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. 1997 Oct 15;25(20):3995–4003. doi: 10.1093/nar/25.20.3995

Efficient transcription of an immunoglobulin kappa promoter requires specific sequence elements overlapping with and downstream of the transcriptional start site.

M R Pelletier 1, E N Hatada 1, G Scholz 1, C Scheidereit 1
PMCID: PMC147016  PMID: 9321649

Abstract

The expression of immunoglobulin (Ig) genes depends on tissue-specific elements in the promoter and enhancer regions of light chain and heavy chain genes. In contrast to the complex modular character of Ig enhancers, the promoters appear to be simple, depending primarily on a conserved TATA box and octamer elements. We have analyzed the role of proximal sequences for Igkappa promoter function. Igkappa promoter transcription critically depends on initiator-like sequences and on a downstream element located at +24 to +39 relative to the start site. Replacement of these sequences resulted in strong reduction of promoter activity. In vitro, these elements were found to be more effective in extracts of lymphoid than of non-lymphoid origin. Deletion of the downstream and initiation site regions had a comparable effect on promoter activity to obliteration of the TATA box or octamer element. The downstream sequence was bound by two nuclear proteins, identical to the previously identified Ig-specific C5 and C6 complexes. Whereas C5 is found in HeLa cells and in lymphoid cells, C6 is lymphoid specific. Thus, further specific sequences in addition to the previously characterized elements, the octamer and the TATA box, are required for efficient kappa promoter expression in B lymphocytes.

