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. 1983 Nov;80(22):6967–6971. doi: 10.1073/pnas.80.22.6967

Hybrid gamma 2b-gamma 2a genes expressed in myeloma variants: evidence for homologous recombination.

S A Tilley, L A Eckhardt, K B Marcu, B K Birshtein
PMCID: PMC390107  PMID: 6316338

Abstract

The expressed immunoglobulin heavy chain genes of five gamma 2b-gamma 2a hybrid chain-producing variants of the mouse myeloma MPC-11 (gamma 2b, kappa) have been characterized by genomic Southern blot analysis. Results show that a hybrid gamma 2b-gamma 2a gene was formed in each variant by recombination between the expressed gamma 2b gene of MPC-11 and a gamma 2a gene. The recombination sites are within regions of marked homology between gamma 2b and gamma 2a genes: at least three and probably four variants show gamma 2b-gamma 2a recombination within the heavy chain constant region 2 (CH2) domain, while the fifth has its recombination site between the penultimate nucleotide of CH1 and the eighth nucleotide of the hinge. An unexpected finding is that the hybrid heavy chain-producing variants fall into two subgroups based on their use of different gamma 2a gene forms in hybrid gene formation. This result leads to the speculation that either a tandem gamma 2a gene duplication was present in MPC-11 prior to variant generation or mitotic recombination between chromosomes occurred in the generation of one variant subgroup. The similarity of hybrid gene formation in MPC-11 variants to that apparently responsible for concerted evolution within multigene families and hybrid protein expression in various individuals is noted, and the possible relationship between hybrid gene formation and the heavy chain class switch is discussed.

