Abstract
We have transferred a pSV2neo vector containing the wild-type constant region of the immunoglobulin mu gene (C mu) into the mutant hybridoma igm482, which bears a 2-bp deletion in the third constant-region exon of its haploid chromosomal mu gene (C mu 3). Independent igm482 transformants contain the wild-type immunoglobulin C mu region stably integrated in ectopic chromosomal positions. We report here that the wild-type immunoglobulin C mu region can function as the donor sequence in a gene conversion event which corrects the 2-bp deletion in the mutant igm482 chromosomal C mu 3 exon. The homologous recombination event restores normal immunoglobulin M production in the mutant cell.
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Selected References
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- 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]
- Alt F. W., Blackwell T. K., Yancopoulos G. D. Development of the primary antibody repertoire. Science. 1987 Nov 20;238(4830):1079–1087. doi: 10.1126/science.3317825. [DOI] [PubMed] [Google Scholar]
- Baker M. D. High-frequency homologous recombination between duplicate chromosomal immunoglobulin mu heavy-chain constant regions. Mol Cell Biol. 1989 Dec;9(12):5500–5507. doi: 10.1128/mcb.9.12.5500. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Baker M. D., Pennell N., Bosnoyan L., Shulman M. J. Homologous recombination can restore normal immunoglobulin production in a mutant hybridoma cell line. Proc Natl Acad Sci U S A. 1988 Sep;85(17):6432–6436. doi: 10.1073/pnas.85.17.6432. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Baltimore D. Gene conversion: some implications for immunoglobulin genes. Cell. 1981 Jun;24(3):592–594. doi: 10.1016/0092-8674(81)90082-9. [DOI] [PubMed] [Google Scholar]
- Baumann B., Potash M. J., Köhler G. Consequences of frameshift mutations at the immunoglobulin heavy chain locus of the mouse. EMBO J. 1985 Feb;4(2):351–359. doi: 10.1002/j.1460-2075.1985.tb03636.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Blackwell T. K., Moore M. W., Yancopoulos G. D., Suh H., Lutzker S., Selsing E., Alt F. W. Recombination between immunoglobulin variable region gene segments is enhanced by transcription. Nature. 1986 Dec 11;324(6097):585–589. doi: 10.1038/324585a0. [DOI] [PubMed] [Google Scholar]
- Blaho J. A., Wells R. D. Left-handed Z-DNA and genetic recombination. Prog Nucleic Acid Res Mol Biol. 1989;37:107–126. doi: 10.1016/s0079-6603(08)60696-0. [DOI] [PubMed] [Google Scholar]
- Bollag R. J., Liskay R. M. Conservative intrachromosomal recombination between inverted repeats in mouse cells: association between reciprocal exchange and gene conversion. Genetics. 1988 May;119(1):161–169. doi: 10.1093/genetics/119.1.161. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Borst P., Greaves D. R. Programmed gene rearrangements altering gene expression. Science. 1987 Feb 6;235(4789):658–667. doi: 10.1126/science.3544215. [DOI] [PubMed] [Google Scholar]
- Bullock P., Miller J., Botchan M. Effects of poly[d(pGpT).d(pApC)] and poly[d(pCpG).d(pCpG)] repeats on homologous recombination in somatic cells. Mol Cell Biol. 1986 Nov;6(11):3948–3953. doi: 10.1128/mcb.6.11.3948. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Capecchi M. R. Altering the genome by homologous recombination. Science. 1989 Jun 16;244(4910):1288–1292. doi: 10.1126/science.2660260. [DOI] [PubMed] [Google Scholar]
- Capizzi R. L., Jameson J. W. A table for the estimation of the spontaneous mutation rate of cells in culture. Mutat Res. 1973 Jan;17(1):147–148. doi: 10.1016/0027-5107(73)90265-0. [DOI] [PubMed] [Google Scholar]
- Cohen J. B., Effron K., Rechavi G., Ben-Neriah Y., Zakut R., Givol D. Simple DNA sequences in homologous flanking regions near immunoglobulin VH genes: a role in gene interaction? Nucleic Acids Res. 1982 Jun 11;10(11):3353–3370. doi: 10.1093/nar/10.11.3353. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Egel R. Intergenic conversion and reiterated genes. Nature. 1981 Mar 19;290(5803):191–192. doi: 10.1038/290191a0. [DOI] [PubMed] [Google Scholar]