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Selected References

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  1. Altenburger W., Steinmetz M., Zachau H. G. Functional and non-functional joining in immunoglobulin light chain genes of a mouse myeloma. Nature. 1980 Oct 16;287(5783):603–607. doi: 10.1038/287603a0. [DOI] [PubMed] [Google Scholar]
  2. Arnosti D. N., Merino A., Reinberg D., Schaffner W. Oct-2 facilitates functional preinitiation complex assembly and is continuously required at the promoter for multiple rounds of transcription. EMBO J. 1993 Jan;12(1):157–166. doi: 10.1002/j.1460-2075.1993.tb05641.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  3. Atchison M. L., Delmas V., Perry R. P. A novel upstream element compensates for an ineffectual octamer motif in an immunoglobulin V kappa promoter. EMBO J. 1990 Oct;9(10):3109–3117. doi: 10.1002/j.1460-2075.1990.tb07508.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  4. Ayer D. E., Dynan W. S. Simian virus 40 major late promoter: a novel tripartite structure that includes intragenic sequences. Mol Cell Biol. 1988 May;8(5):2021–2033. doi: 10.1128/mcb.8.5.2021. [DOI] [PMC free article] [PubMed] [Google Scholar]
  5. Burke T. W., Kadonaga J. T. Drosophila TFIID binds to a conserved downstream basal promoter element that is present in many TATA-box-deficient promoters. Genes Dev. 1996 Mar 15;10(6):711–724. doi: 10.1101/gad.10.6.711. [DOI] [PubMed] [Google Scholar]
  6. Clausell A., Tucker P. W. Functional analysis of the V gamma 3 promoter of the murine gamma delta T-cell receptor. Mol Cell Biol. 1994 Jan;14(1):803–814. doi: 10.1128/mcb.14.1.803. [DOI] [PMC free article] [PubMed] [Google Scholar]
  7. Clerc R. G., Corcoran L. M., LeBowitz J. H., Baltimore D., Sharp P. A. The B-cell-specific Oct-2 protein contains POU box- and homeo box-type domains. Genes Dev. 1988 Dec;2(12A):1570–1581. doi: 10.1101/gad.2.12a.1570. [DOI] [PubMed] [Google Scholar]
  8. Cooper C., Johnson D., Roman C., Avitahl N., Tucker P., Calame K. The C/EBP family of transcriptional activators is functionally important for Ig VH promoter activity in vivo and in vitro. J Immunol. 1992 Nov 15;149(10):3225–3231. [PubMed] [Google Scholar]
  9. Corcoran L. M., Karvelas M., Nossal G. J., Ye Z. S., Jacks T., Baltimore D. Oct-2, although not required for early B-cell development, is critical for later B-cell maturation and for postnatal survival. Genes Dev. 1993 Apr;7(4):570–582. doi: 10.1101/gad.7.4.570. [DOI] [PubMed] [Google Scholar]
  10. Dignam J. D., Lebovitz R. M., Roeder R. G. Accurate transcription initiation by RNA polymerase II in a soluble extract from isolated mammalian nuclei. Nucleic Acids Res. 1983 Mar 11;11(5):1475–1489. doi: 10.1093/nar/11.5.1475. [DOI] [PMC free article] [PubMed] [Google Scholar]
  11. Emanuel P. A., Gilmour D. S. Transcription factor TFIID recognizes DNA sequences downstream of the TATA element in the Hsp70 heat shock gene. Proc Natl Acad Sci U S A. 1993 Sep 15;90(18):8449–8453. doi: 10.1073/pnas.90.18.8449. [DOI] [PMC free article] [PubMed] [Google Scholar]
  12. Falkner F. G., Zachau H. G. Correct transcription of an immunoglobulin kappa gene requires an upstream fragment containing conserved sequence elements. Nature. 1984 Jul 5;310(5972):71–74. doi: 10.1038/310071a0. [DOI] [PubMed] [Google Scholar]
  13. Franke S., Scholz G., Scheidereit C. Identification of novel ubiquitous and cell type-specific factors that specifically recognize immunoglobulin heavy chain and kappa light chain promoters. J Biol Chem. 1994 Aug 5;269(31):20075–20082. [PubMed] [Google Scholar]
  14. Gill G. Transcriptional initiation. Taking the initiative. Curr Biol. 1994 Apr 1;4(4):374–376. doi: 10.1016/s0960-9822(00)00084-1. [DOI] [PubMed] [Google Scholar]
  15. Gstaiger M., Knoepfel L., Georgiev O., Schaffner W., Hovens C. M. A B-cell coactivator of octamer-binding transcription factors. Nature. 1995 Jan 26;373(6512):360–362. doi: 10.1038/373360a0. [DOI] [PubMed] [Google Scholar]
  16. Horikoshi M., Hai T., Lin Y. S., Green M. R., Roeder R. G. Transcription factor ATF interacts with the TATA factor to facilitate establishment of a preinitiation complex. Cell. 1988 Sep 23;54(7):1033–1042. doi: 10.1016/0092-8674(88)90118-3. [DOI] [PubMed] [Google Scholar]