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Selected References

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

  1. Arnaud P., Wang A. C., Gianazza E., Wang I. Y., Lasne Y., Creyssel R., Fudenberg H. H. Gamma heavy chain disease protein CHA: immunological and structural studies. Mol Immunol. 1981 May;18(5):379–384. doi: 10.1016/0161-5890(81)90099-7. [DOI] [PubMed] [Google Scholar]
  2. Auffray C., Nageotte R., Chambraud B., Rougeon F. Mouse immunoglobulin genes: a bacterial plasmid containing the entire coding sequence for a pre-gamma 2a heavy chain. Nucleic Acids Res. 1980 Mar 25;8(6):1231–1241. doi: 10.1093/nar/8.6.1231. [DOI] [PMC free article] [PubMed] [Google Scholar]
  3. Birshtein B. K., Campbell R., Diamond B. Effects of immunoglobulin structure on Fc receptor binding: a mouse myeloma variant immunoglobulin with a gamma 2b-gamma 2a hybrid heavy chain having a complete gamma 2a Fc region fails to bind to gamma 2a Fc receptors on mouse macrophages. J Immunol. 1982 Aug;129(2):610–614. [PubMed] [Google Scholar]
  4. Birshtein B. K., Campbell R., Greenberg M. L. A gamma 2b-gamma 2a hybrid immunoglobulin heavy chain produced by a variant of the MPC 11 mouse myeloma cell line. Biochemistry. 1980 Apr 29;19(9):1730–1737. doi: 10.1021/bi00550a002. [DOI] [PubMed] [Google Scholar]
  5. Coffino P., Baumal R., Laskov R., Scharff M. D. Cloning of mouse myeloma cells and detection of rare variants. J Cell Physiol. 1972 Jun;79(3):429–440. doi: 10.1002/jcp.1040790313. [DOI] [PubMed] [Google Scholar]
  6. Dildrop R., Brüggemann M., Radbruch A., Rajewsky K., Beyreuther K. Immunoglobulin V region variants in hybridoma cells. II. Recombination between V genes. EMBO J. 1982;1(5):635–640. doi: 10.1002/j.1460-2075.1982.tb01220.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  7. Eckhardt L. A., Tilley S. A., Lang R. B., Marcu K. B., Birshtein B. K. DNA rearrangements in MPC-11 immunoglobulin heavy chain class-switch variants. Proc Natl Acad Sci U S A. 1982 May;79(9):3006–3010. doi: 10.1073/pnas.79.9.3006. [DOI] [PMC free article] [PubMed] [Google Scholar]
  8. Francus T., Dharmgrongartama B., Campbell R., Scharff M. D., Birshtein B. K. IgG2a-producing variants of an IgG2b-producing mouse myeloma cell line. J Exp Med. 1978 Jun 1;147(6):1535–1550. doi: 10.1084/jem.147.6.1535. [DOI] [PMC free article] [PubMed] [Google Scholar]
  9. Honjo T., Nakai S., Nishida Y., Kataoka T., Yamawaki-Kataoka Y., Takahashi N., Obata M., Shimizu A., Yaoita Y., Nikaido T. Rearrangements of immunoglobulin genes during differentiation and evolution. Immunol Rev. 1981;59:33–67. doi: 10.1111/j.1600-065x.1981.tb00455.x. [DOI] [PubMed] [Google Scholar]
  10. Hurwitz J. L., Coleclough C., Cebra J. J. CH gene rearrangements in IgM-bearing B cells and in the normal splenic DNA component of hybridomas making different isotypes of antibody. Cell. 1980 Nov;22(2 Pt 2):349–359. doi: 10.1016/0092-8674(80)90345-1. [DOI] [PubMed] [Google Scholar]
  11. Kedes L. H. Histone genes and histone messengers. Annu Rev Biochem. 1979;48:837–870. doi: 10.1146/annurev.bi.48.070179.004201. [DOI] [PubMed] [Google Scholar]
  12. Kunkel H. G., Natvig J. B., Joslin F. G. A "Lepore" type of hybrid gamma-globulin. Proc Natl Acad Sci U S A. 1969 Jan;62(1):144–149. doi: 10.1073/pnas.62.1.144. [DOI] [PMC free article] [PubMed] [Google Scholar]
  13. Lang R. B., Stanton L. W., Marcu K. B. On immunoglobulin heavy chain gene switching: two gamma 2b genes are rearranged via switch sequences in MPC-11 cells but only one is expressed. Nucleic Acids Res. 1982 Jan 22;10(2):611–630. doi: 10.1093/nar/10.2.611. [DOI] [PMC free article] [PubMed] [Google Scholar]
  14. Liesegang B., Radbruch A., Rajewsky K. Isolation of myeloma variants with predefined variant surface immunoglobulin by cell sorting. Proc Natl Acad Sci U S A. 1978 Aug;75(8):3901–3905. doi: 10.1073/pnas.75.8.3901. [DOI] [PMC free article] [PubMed] [Google Scholar]
  15. Marcu K. B., Banerji J., Penncavage N. A., Lang R., Arnheim N. 5' flanking region of immunoglobulin heavy chain constant region genes displays length heterogeneity in germlines of inbred mouse strains. Cell. 1980 Nov;22(1 Pt 1):187–196. doi: 10.1016/0092-8674(80)90167-1. [DOI] [PubMed] [Google Scholar]
  16. Morrison S. L. Sequentially derived mutants of the constant region of the heavy chain of murine immunoglobulins. J Immunol. 1979 Aug;123(2):793–800. [PubMed] [Google Scholar]
  17. Natvig J. B., Kunkel H. G. A HYBRID IgG4-IgG2 immunoglobulin. J Immunol. 1974 Apr;112(4):1277–1284. [PubMed] [Google Scholar]