- Ernst J. F., Stewart J. W., Sherman F. The cyc1-11 mutation in yeast reverts by recombination with a nonallelic gene: composite genes determining the iso-cytochromes c. Proc Natl Acad Sci U S A. 1981 Oct;78(10):6334–6338. doi: 10.1073/pnas.78.10.6334. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Esser C., Radbruch A. Immunoglobulin class switching: molecular and cellular analysis. Annu Rev Immunol. 1990;8:717–735. doi: 10.1146/annurev.iy.08.040190.003441. [DOI] [PubMed] [Google Scholar]
- Felsenfeld G. Chromatin as an essential part of the transcriptional mechanism. Nature. 1992 Jan 16;355(6357):219–224. doi: 10.1038/355219a0. [DOI] [PubMed] [Google Scholar]
- Finger L. R., Harvey R. C., Moore R. C., Showe L. C., Croce C. M. A common mechanism of chromosomal translocation in T- and B-cell neoplasia. Science. 1986 Nov 21;234(4779):982–985. doi: 10.1126/science.3490692. [DOI] [PubMed] [Google Scholar]
- Flavell R. A., Allen H., Burkly L. C., Sherman D. H., Waneck G. L., Widera G. Molecular biology of the H-2 histocompatibility complex. Science. 1986 Jul 25;233(4762):437–443. doi: 10.1126/science.3726537. [DOI] [PubMed] [Google Scholar]
- Gerstein R. M., Frankel W. N., Hsieh C. L., Durdik J. M., Rath S., Coffin J. M., Nisonoff A., Selsing E. Isotype switching of an immunoglobulin heavy chain transgene occurs by DNA recombination between different chromosomes. Cell. 1990 Nov 2;63(3):537–548. doi: 10.1016/0092-8674(90)90450-s. [DOI] [PubMed] [Google Scholar]
- Goldberg G. I., Vanin E. F., Zrolka A. M., Blattner F. R. Sequence of the gene for the constant region of the mu chain of Balb/c mouse immunoglobulin. Gene. 1981 Oct;15(1):33–42. doi: 10.1016/0378-1119(81)90102-5. [DOI] [PubMed] [Google Scholar]
- Gross-Bellard M., Oudet P., Chambon P. Isolation of high-molecular-weight DNA from mammalian cells. Eur J Biochem. 1973 Jul 2;36(1):32–38. doi: 10.1111/j.1432-1033.1973.tb02881.x. [DOI] [PubMed] [Google Scholar]
- Haluska F. G., Tsujimoto Y., Croce C. M. Oncogene activation by chromosome translocation in human malignancy. Annu Rev Genet. 1987;21:321–345. doi: 10.1146/annurev.ge.21.120187.001541. [DOI] [PubMed] [Google Scholar]
- Jerne N. K., Henry C., Nordin A. A., Fuji H., Koros A. M., Lefkovits I. Plaque forming cells: methodology and theory. Transplant Rev. 1974;18:130–191. doi: 10.1111/j.1600-065x.1974.tb01588.x. [DOI] [PubMed] [Google Scholar]
- Keil R. L., Roeder G. S. Cis-acting, recombination-stimulating activity in a fragment of the ribosomal DNA of S. cerevisiae. Cell. 1984 Dec;39(2 Pt 1):377–386. doi: 10.1016/0092-8674(84)90016-3. [DOI] [PubMed] [Google Scholar]
- Klar A. J., Strathern J. N., Hicks J. B. A position-effect control for gene transposition: state of expression of yeast mating-type genes affects their ability to switch. Cell. 1981 Aug;25(2):517–524. doi: 10.1016/0092-8674(81)90070-2. [DOI] [PubMed] [Google Scholar]
- Köhler G., Potash M. J., Lehrach H., Shulman M. J. Deletions in immunoglobulin mu chains. EMBO J. 1982;1(5):555–563. doi: 10.1002/j.1460-2075.1982.tb01208.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Lennon G. G., Perry R. P. The temporal order of appearance of transcripts from unrearranged and rearranged Ig genes in murine fetal liver. J Immunol. 1990 Mar 1;144(5):1983–1987. [PubMed] [Google Scholar]
- Liskay R. M., Letsou A., Stachelek J. L. Homology requirement for efficient gene conversion between duplicated chromosomal sequences in mammalian cells. Genetics. 1987 Jan;115(1):161–167. doi: 10.1093/genetics/115.1.161. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Liskay R. M., Stachelek J. L. Information transfer between duplicated chromosomal sequences in mammalian cells involves contiguous regions of DNA. Proc Natl Acad Sci U S A. 1986 Mar;83(6):1802–1806. doi: 10.1073/pnas.83.6.1802. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Liskay R. M., Stachelek J. L., Letsou A. Homologous recombination between repeated chromosomal sequences in mouse cells. Cold Spring Harb Symp Quant Biol. 1984;49:183–189. doi: 10.1101/sqb.1984.049.01.021. [DOI] [PubMed] [Google Scholar]
