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. 1993 Jun;13(6):3611–3622. doi: 10.1128/mcb.13.6.3611

NF-HB (BSAP) is a repressor of the murine immunoglobulin heavy-chain 3' alpha enhancer at early stages of B-cell differentiation.

M Singh 1, B K Birshtein 1
PMCID: PMC359830  PMID: 8497273

Abstract

We have identified a nuclear factor expressed in pro-B-, pre-B-, and B-cell lines that binds to two sites within the murine immunoglobulin heavy-chain (IgH) 3' alpha enhancer (3' alpha E). These sites were defined by oligonucleotide competition in an electrophoretic mobility shift assay (EMSA) and methylation interference footprinting. The 3' alpha E-binding factor is indistinguishable from NF-HB (B-lineage-specific nuclear factor that binds to the IgH gene) and the B-lineage-specific transcription factor BSAP by several criteria, including similar cell type distribution of binding activity, cross-competition of binding sites in EMSA, similar protein size as demonstrated by UV cross-linking, and sequence identity of one of the 3' alpha E-binding sites with a BSAP-binding site within the promoter of the sea urchin late histone gene H2A-2.2. These observations indicate that 3' alpha E is one of the mammalian targets for NF-HB (BSAP). Transient-transfection assays with chloramphenicol acetyltransferase gene constructs containing 3' alpha E and mutant 3' alpha E, in which one of the NF-HB binding sites was inactivated by site-specific mutagenesis, showed ca. five- to sixfold-enhanced activity of mutated 3' alpha E over parental 3' alpha E in B-cell lines (NF-HB+), while no significant difference was observed in plasmacytoma cells (NF-HB-). We conclude from these observations that NF-HB (BSAP) acts as a repressor of the mouse IgH 3' alpha E.