  18. Nottenburg C., Weissman I. L. Cmu gene rearrangement of mouse immunoglobulin genes in normal B cells occurs on both the expressed and nonexpressed chromosomes. Proc Natl Acad Sci U S A. 1981 Jan;78(1):484–488. doi: 10.1073/pnas.78.1.484. [DOI] [PMC free article] [PubMed] [Google Scholar]
  19. Ollo R., Auffray C., Morchamps C., Rougeon F. Comparison of mouse immunoglobulin gamma 2a and gamma 2b chain genes suggests that exons can be exchanged between genes in a multigenic family. Proc Natl Acad Sci U S A. 1981 Apr;78(4):2442–2446. doi: 10.1073/pnas.78.4.2442. [DOI] [PMC free article] [PubMed] [Google Scholar]
  20. Ollo R., Rougeon F. Mouse immunoglobulin allotypes: post-duplication divergence of gamma 2a and gamma 2b chain genes. Nature. 1982 Apr 22;296(5859):761–763. doi: 10.1038/296761a0. [DOI] [PubMed] [Google Scholar]
  21. Pease L. R., Schulze D. H., Pfaffenbach G. M., Nathenson S. G. Spontaneous H-2 mutants provide evidence that a copy mechanism analogous to gene conversion generates polymorphism in the major histocompatibility complex. Proc Natl Acad Sci U S A. 1983 Jan;80(1):242–246. doi: 10.1073/pnas.80.1.242. [DOI] [PMC free article] [PubMed] [Google Scholar]
  22. Plunkett M. L., Coligan J. E., David C. S., Freed J. H. Intragenic recombination in an E beta gene for a murine Ia antigen. J Exp Med. 1982 Mar 1;155(3):937–942. doi: 10.1084/jem.155.3.937. [DOI] [PMC free article] [PubMed] [Google Scholar]
  23. Rose S. M., Cullen S. E. A variant alpha-chain in an Ia molecule from the I-A subregion of the mouse major histocompatibility complex: a possible intragenic recombination. J Immunol. 1981 Oct;127(4):1472–1477. [PubMed] [Google Scholar]
  24. Schreier P. H., Bothwell A. L., Mueller-Hill B., Baltimore D. Multiple differences between the nucleic acid sequences of the IgG2aa and IgG2ab alleles of the mouse. Proc Natl Acad Sci U S A. 1981 Jul;78(7):4495–4499. doi: 10.1073/pnas.78.7.4495. [DOI] [PMC free article] [PubMed] [Google Scholar]
  25. Shen S. H., Slightom J. L., Smithies O. A history of the human fetal globin gene duplication. Cell. 1981 Oct;26(2 Pt 2):191–203. doi: 10.1016/0092-8674(81)90302-0. [DOI] [PubMed] [Google Scholar]
  26. Slightom J. L., Blechl A. E., Smithies O. Human fetal G gamma- and A gamma-globin genes: complete nucleotide sequences suggest that DNA can be exchanged between these duplicated genes. Cell. 1980 Oct;21(3):627–638. doi: 10.1016/0092-8674(80)90426-2. [DOI] [PubMed] [Google Scholar]
  27. Takahashi N., Ueda S., Obata M., Nikaido T., Nakai S., Honjo T. Structure of human immunoglobulin gamma genes: implications for evolution of a gene family. Cell. 1982 Jun;29(2):671–679. doi: 10.1016/0092-8674(82)90183-0. [DOI] [PubMed] [Google Scholar]
  28. Towbin H., Staehelin T., Gordon J. Electrophoretic transfer of proteins from polyacrylamide gels to nitrocellulose sheets: procedure and some applications. Proc Natl Acad Sci U S A. 1979 Sep;76(9):4350–4354. doi: 10.1073/pnas.76.9.4350. [DOI] [PMC free article] [PubMed] [Google Scholar]
  29. Tsuzukida Y., Wang C. C., Putnam F. W. Structure of the A2m(1) allotype of human IgA--a recombinant molecule. Proc Natl Acad Sci U S A. 1979 Mar;76(3):1104–1108. doi: 10.1073/pnas.76.3.1104. [DOI] [PMC free article] [PubMed] [Google Scholar]
  30. Tucker P. W., Marcu K. B., Slightom J. L., Blattner F. R. Structure of the constant and 3' untranslated regions of the murine gamma 2b heavy chain messenger RNA. Science. 1979 Dec 14;206(4424):1299–1303. doi: 10.1126/science.117548. [DOI] [PubMed] [Google Scholar]
  31. Warner N. L., Herzenberg L. A., Goldstein G. Immunoglobulin isoantigens (allotypes) in the mouse. II. Allotypic analysis of three gammaG2-myeloma proteins from (NZB x BALB/c)F1 hybrids and of normal gammaG2-globulins. J Exp Med. 1966 Apr 1;123(4):707–721. doi: 10.1084/jem.123.4.707. [DOI] [PMC free article] [PubMed] [Google Scholar]
  32. Weatherall D. J., Clegg J. B. Recent developments in the molecular genetics of human hemoglobin. Cell. 1979 Mar;16(3):467–479. doi: 10.1016/0092-8674(79)90022-9. [DOI] [PubMed] [Google Scholar]
  33. Yamawaki-Kataoka Y., Miyata T., Honjo T. The complete nucleotide sequence of mouse immunoglobin gamma 2a gene and evolution of heavy chain genes: further evidence for intervening sequence-mediated domain transfer. Nucleic Acids Res. 1981 Mar 25;9(6):1365–1381. doi: 10.1093/nar/9.6.1365. [DOI] [PMC free article] [PubMed] [Google Scholar]
  34. Zimmer E. A., Martin S. L., Beverley S. M., Kan Y. W., Wilson A. C. Rapid duplication and loss of genes coding for the alpha chains of hemoglobin. Proc Natl Acad Sci U S A. 1980 Apr;77(4):2158–2162. doi: 10.1073/pnas.77.4.2158. [DOI] [PMC free article] [PubMed] [Google Scholar]

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