- Maizels N. Might gene conversion be the mechanism of somatic hypermutation of mammalian immunoglobulin genes? Trends Genet. 1989 Jan;5(1):4–8. doi: 10.1016/0168-9525(89)90004-8. [DOI] [PubMed] [Google Scholar]
- McCormack W. T., Tjoelker L. W., Thompson C. B. Avian B-cell development: generation of an immunoglobulin repertoire by gene conversion. Annu Rev Immunol. 1991;9:219–241. doi: 10.1146/annurev.iy.09.040191.001251. [DOI] [PubMed] [Google Scholar]
- Mellor A. L., Weiss E. H., Ramachandran K., Flavell R. A. A potential donor gene for the bm1 gene conversion event in the C57BL mouse. Nature. 1983 Dec 22;306(5945):792–795. doi: 10.1038/306792a0. [DOI] [PubMed] [Google Scholar]
- Morrison S. L., Scharff M. D. Mutational events in mouse myeloma cells. Crit Rev Immunol. 1981 Sep;3(1):1–22. [PubMed] [Google Scholar]
- Nicholls R. D., Fischel-Ghodsian N., Higgs D. R. Recombination at the human alpha-globin gene cluster: sequence features and topological constraints. Cell. 1987 May 8;49(3):369–378. doi: 10.1016/0092-8674(87)90289-3. [DOI] [PubMed] [Google Scholar]
- 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]
- O'Gorman S., Fox D. T., Wahl G. M. Recombinase-mediated gene activation and site-specific integration in mammalian cells. Science. 1991 Mar 15;251(4999):1351–1355. doi: 10.1126/science.1900642. [DOI] [PubMed] [Google Scholar]
- Powers P. A., Smithies O. Short gene conversions in the human fetal globin gene region: a by-product of chromosome pairing during meiosis? Genetics. 1986 Feb;112(2):343–358. doi: 10.1093/genetics/112.2.343. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Richards J. E., Gilliam A. C., Shen A., Tucker P. W., Blattner F. R. Unusual sequences in the murine immunoglobulin mu-delta heavy-chain region. Nature. 1983 Dec 1;306(5942):483–487. doi: 10.1038/306483a0. [DOI] [PubMed] [Google Scholar]
- Rubnitz J., Subramani S. Extrachromosomal and chromosomal gene conversion in mammalian cells. Mol Cell Biol. 1986 May;6(5):1608–1614. doi: 10.1128/mcb.6.5.1608. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Schlissel M. S., Baltimore D. Activation of immunoglobulin kappa gene rearrangement correlates with induction of germline kappa gene transcription. Cell. 1989 Sep 8;58(5):1001–1007. doi: 10.1016/0092-8674(89)90951-3. [DOI] [PubMed] [Google Scholar]
- Shimizu A., Honjo T. Immunoglobulin class switching. Cell. 1984 Apr;36(4):801–803. doi: 10.1016/0092-8674(84)90029-1. [DOI] [PubMed] [Google Scholar]
- Shulman M. J., Heusser C., Filkin C., Köhler G. Mutations affecting the structure and function of immunoglobulin M. Mol Cell Biol. 1982 Sep;2(9):1033–1043. doi: 10.1128/mcb.2.9.1033. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Shulman M. J., Nissen L., Collins C. Homologous recombination in hybridoma cells: dependence on time and fragment length. Mol Cell Biol. 1990 Sep;10(9):4466–4472. doi: 10.1128/mcb.10.9.4466. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Smith A. J., Berg P. Homologous recombination between defective neo genes in mouse 3T6 cells. Cold Spring Harb Symp Quant Biol. 1984;49:171–181. doi: 10.1101/sqb.1984.049.01.020. [DOI] [PubMed] [Google Scholar]
- 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]
- Southern P. J., Berg P. Transformation of mammalian cells to antibiotic resistance with a bacterial gene under control of the SV40 early region promoter. J Mol Appl Genet. 1982;1(4):327–341. [PubMed] [Google Scholar]
- Strathern J. N., Klar A. J., Hicks J. B., Abraham J. A., Ivy J. M., Nasmyth K. A., McGill C. Homothallic switching of yeast mating type cassettes is initiated by a double-stranded cut in the MAT locus. Cell. 1982 Nov;31(1):183–192. doi: 10.1016/0092-8674(82)90418-4. [DOI] [PubMed] [Google Scholar]