  17. Jarrell K. A., Meselson M. Drosophila retrotransposon promoter includes an essential sequence at the initiation site and requires a downstream sequence for full activity. Proc Natl Acad Sci U S A. 1991 Jan 1;88(1):102–104. doi: 10.1073/pnas.88.1.102. [DOI] [PMC free article] [PubMed] [Google Scholar]
  18. Javahery R., Khachi A., Lo K., Zenzie-Gregory B., Smale S. T. DNA sequence requirements for transcriptional initiator activity in mammalian cells. Mol Cell Biol. 1994 Jan;14(1):116–127. doi: 10.1128/mcb.14.1.116. [DOI] [PMC free article] [PubMed] [Google Scholar]
  19. Jenuwein T., Grosschedl R. Complex pattern of immunoglobulin mu gene expression in normal and transgenic mice: nonoverlapping regulatory sequences govern distinct tissue specificities. Genes Dev. 1991 Jun;5(6):932–943. doi: 10.1101/gad.5.6.932. [DOI] [PubMed] [Google Scholar]
  20. Jones K. A., Luciw P. A., Duchange N. Structural arrangements of transcription control domains within the 5'-untranslated leader regions of the HIV-1 and HIV-2 promoters. Genes Dev. 1988 Sep;2(9):1101–1114. doi: 10.1101/gad.2.9.1101. [DOI] [PubMed] [Google Scholar]
  21. Kashanchi F., Duvall J. F., Dittmer J., Mireskandari A., Reid R. L., Gitlin S. D., Brady J. N. Involvement of transcription factor YB-1 in human T-cell lymphotropic virus type I basal gene expression. J Virol. 1994 Jan;68(1):561–565. doi: 10.1128/jvi.68.1.561-565.1994. [DOI] [PMC free article] [PubMed] [Google Scholar]
  22. Kato H., Horikoshi M., Roeder R. G. Repression of HIV-1 transcription by a cellular protein. Science. 1991 Mar 22;251(5000):1476–1479. doi: 10.1126/science.2006421. [DOI] [PubMed] [Google Scholar]
  23. Kim J., Parvin J. D., Shykind B. M., Sharp P. A. A negative cofactor containing Dr1/p19 modulates transcription with TFIIA in a promoter-specific fashion. J Biol Chem. 1996 Aug 2;271(31):18405–18412. doi: 10.1074/jbc.271.31.18405. [DOI] [PubMed] [Google Scholar]
  24. Kim U., Qin X. F., Gong S., Stevens S., Luo Y., Nussenzweig M., Roeder R. G. The B-cell-specific transcription coactivator OCA-B/OBF-1/Bob-1 is essential for normal production of immunoglobulin isotypes. Nature. 1996 Oct 10;383(6600):542–547. doi: 10.1038/383542a0. [DOI] [PubMed] [Google Scholar]
  25. Luo Y., Roeder R. G. Cloning, functional characterization, and mechanism of action of the B-cell-specific transcriptional coactivator OCA-B. Mol Cell Biol. 1995 Aug;15(8):4115–4124. doi: 10.1128/mcb.15.8.4115. [DOI] [PMC free article] [PubMed] [Google Scholar]
  26. Martinez E., Chiang C. M., Ge H., Roeder R. G. TATA-binding protein-associated factor(s) in TFIID function through the initiator to direct basal transcription from a TATA-less class II promoter. EMBO J. 1994 Jul 1;13(13):3115–3126. doi: 10.1002/j.1460-2075.1994.tb06610.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  27. Mizushima-Sugano J., Roeder R. G. Cell-type-specific transcription of an immunoglobulin kappa light chain gene in vitro. Proc Natl Acad Sci U S A. 1986 Nov;83(22):8511–8515. doi: 10.1073/pnas.83.22.8511. [DOI] [PMC free article] [PubMed] [Google Scholar]
  28. Nakatani Y., Horikoshi M., Brenner M., Yamamoto T., Besnard F., Roeder R. G., Freese E. A downstream initiation element required for efficient TATA box binding and in vitro function of TFIID. Nature. 1990 Nov 1;348(6296):86–88. doi: 10.1038/348086a0. [DOI] [PubMed] [Google Scholar]
  29. Parslow T. G., Blair D. L., Murphy W. J., Granner D. K. Structure of the 5' ends of immunoglobulin genes: a novel conserved sequence. Proc Natl Acad Sci U S A. 1984 May;81(9):2650–2654. doi: 10.1073/pnas.81.9.2650. [DOI] [PMC free article] [PubMed] [Google Scholar]
  30. Parvin J. D., Sharp P. A. Identification of novel factors which bind specifically to the core promoter of the immunoglobulin heavy chain gene. J Biol Chem. 1991 Dec 5;266(34):22878–22886. [PubMed] [Google Scholar]
  31. Parvin J. D., Timmers H. T., Sharp P. A. Promoter specificity of basal transcription factors. Cell. 1992 Mar 20;68(6):1135–1144. doi: 10.1016/0092-8674(92)90084-p. [DOI] [PubMed] [Google Scholar]
