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. 1990 Apr;87(8):3072–3076. doi: 10.1073/pnas.87.8.3072

Somatic diversification of chicken immunoglobulin light chains by point mutations.

R Parvari 1, E Ziv 1, F Lantner 1, D Heller 1, I Schechter 1
PMCID: PMC53836  PMID: 2109325

Abstract

The light-chain locus of chicken has 1 functional V lambda 1 gene, 1 J gene, and 25 pseudo-V lambda-genes (where V = variable and J = joining). A major problem is which somatic mechanisms expand this extremely limited germ-line information to generate many different antibodies. Weill's group [Reynaud, C. A., Anquez, V., Grimal, H. & Weill, J. C. (1987) Cell 48, 379-388] has shown that the pseudo-V lambda-genes diversify the rearranged V lambda 1 by gene conversion. Here we demonstrate that chicken light chains are further diversified by somatic point mutations and by V lambda 1-J flexible joining. Somatic point mutations were identified in the J and 3' noncoding DNA of rearranged light-chain genes of chicken. These regions were analyzed because point mutations in V lambda 1 are obscured by gene conversion; the J and 3' noncoding DNA are presented in one copy per haploid genome and are not subject to gene conversion. In rodents point mutations occur as frequently in the V-J coding regions as in the adjacent flanking DNA. Therefore, we conclude that somatic point mutations diversify the V lambda 1 of chicken. The frequency (0-1%) and distribution of the mutations (decreasing in number with increased distance from the V lambda 1 segment) in chicken were as observed in rodents. Sequence variability at the V lambda 1-J junctions could be attributed to imprecise joining of the V lambda 1 and J genes. The modification by gene conversion of rearranged V lambda 1 genes in the bursa was similar in chicken aged 3 months (9.5%) or 3 weeks (9.1%)--i.e., gene conversion that generates the preimmune repertoire in the bursa seems to level off around 3 weeks of age. This preimmune repertoire can be further diversified by somatic point mutations that presumably lead to the formation of antibodies with increased affinity. A segment with structural features of a matrix association region [(A + T)-rich and four topoisomerase II binding sites] was identified in the middle of the J-C lambda intron (where C = constant).