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Selected References

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

  1. Adams B., Dörfler P., Aguzzi A., Kozmik Z., Urbánek P., Maurer-Fogy I., Busslinger M. Pax-5 encodes the transcription factor BSAP and is expressed in B lymphocytes, the developing CNS, and adult testis. Genes Dev. 1992 Sep;6(9):1589–1607. doi: 10.1101/gad.6.9.1589. [DOI] [PubMed] [Google Scholar]
  2. Aguilera R. J., Hope T. J., Sakano H. Characterization of immunoglobulin enhancer deletions in murine plasmacytomas. EMBO J. 1985 Dec 30;4(13B):3689–3693. doi: 10.1002/j.1460-2075.1985.tb04136.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  3. Balling R., Deutsch U., Gruss P. undulated, a mutation affecting the development of the mouse skeleton, has a point mutation in the paired box of Pax 1. Cell. 1988 Nov 4;55(3):531–535. doi: 10.1016/0092-8674(88)90039-6. [DOI] [PubMed] [Google Scholar]
  4. Banerji J., Olson L., Schaffner W. A lymphocyte-specific cellular enhancer is located downstream of the joining region in immunoglobulin heavy chain genes. Cell. 1983 Jul;33(3):729–740. doi: 10.1016/0092-8674(83)90015-6. [DOI] [PubMed] [Google Scholar]
  5. Barberis A., Superti-Furga G., Vitelli L., Kemler I., Busslinger M. Developmental and tissue-specific regulation of a novel transcription factor of the sea urchin. Genes Dev. 1989 May;3(5):663–675. doi: 10.1101/gad.3.5.663. [DOI] [PubMed] [Google Scholar]
  6. Barberis A., Widenhorn K., Vitelli L., Busslinger M. A novel B-cell lineage-specific transcription factor present at early but not late stages of differentiation. Genes Dev. 1990 May;4(5):849–859. doi: 10.1101/gad.4.5.849. [DOI] [PubMed] [Google Scholar]
  7. Boothby M., Gravallese E., Liou H. C., Glimcher L. H. A DNA binding protein regulated by IL-4 and by differentiation in B cells. Science. 1988 Dec 16;242(4885):1559–1562. doi: 10.1126/science.3144043. [DOI] [PubMed] [Google Scholar]
  8. Bopp D., Burri M., Baumgartner S., Frigerio G., Noll M. Conservation of a large protein domain in the segmentation gene paired and in functionally related genes of Drosophila. Cell. 1986 Dec 26;47(6):1033–1040. doi: 10.1016/0092-8674(86)90818-4. [DOI] [PubMed] [Google Scholar]
  9. Chang L. S., Shi Y., Shenk T. Adeno-associated virus P5 promoter contains an adenovirus E1A-inducible element and a binding site for the major late transcription factor. J Virol. 1989 Aug;63(8):3479–3488. doi: 10.1128/jvi.63.8.3479-3488.1989. [DOI] [PMC free article] [PubMed] [Google Scholar]
  10. Dariavach P., Williams G. T., Campbell K., Pettersson S., Neuberger M. S. The mouse IgH 3'-enhancer. Eur J Immunol. 1991 Jun;21(6):1499–1504. doi: 10.1002/eji.1830210625. [DOI] [PubMed] [Google Scholar]
  11. Eckhardt L. A., Birshtein B. K. Independent immunoglobulin class-switch events occurring in a single myeloma cell line. Mol Cell Biol. 1985 Apr;5(4):856–868. doi: 10.1128/mcb.5.4.856. [DOI] [PMC free article] [PubMed] [Google Scholar]
  12. Ephrussi A., Church G. M., Tonegawa S., Gilbert W. B lineage--specific interactions of an immunoglobulin enhancer with cellular factors in vivo. Science. 1985 Jan 11;227(4683):134–140. doi: 10.1126/science.3917574. [DOI] [PubMed] [Google Scholar]
  13. Gerster T., Matthias P., Thali M., Jiricny J., Schaffner W. Cell type-specificity elements of the immunoglobulin heavy chain gene enhancer. EMBO J. 1987 May;6(5):1323–1330. doi: 10.1002/j.1460-2075.1987.tb02371.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  14. Gerster T., Picard D., Schaffner W. During B-cell differentiation enhancer activity and transcription rate of immunoglobulin heavy chain genes are high before mRNA accumulation. Cell. 1986 Apr 11;45(1):45–52. doi: 10.1016/0092-8674(86)90536-2. [DOI] [PubMed] [Google Scholar]
  15. Giannini S. L., Singh M., Calvo C. F., Ding G., Birshtein B. K. DNA regions flanking the mouse Ig 3' alpha enhancer are differentially methylated and DNAase I hypersensitive during B cell differentiation. J Immunol. 1993 Mar 1;150(5):1772–1780. [PubMed] [Google Scholar]
  16. Gillies S. D., Morrison S. L., Oi V. T., Tonegawa S. A tissue-specific transcription enhancer element is located in the major intron of a rearranged immunoglobulin heavy chain gene. Cell. 1983 Jul;33(3):717–728. doi: 10.1016/0092-8674(83)90014-4. [DOI] [PubMed] [Google Scholar]
  17. Grant P. A., Arulampalam V., Ahrlund-Richter L., Pettersson S. Identification of Ets-like lymphoid specific elements within the immunoglobulin heavy chain 3' enhancer. Nucleic Acids Res. 1992 Sep 11;20(17):4401–4408. doi: 10.1093/nar/20.17.4401. [DOI] [PMC free article] [PubMed] [Google Scholar]
  18. Han K., Levine M. S., Manley J. L. Synergistic activation and repression of transcription by Drosophila homeobox proteins. Cell. 1989 Feb 24;56(4):573–583. doi: 10.1016/0092-8674(89)90580-1. [DOI] [PubMed] [Google Scholar]
  19. Hariharan N., Kelley D. E., Perry R. P. Equipotent mouse ribosomal protein promoters have a similar architecture that includes internal sequence elements. Genes Dev. 1989 Nov;3(11):1789–1800. doi: 10.1101/gad.3.11.1789. [DOI] [PubMed] [Google Scholar]
  20. Hollon T., Yoshimura F. K. Variation in enzymatic transient gene expression assays. Anal Biochem. 1989 Nov 1;182(2):411–418. doi: 10.1016/0003-2697(89)90616-7. [DOI] [PubMed] [Google Scholar]