- Stringer J. R. Recombination between poly[d(GT).d(CA)] sequences in simian virus 40-infected cultured cells. Mol Cell Biol. 1985 Jun;5(6):1247–1259. doi: 10.1128/mcb.5.6.1247. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Subramani S., Rubnitz J. Recombination events after transient infection and stable integration of DNA into mouse cells. Mol Cell Biol. 1985 Apr;5(4):659–666. doi: 10.1128/mcb.5.4.659. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Taub R. A., Hollis G. F., Hieter P. A., Korsmeyer S., Waldmann T. A., Leder P. Variable amplification of immunoglobulin lambda light-chain genes in human populations. Nature. 1983 Jul 14;304(5922):172–174. doi: 10.1038/304172a0. [DOI] [PubMed] [Google Scholar]
- Thomas B. J., Rothstein R. Elevated recombination rates in transcriptionally active DNA. Cell. 1989 Feb 24;56(4):619–630. doi: 10.1016/0092-8674(89)90584-9. [DOI] [PubMed] [Google Scholar]
- Timsit Y., Vilbois E., Moras D. Base-pairing shift in the major groove of (CA)n tracts by B-DNA crystal structures. Nature. 1991 Nov 14;354(6349):167–170. doi: 10.1038/354167a0. [DOI] [PubMed] [Google Scholar]
- Treco D., Arnheim N. The evolutionarily conserved repetitive sequence d(TG.AC)n promotes reciprocal exchange and generates unusual recombinant tetrads during yeast meiosis. Mol Cell Biol. 1986 Nov;6(11):3934–3947. doi: 10.1128/mcb.6.11.3934. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Trimble W. S., Baker M. D., Boulianne G. L., Murialdo H., Hozumi N., Shulman M. J. Analysis of hybridoma mutants defective in synthesis of immunoglobulin M. Somat Cell Mol Genet. 1986 Sep;12(5):467–477. doi: 10.1007/BF01539918. [DOI] [PubMed] [Google Scholar]
- Tsujimoto Y., Gorham J., Cossman J., Jaffe E., Croce C. M. The t(14;18) chromosome translocations involved in B-cell neoplasms result from mistakes in VDJ joining. Science. 1985 Sep 27;229(4720):1390–1393. doi: 10.1126/science.3929382. [DOI] [PubMed] [Google Scholar]
- Voelkel-Meiman K., Keil R. L., Roeder G. S. Recombination-stimulating sequences in yeast ribosomal DNA correspond to sequences regulating transcription by RNA polymerase I. Cell. 1987 Mar 27;48(6):1071–1079. doi: 10.1016/0092-8674(87)90714-8. [DOI] [PubMed] [Google Scholar]
- Wahls W. P., Wallace L. J., Moore P. D. The Z-DNA motif d(TG)30 promotes reception of information during gene conversion events while stimulating homologous recombination in human cells in culture. Mol Cell Biol. 1990 Feb;10(2):785–793. doi: 10.1128/mcb.10.2.785. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Walsh J. B. Sequence-dependent gene conversion: can duplicated genes diverge fast enough to escape conversion? Genetics. 1987 Nov;117(3):543–557. doi: 10.1093/genetics/117.3.543. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Wittig B., Dorbic T., Rich A. Transcription is associated with Z-DNA formation in metabolically active permeabilized mammalian cell nuclei. Proc Natl Acad Sci U S A. 1991 Mar 15;88(6):2259–2263. doi: 10.1073/pnas.88.6.2259. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Wysocki L. J., Gefter M. L. Gene conversion and the generation of antibody diversity. Annu Rev Biochem. 1989;58:509–531. doi: 10.1146/annurev.bi.58.070189.002453. [DOI] [PubMed] [Google Scholar]
- Yu H., Eckhardt L. A. DNA rearrangement causes a high rate of spontaneous mutation at the immunoglobulin heavy-chain locus of a mouse myeloma cell line. Mol Cell Biol. 1986 Dec;6(12):4228–4235. doi: 10.1128/mcb.6.12.4228. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Zoller M. J., Smith M. Oligonucleotide-directed mutagenesis using M13-derived vectors: an efficient and general procedure for the production of point mutations in any fragment of DNA. Nucleic Acids Res. 1982 Oct 25;10(20):6487–6500. doi: 10.1093/nar/10.20.6487. [DOI] [PMC free article] [PubMed] [Google Scholar]