  32. Purnell B. A., Emanuel P. A., Gilmour D. S. TFIID sequence recognition of the initiator and sequences farther downstream in Drosophila class II genes. Genes Dev. 1994 Apr 1;8(7):830–842. doi: 10.1101/gad.8.7.830. [DOI] [PubMed] [Google Scholar]
  33. Sawadogo M., Roeder R. G. Factors involved in specific transcription by human RNA polymerase II: analysis by a rapid and quantitative in vitro assay. Proc Natl Acad Sci U S A. 1985 Jul;82(13):4394–4398. doi: 10.1073/pnas.82.13.4394. [DOI] [PMC free article] [PubMed] [Google Scholar]
  34. Scheidereit C., Cromlish J. A., Gerster T., Kawakami K., Balmaceda C. G., Currie R. A., Roeder R. G. A human lymphoid-specific transcription factor that activates immunoglobulin genes is a homoeobox protein. Nature. 1988 Dec 8;336(6199):551–557. doi: 10.1038/336551a0. [DOI] [PubMed] [Google Scholar]
  35. Scheidereit C., Heguy A., Roeder R. G. Identification and purification of a human lymphoid-specific octamer-binding protein (OTF-2) that activates transcription of an immunoglobulin promoter in vitro. Cell. 1987 Dec 4;51(5):783–793. doi: 10.1016/0092-8674(87)90101-2. [DOI] [PubMed] [Google Scholar]
  36. Schubart D. B., Rolink A., Kosco-Vilbois M. H., Botteri F., Matthias P. B-cell-specific coactivator OBF-1/OCA-B/Bob1 required for immune response and germinal centre formation. Nature. 1996 Oct 10;383(6600):538–542. doi: 10.1038/383538a0. [DOI] [PubMed] [Google Scholar]
  37. Schäble K. F., Zachau H. G. The variable genes of the human immunoglobulin kappa locus. Biol Chem Hoppe Seyler. 1993 Nov;374(11):1001–1022. [PubMed] [Google Scholar]
  38. Sigvardsson M., Akerblad P., Leanderson T. Early B cell factor interacts with a subset of kappa promoters. J Immunol. 1996 May 15;156(10):3788–3796. [PubMed] [Google Scholar]
  39. Smale S. T., Baltimore D. The "initiator" as a transcription control element. Cell. 1989 Apr 7;57(1):103–113. doi: 10.1016/0092-8674(89)90176-1. [DOI] [PubMed] [Google Scholar]
  40. Staudt L. M., Lenardo M. J. Immunoglobulin gene transcription. Annu Rev Immunol. 1991;9:373–398. doi: 10.1146/annurev.iy.09.040191.002105. [DOI] [PubMed] [Google Scholar]
  41. Strubin M., Newell J. W., Matthias P. OBF-1, a novel B cell-specific coactivator that stimulates immunoglobulin promoter activity through association with octamer-binding proteins. Cell. 1995 Feb 10;80(3):497–506. doi: 10.1016/0092-8674(95)90500-6. [DOI] [PubMed] [Google Scholar]
  42. Sturm R. A., Das G., Herr W. The ubiquitous octamer-binding protein Oct-1 contains a POU domain with a homeo box subdomain. Genes Dev. 1988 Dec;2(12A):1582–1599. doi: 10.1101/gad.2.12a.1582. [DOI] [PubMed] [Google Scholar]
  43. Verrijzer C. P., Chen J. L., Yokomori K., Tjian R. Binding of TAFs to core elements directs promoter selectivity by RNA polymerase II. Cell. 1995 Jun 30;81(7):1115–1125. doi: 10.1016/s0092-8674(05)80016-9. [DOI] [PubMed] [Google Scholar]
  44. Verrijzer C. P., Yokomori K., Chen J. L., Tjian R. Drosophila TAFII150: similarity to yeast gene TSM-1 and specific binding to core promoter DNA. Science. 1994 May 13;264(5161):933–941. doi: 10.1126/science.8178153. [DOI] [PubMed] [Google Scholar]
  45. Weis L., Reinberg D. Transcription by RNA polymerase II: initiator-directed formation of transcription-competent complexes. FASEB J. 1992 Nov;6(14):3300–3309. doi: 10.1096/fasebj.6.14.1426767. [DOI] [PubMed] [Google Scholar]
  46. Wiley S. R., Kraus R. J., Zuo F., Murray E. E., Loritz K., Mertz J. E. SV40 early-to-late switch involves titration of cellular transcriptional repressors. Genes Dev. 1993 Nov;7(11):2206–2219. doi: 10.1101/gad.7.11.2206. [DOI] [PubMed] [Google Scholar]
  47. Yoza B. K., Roeder R. G. Identification of a novel factor that interacts with an immunoglobulin heavy-chain promoter and stimulates transcription in conjunction with the lymphoid cell-specific factor OTF2. Mol Cell Biol. 1990 May;10(5):2145–2153. doi: 10.1128/mcb.10.5.2145. [DOI] [PMC free article] [PubMed] [Google Scholar]
  48. Zwilling S., Annweiler A., Wirth T. The POU domains of the Oct1 and Oct2 transcription factors mediate specific interaction with TBP. Nucleic Acids Res. 1994 May 11;22(9):1655–1662. doi: 10.1093/nar/22.9.1655. [DOI] [PMC free article] [PubMed] [Google Scholar]

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