  21. Klein S., Sablitzky F., Radbruch A. Deletion of the IgH enhancer does not reduce immunoglobulin heavy chain production of a hybridoma IgD class switch variant. EMBO J. 1984 Nov;3(11):2473–2476. doi: 10.1002/j.1460-2075.1984.tb02158.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  22. Kozmik Z., Wang S., Dörfler P., Adams B., Busslinger M. The promoter of the CD19 gene is a target for the B-cell-specific transcription factor BSAP. Mol Cell Biol. 1992 Jun;12(6):2662–2672. doi: 10.1128/mcb.12.6.2662. [DOI] [PMC free article] [PubMed] [Google Scholar]
  23. Laemmli U. K. Cleavage of structural proteins during the assembly of the head of bacteriophage T4. Nature. 1970 Aug 15;227(5259):680–685. doi: 10.1038/227680a0. [DOI] [PubMed] [Google Scholar]
  24. Laurenson P., Rine J. Silencers, silencing, and heritable transcriptional states. Microbiol Rev. 1992 Dec;56(4):543–560. doi: 10.1128/mr.56.4.543-560.1992. [DOI] [PMC free article] [PubMed] [Google Scholar]
  25. Liao F., Giannini S. L., Birshtein B. K. A nuclear DNA-binding protein expressed during early stages of B cell differentiation interacts with diverse segments within and 3' of the Ig H chain gene cluster. J Immunol. 1992 May 1;148(9):2909–2917. [PubMed] [Google Scholar]
  26. Libermann T. A., Lenardo M., Baltimore D. Involvement of a second lymphoid-specific enhancer element in the regulation of immunoglobulin heavy-chain gene expression. Mol Cell Biol. 1990 Jun;10(6):3155–3162. doi: 10.1128/mcb.10.6.3155. [DOI] [PMC free article] [PubMed] [Google Scholar]
  27. Lobell R. B., Schleif R. F. AraC-DNA looping: orientation and distance-dependent loop breaking by the cyclic AMP receptor protein. J Mol Biol. 1991 Mar 5;218(1):45–54. doi: 10.1016/0022-2836(91)90872-4. [DOI] [PubMed] [Google Scholar]
  28. Matthias P., Baltimore D. The immunoglobulin heavy chain locus contains another B-cell-specific 3' enhancer close to the alpha constant region. Mol Cell Biol. 1993 Mar;13(3):1547–1553. doi: 10.1128/mcb.13.3.1547. [DOI] [PMC free article] [PubMed] [Google Scholar]
  29. Maxam A. M., Gilbert W. Sequencing end-labeled DNA with base-specific chemical cleavages. Methods Enzymol. 1980;65(1):499–560. doi: 10.1016/s0076-6879(80)65059-9. [DOI] [PubMed] [Google Scholar]
  30. Mitchell P. J., Wang C., Tjian R. Positive and negative regulation of transcription in vitro: enhancer-binding protein AP-2 is inhibited by SV40 T antigen. Cell. 1987 Sep 11;50(6):847–861. doi: 10.1016/0092-8674(87)90512-5. [DOI] [PubMed] [Google Scholar]
  31. Nelsen B., Kadesch T., Sen R. Complex regulation of the immunoglobulin mu heavy-chain gene enhancer: microB, a new determinant of enhancer function. Mol Cell Biol. 1990 Jun;10(6):3145–3154. doi: 10.1128/mcb.10.6.3145. [DOI] [PMC free article] [PubMed] [Google Scholar]
  32. Okabe T., Watanabe T., Kudo A. A pre-B- and B cell-specific DNA-binding protein, EBB-1, which binds to the promoter of the VpreB1 gene. Eur J Immunol. 1992 Jan;22(1):37–43. doi: 10.1002/eji.1830220107. [DOI] [PubMed] [Google Scholar]
  33. Park K., Atchison M. L. Isolation of a candidate repressor/activator, NF-E1 (YY-1, delta), that binds to the immunoglobulin kappa 3' enhancer and the immunoglobulin heavy-chain mu E1 site. Proc Natl Acad Sci U S A. 1991 Nov 1;88(21):9804–9808. doi: 10.1073/pnas.88.21.9804. [DOI] [PMC free article] [PubMed] [Google Scholar]
  34. Pettersson S., Cook G. P., Brüggemann M., Williams G. T., Neuberger M. S. A second B cell-specific enhancer 3' of the immunoglobulin heavy-chain locus. Nature. 1990 Mar 8;344(6262):165–168. doi: 10.1038/344165a0. [DOI] [PubMed] [Google Scholar]
  35. Schreiber E., Matthias P., Müller M. M., Schaffner W. Rapid detection of octamer binding proteins with 'mini-extracts', prepared from a small number of cells. Nucleic Acids Res. 1989 Aug 11;17(15):6419–6419. doi: 10.1093/nar/17.15.6419. [DOI] [PMC free article] [PubMed] [Google Scholar]
  36. Singh H., Sen R., Baltimore D., Sharp P. A. A nuclear factor that binds to a conserved sequence motif in transcriptional control elements of immunoglobulin genes. Nature. 1986 Jan 9;319(6049):154–158. doi: 10.1038/319154a0. [DOI] [PubMed] [Google Scholar]
  37. Wabl M. R., Burrows P. D. Expression of immunoglobulin heavy chain at a high level in the absence of a proposed immunoglobulin enhancer element in cis. Proc Natl Acad Sci U S A. 1984 Apr;81(8):2452–2455. doi: 10.1073/pnas.81.8.2452. [DOI] [PMC free article] [PubMed] [Google Scholar]
  38. Walther C., Guenet J. L., Simon D., Deutsch U., Jostes B., Goulding M. D., Plachov D., Balling R., Gruss P. Pax: a murine multigene family of paired box-containing genes. Genomics. 1991 Oct;11(2):424–434. doi: 10.1016/0888-7543(91)90151-4. [DOI] [PubMed] [Google Scholar]
  39. Waters S. H., Saikh K. U., Stavnezer J. A B-cell-specific nuclear protein that binds to DNA sites 5' to immunoglobulin S alpha tandem repeats is regulated during differentiation. Mol Cell Biol. 1989 Dec;9(12):5594–5601. doi: 10.1128/mcb.9.12.5594. [DOI] [PMC free article] [PubMed] [Google Scholar]
  40. Wirth T., Staudt L., Baltimore D. An octamer oligonucleotide upstream of a TATA motif is sufficient for lymphoid-specific promoter activity. Nature. 1987 Sep 10;329(6135):174–178. doi: 10.1038/329174a0. [DOI] [PubMed] [Google Scholar]
  41. Wu C., Wilson S., Walker B., Dawid I., Paisley T., Zimarino V., Ueda H. Purification and properties of Drosophila heat shock activator protein. Science. 1987 Nov 27;238(4831):1247–1253. doi: 10.1126/science.3685975. [DOI] [PubMed] [Google Scholar]

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