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. 1995 Mar 25;23(6):1060–1067. doi: 10.1093/nar/23.6.1060

Coding sequence composition flanking either signal element alters V(D)J recombination efficiency.

N V Boubnov 1, Z P Wills 1, D T Weaver 1
PMCID: PMC306805  PMID: 7731794

Abstract

Lymphoid V(D)J rearrangement is targeted by recombination signal sequences (RSS) bordering V, D or J exons. We demonstrate that the DNA composition of flanking coding positions, particularly poly(A) or poly(T) stretches at one or both RSS, diminishes V(D)J recombination up to 100-fold. Positionally correct cleavages occur in the inhibited reactions, since the junctions formed show the same frequency of precision as uninhibited reactions. Open/shut cleavage/rejoining is not increased at a normal RSS in substrates containing inhibitory A/T homopolymers versus random sequence at a second RSS. Thus recombinase action at both cleavage sites is severely disrupted by modified coding sequences.

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Selected References

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  1. Bender J., Kleckner N. Tn10 insertion specificity is strongly dependent upon sequences immediately adjacent to the target-site consensus sequence. Proc Natl Acad Sci U S A. 1992 Sep 1;89(17):7996–8000. doi: 10.1073/pnas.89.17.7996. [DOI] [PMC free article] [PubMed] [Google Scholar]
  2. Boubnov N. V., Wills Z. P., Weaver D. T. V(D)J recombination coding junction formation without DNA homology: processing of coding termini. Mol Cell Biol. 1993 Nov;13(11):6957–6968. doi: 10.1128/mcb.13.11.6957. [DOI] [PMC free article] [PubMed] [Google Scholar]
  3. Burkhoff A. M., Tullius T. D. The unusual conformation adopted by the adenine tracts in kinetoplast DNA. Cell. 1987 Mar 27;48(6):935–943. doi: 10.1016/0092-8674(87)90702-1. [DOI] [PubMed] [Google Scholar]
  4. Drew H. R., Travers A. A. DNA structural variations in the E. coli tyrT promoter. Cell. 1984 Jun;37(2):491–502. doi: 10.1016/0092-8674(84)90379-9. [DOI] [PubMed] [Google Scholar]
  5. Gerstein R. M., Lieber M. R. Coding end sequence can markedly affect the initiation of V(D)J recombination. Genes Dev. 1993 Jul;7(7B):1459–1469. doi: 10.1101/gad.7.7b.1459. [DOI] [PubMed] [Google Scholar]
  6. Gilfillan S., Dierich A., Lemeur M., Benoist C., Mathis D. Mice lacking TdT: mature animals with an immature lymphocyte repertoire. Science. 1993 Aug 27;261(5125):1175–1178. doi: 10.1126/science.8356452. [DOI] [PubMed] [Google Scholar]
  7. Hagerman P. J. Sequence-directed curvature of DNA. Annu Rev Biochem. 1990;59:755–781. doi: 10.1146/annurev.bi.59.070190.003543. [DOI] [PubMed] [Google Scholar]
  8. Hendrickson E. A., Liu V. F., Weaver D. T. Strand breaks without DNA rearrangement in V (D)J recombination. Mol Cell Biol. 1991 Jun;11(6):3155–3162. doi: 10.1128/mcb.11.6.3155. [DOI] [PMC free article] [PubMed] [Google Scholar]
  9. Hesse J. E., Lieber M. R., Gellert M., Mizuuchi K. Extrachromosomal DNA substrates in pre-B cells undergo inversion or deletion at immunoglobulin V-(D)-J joining signals. Cell. 1987 Jun 19;49(6):775–783. doi: 10.1016/0092-8674(87)90615-5. [DOI] [PubMed] [Google Scholar]
  10. Hesse J. E., Lieber M. R., Mizuuchi K., Gellert M. V(D)J recombination: a functional definition of the joining signals. Genes Dev. 1989 Jul;3(7):1053–1061. doi: 10.1101/gad.3.7.1053. [DOI] [PubMed] [Google Scholar]