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Selected References

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  1. Bothwell A. L., Paskind M., Reth M., Imanishi-Kari T., Rajewsky K., Baltimore D. Heavy chain variable region contribution to the NPb family of antibodies: somatic mutation evident in a gamma 2a variable region. Cell. 1981 Jun;24(3):625–637. doi: 10.1016/0092-8674(81)90089-1. [DOI] [PubMed] [Google Scholar]
  2. Boyd R. L., Ward H. A. Lymphoid antigenic determinants of the chicken: ontogeny of bursa-dependent lymphoid tissue. Dev Comp Immunol. 1984 Winter;8(1):149–167. doi: 10.1016/0145-305x(84)90019-3. [DOI] [PubMed] [Google Scholar]
  3. Breiner A. V., Brandt C. R., Milcarek C., Sweet R. W., Ziv E., Burstein Y., Schechter I. Somatic DNA rearrangement generates functional rat immunoglobulin kappa chain genes: the J kappa gene cluster is longer in rat than in mouse. Gene. 1982 May;18(2):165–174. doi: 10.1016/0378-1119(82)90114-7. [DOI] [PubMed] [Google Scholar]
  4. COOPER M. D., PETERSON R. D., GOOD R. A. DELINEATION OF THE THYMIC AND BURSAL LYMPHOID SYSTEMS IN THE CHICKEN. Nature. 1965 Jan 9;205:143–146. doi: 10.1038/205143a0. [DOI] [PubMed] [Google Scholar]
  5. Cockerill P. N., Garrard W. T. Chromosomal loop anchorage of the kappa immunoglobulin gene occurs next to the enhancer in a region containing topoisomerase II sites. Cell. 1986 Jan 31;44(2):273–282. doi: 10.1016/0092-8674(86)90761-0. [DOI] [PubMed] [Google Scholar]
  6. French V. I., Stark J. M., White R. G. The influence of adjuvants on the immunological response of the chicken. II. Effects of Freund's complete adjuvant on later antibody production after a single injection of immunogen. Immunology. 1970 May;18(5):645–655. [PMC free article] [PubMed] [Google Scholar]
  7. 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]
  8. 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]
  9. Hozumi N., Wu G. E., Murialdo H., Roberts L., Vetter D., Fife W. L., Whiteley M., Sadowski P. RNA splicing mutation in an aberrantly rearranged immunoglobulin lambda I gene. Proc Natl Acad Sci U S A. 1981 Nov;78(11):7019–7023. doi: 10.1073/pnas.78.11.7019. [DOI] [PMC free article] [PubMed] [Google Scholar]
  10. Jarman A. P., Higgs D. R. Nuclear scaffold attachment sites in the human globin gene complexes. EMBO J. 1988 Nov;7(11):3337–3344. doi: 10.1002/j.1460-2075.1988.tb03205.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  11. Kim S., Davis M., Sinn E., Patten P., Hood L. Antibody diversity: somatic hypermutation of rearranged VH genes. Cell. 1981 Dec;27(3 Pt 2):573–581. doi: 10.1016/0092-8674(81)90399-8. [DOI] [PubMed] [Google Scholar]
  12. Kincade P. W., Cooper M. D. Development and distribution of immunoglobulin-containing cells in the chicken. An immunofluorescent analysis using purified antibodies to mu, gamma and light chains. J Immunol. 1971 Feb;106(2):371–382. [PubMed] [Google Scholar]
  13. Lydyard P. M., Grossi C. E., Cooper M. D. Ontogeny of B cells in the chicken. I. Sequential development of clonal diversity in the bursa. J Exp Med. 1976 Jul 1;144(1):79–97. doi: 10.1084/jem.144.1.79. [DOI] [PMC free article] [PubMed] [Google Scholar]
  14. Max E. E., Seidman J. G., Miller H., Leder P. Variation in the crossover point of kappa immunoglobulin gene V-J recombination: evidence from a cryptic gene. Cell. 1980 Oct;21(3):793–799. doi: 10.1016/0092-8674(80)90442-0. [DOI] [PubMed] [Google Scholar]
  15. McCormack W. T., Tjoelker L. W., Carlson L. M., Petryniak B., Barth C. F., Humphries E. H., Thompson C. B. Chicken IgL gene rearrangement involves deletion of a circular episome and addition of single nonrandom nucleotides to both coding segments. Cell. 1989 Mar 10;56(5):785–791. doi: 10.1016/0092-8674(89)90683-1. [DOI] [PubMed] [Google Scholar]
  16. Parvari R., Avivi A., Lentner F., Ziv E., Tel-Or S., Burstein Y., Schechter I. Chicken immunoglobulin gamma-heavy chains: limited VH gene repertoire, combinatorial diversification by D gene segments and evolution of the heavy chain locus. EMBO J. 1988 Mar;7(3):739–744. doi: 10.1002/j.1460-2075.1988.tb02870.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  17. Parvari R., Ziv E., Lentner F., Tel-Or S., Burstein Y., Schechter I. Analyses of chicken immunoglobulin light chain cDNA clones indicate a few germline V lambda genes and allotypes of the C lambda locus. EMBO J. 1987 Jan;6(1):97–102. doi: 10.1002/j.1460-2075.1987.tb04724.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  18. Pink J. R. Counting components of the chicken's B cell system. Immunol Rev. 1986 Jun;91:115–128. doi: 10.1111/j.1600-065x.1986.tb01486.x. [DOI] [PubMed] [Google Scholar]
  19. Rajewsky K., Förster I., Cumano A. Evolutionary and somatic selection of the antibody repertoire in the mouse. Science. 1987 Nov 20;238(4830):1088–1094. doi: 10.1126/science.3317826. [DOI] [PubMed] [Google Scholar]
  20. Reynaud C. A., Anquez V., Dahan A., Weill J. C. A single rearrangement event generates most of the chicken immunoglobulin light chain diversity. Cell. 1985 Feb;40(2):283–291. doi: 10.1016/0092-8674(85)90142-4. [DOI] [PubMed] [Google Scholar]
  21. Reynaud C. A., Anquez V., Grimal H., Weill J. C. A hyperconversion mechanism generates the chicken light chain preimmune repertoire. Cell. 1987 Feb 13;48(3):379–388. doi: 10.1016/0092-8674(87)90189-9. [DOI] [PubMed] [Google Scholar]
  22. Sander M., Hsieh T. S. Drosophila topoisomerase II double-strand DNA cleavage: analysis of DNA sequence homology at the cleavage site. Nucleic Acids Res. 1985 Feb 25;13(4):1057–1072. doi: 10.1093/nar/13.4.1057. [DOI] [PMC free article] [PubMed] [Google Scholar]
  23. 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]
  24. Thompson C. B., Neiman P. E. Somatic diversification of the chicken immunoglobulin light chain gene is limited to the rearranged variable gene segment. Cell. 1987 Feb 13;48(3):369–378. doi: 10.1016/0092-8674(87)90188-7. [DOI] [PubMed] [Google Scholar]
  25. Weigert M., Perry R., Kelley D., Hunkapiller T., Schilling J., Hood L. The joining of V and J gene segments creates antibody diversity. Nature. 1980 Jan 31;283(5746):497–499. doi: 10.1038/283497a0. [DOI] [PubMed] [Google Scholar]
  26. Weill J. C., Reynaud C. A., Lassila O., Pink J. R. Rearrangement of chicken immunoglobulin genes is not an ongoing process in the embryonic bursa of Fabricius. Proc Natl Acad Sci U S A. 1986 May;83(10):3336–3340. doi: 10.1073/pnas.83.10.3336. [DOI] [PMC free article] [PubMed] [Google Scholar]
  27. Yamaga K., Benedict A. A. Class, amounts, and affinities of anti-dinitrophenyl antibodies in chickens. II. Production of a restricted population of high affinity 7S antibodies by injection of antigen emulsified in adjuvant. J Immunol. 1975 Sep;115(3):759–764. [PubMed] [Google Scholar]

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