thèze J., Fougereau M. Analysis of antibody diversity: V-D-J mRNA nucleotide sequence of four anti-GAT monoclonal antibodies. A paucigene system using alternate D-J recombinations to generate functionally similar hypervariable regions. EMBO J. 1983;2(6):867–872. doi: 10.1002/j.1460-2075.1983.tb01515.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  18. 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]
  19. Sanger F., Nicklen S., Coulson A. R. DNA sequencing with chain-terminating inhibitors. Proc Natl Acad Sci U S A. 1977 Dec;74(12):5463–5467. doi: 10.1073/pnas.74.12.5463. [DOI] [PMC free article] [PubMed] [Google Scholar]
  20. Sharon J., Gefter M. L., Manser T., Ptashne M. Site-directed mutagenesis of an invariant amino acid residue at the variable-diversity segments junction of an antibody. Proc Natl Acad Sci U S A. 1986 Apr;83(8):2628–2631. doi: 10.1073/pnas.83.8.2628. [DOI] [PMC free article] [PubMed] [Google Scholar]
  21. Siegelman M., Capra J. D. Complete amino acid sequence of light chain variable regions derived from five monoclonal anti-p-azophenylarsonate antibodies differing with respect to a crossreactive idiotype. Proc Natl Acad Sci U S A. 1981 Dec;78(12):7679–7683. doi: 10.1073/pnas.78.12.7679. [DOI] [PMC free article] [PubMed] [Google Scholar]
  22. Siekevitz M., Huang S. Y., Gefter M. L. The genetic basis of antibody production: a single heavy chain variable region gene encodes all molecules bearing the dominant anti-arsonate idiotype in the strain A mouse. Eur J Immunol. 1983 Feb;13(2):123–132. doi: 10.1002/eji.1830130207. [DOI] [PubMed] [Google Scholar]
  23. Sims J., Rabbitts T. H., Estess P., Slaughter C., Tucker P. W., Capra J. D. Somatic mutation in genes for the variable portion of the immunoglobulin heavy chain. Science. 1982 Apr 16;216(4543):309–311. doi: 10.1126/science.6801765. [DOI] [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