  11. Huisman O., Faelen M., Girard D., Jaffé A., Toussaint A., Rouvière-Yaniv J. Multiple defects in Escherichia coli mutants lacking HU protein. J Bacteriol. 1989 Jul;171(7):3704–3712. doi: 10.1128/jb.171.7.3704-3712.1989. [DOI] [PMC free article] [PubMed] [Google Scholar]
  12. Komori T., Okada A., Stewart V., Alt F. W. Lack of N regions in antigen receptor variable region genes of TdT-deficient lymphocytes. Science. 1993 Aug 27;261(5125):1171–1175. doi: 10.1126/science.8356451. [DOI] [PubMed] [Google Scholar]
  13. Leroy J. L., Charretier E., Kochoyan M., Guéron M. Evidence from base-pair kinetics for two types of adenine tract structures in solution: their relation to DNA curvature. Biochemistry. 1988 Dec 13;27(25):8894–8898. doi: 10.1021/bi00425a004. [DOI] [PubMed] [Google Scholar]
  14. Lewis S. M., Hesse J. E. Cutting and closing without recombination in V(D)J joining. EMBO J. 1991 Dec;10(12):3631–3639. doi: 10.1002/j.1460-2075.1991.tb04929.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  15. Nadeau J. G., Crothers D. M. Structural basis for DNA bending. Proc Natl Acad Sci U S A. 1989 Apr;86(8):2622–2626. doi: 10.1073/pnas.86.8.2622. [DOI] [PMC free article] [PubMed] [Google Scholar]
  16. Oltz E. M., Alt F. W., Lin W. C., Chen J., Taccioli G., Desiderio S., Rathbun G. A V(D)J recombinase-inducible B-cell line: role of transcriptional enhancer elements in directing V(D)J recombination. Mol Cell Biol. 1993 Oct;13(10):6223–6230. doi: 10.1128/mcb.13.10.6223. [DOI] [PMC free article] [PubMed] [Google Scholar]
  17. Pergola F., Zdzienicka M. Z., Lieber M. R. V(D)J recombination in mammalian cell mutants defective in DNA double-strand break repair. Mol Cell Biol. 1993 Jun;13(6):3464–3471. doi: 10.1128/mcb.13.6.3464. [DOI] [PMC free article] [PubMed] [Google Scholar]
  18. Petrini J. H., Donovan J. W., Dimare C., Weaver D. T. Normal V(D)J coding junction formation in DNA ligase I deficiency syndromes. J Immunol. 1994 Jan 1;152(1):176–183. [PubMed] [Google Scholar]
  19. Roth D. B., Zhu C., Gellert M. Characterization of broken DNA molecules associated with V(D)J recombination. Proc Natl Acad Sci U S A. 1993 Nov 15;90(22):10788–10792. doi: 10.1073/pnas.90.22.10788. [DOI] [PMC free article] [PubMed] [Google Scholar]
  20. Schlissel M., Constantinescu A., Morrow T., Baxter M., Peng A. Double-strand signal sequence breaks in V(D)J recombination are blunt, 5'-phosphorylated, RAG-dependent, and cell cycle regulated. Genes Dev. 1993 Dec;7(12B):2520–2532. doi: 10.1101/gad.7.12b.2520. [DOI] [PubMed] [Google Scholar]
  21. Taccioli G. E., Cheng H. L., Varghese A. J., Whitmore G., Alt F. W. A DNA repair defect in Chinese hamster ovary cells affects V(D)J recombination similarly to the murine scid mutation. J Biol Chem. 1994 Mar 11;269(10):7439–7442. [PubMed] [Google Scholar]
  22. Tonegawa S. Somatic generation of antibody diversity. Nature. 1983 Apr 14;302(5909):575–581. doi: 10.1038/302575a0. [DOI] [PubMed] [Google Scholar]
  23. Weaver D. T. V(D)J recombination and double-strand break repair. Adv Immunol. 1995;58:29–85. doi: 10.1016/s0065-2776(08)60619-7. [DOI] [PubMed] [Google Scholar]
  24. Wu Z., Chaconas G. Flanking host sequences can exert an inhibitory effect on the cleavage step of the in vitro mu DNA strand transfer reaction. J Biol Chem. 1992 May 15;267(14):9552–9558. [PubMed] [Google Scholar]

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