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. 1998 May 15;26(10):2464–2472. doi: 10.1093/nar/26.10.2464

Adjacent proline residues in the inhibitory domain of the Oct-2 transcription factor play distinct functional roles.

Y Z Liu 1, I K Lee 1, I Locke 1, S J Dawson 1, D S Latchman 1
PMCID: PMC147571  PMID: 9580701

Abstract

A 40 amino acid region of Oct-2 from amino acids 142 to 181 functions as an active repressor domain capable of inhibiting both basal activity and activation of promoters containing a TATA box, but not of those that contain an initiator element. Based on our observation that the equivalent region of the closely related Oct-1 factor does not act as an inhibitory domain, we have mutated specific residues in the Oct-2 domain in an attempt to probe their importance in repressor domain function. Although mutations of several residues have no or minimal effect, mutation of proline 175 to arginine abolishes the ability to inhibit both basal and activated transcription. In contrast, mutation of proline 174 to arginine confers upon the domain the ability to repress activation of an initiator-containing promoter by acidic activation domains, and also suppresses the effect of the proline 175 mutation. Hence, adjacent proline residues play key roles in the functioning of the inhibitory domain and in limiting its specificity to TATA-box-containing promoters.

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Selected References

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

  1. Abken H., Reifenrath B. A procedure to standardize CAT reporter gene assay. Nucleic Acids Res. 1992 Jul 11;20(13):3527–3527. doi: 10.1093/nar/20.13.3527. [DOI] [PMC free article] [PubMed] [Google Scholar]
  2. Annweiler A., Zwilling S., Wirth T. Functional differences between the Oct2 transactivation domains determine the transactivation potential of individual Oct2 isoforms. Nucleic Acids Res. 1994 Oct 11;22(20):4250–4258. doi: 10.1093/nar/22.20.4250. [DOI] [PMC free article] [PubMed] [Google Scholar]
  3. Austin R. J., Biggin M. D. A domain of the even-skipped protein represses transcription by preventing TFIID binding to a promoter: repression by cooperative blocking. Mol Cell Biol. 1995 Sep;15(9):4683–4693. doi: 10.1128/mcb.15.9.4683. [DOI] [PMC free article] [PubMed] [Google Scholar]
  4. Bradford M. M. A rapid and sensitive method for the quantitation of microgram quantities of protein utilizing the principle of protein-dye binding. Anal Biochem. 1976 May 7;72:248–254. doi: 10.1016/0003-2697(76)90527-3. [DOI] [PubMed] [Google Scholar]
  5. 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]
  6. Cowell I. G., Hurst H. C. Transcriptional repression by the human bZIP factor E4BP4: definition of a minimal repression domain. Nucleic Acids Res. 1994 Jan 11;22(1):59–65. doi: 10.1093/nar/22.1.59. [DOI] [PMC free article] [PubMed] [Google Scholar]
  7. Cowell I. G. Repression versus activation in the control of gene transcription. Trends Biochem Sci. 1994 Jan;19(1):38–42. doi: 10.1016/0968-0004(94)90172-4. [DOI] [PubMed] [Google Scholar]
  8. Dawson S. J., Yoon S. O., Chikaraishi D. M., Lillycrop K. A., Latchman D. S. The Oct-2 transcription factor represses tyrosine hydroxylase expression via a heptamer TAATGARAT-like motif in the gene promoter. Nucleic Acids Res. 1994 Mar 25;22(6):1023–1028. doi: 10.1093/nar/22.6.1023. [DOI] [PMC free article] [PubMed] [Google Scholar]
  9. Deans Z. C., Dawson S. J., Kilimann M. W., Wallace D., Wilson M. C., Latchman D. S. Differential regulation of genes encoding synaptic proteins by the Oct-2 transcription factor. Brain Res Mol Brain Res. 1997 Nov;51(1-2):1–7. doi: 10.1016/s0169-328x(97)00204-0. [DOI] [PubMed] [Google Scholar]
  10. Dent C. L., Lillycrop K. A., Estridge J. K., Thomas N. S., Latchman D. S. The B-cell and neuronal forms of the octamer-binding protein Oct-2 differ in DNA-binding specificity and functional activity. Mol Cell Biol. 1991 Aug;11(8):3925–3930. doi: 10.1128/mcb.11.8.3925. [DOI] [PMC free article] [PubMed] [Google Scholar]
  11. Friedl E. M., Matthias P. Transcriptional activation and repression, two properties of the lymphoid-specific transcription factor Oct-2a. Eur J Biochem. 1995 Nov 15;234(1):308–316. doi: 10.1111/j.1432-1033.1995.308_c.x. [DOI] [PubMed] [Google Scholar]
  12. Gay R. D., Dawson S. J., Latchman D. S. The different inhibitory domains of the Oct-2 transcription factor have distinct functional activities. FEBS Lett. 1997 Oct 20;416(2):135–138. doi: 10.1016/s0014-5793(97)01184-8. [DOI] [PubMed] [Google Scholar]
  13. Gerster T., Balmaceda C. G., Roeder R. G. The cell type-specific octamer transcription factor OTF-2 has two domains required for the activation of transcription. EMBO J. 1990 May;9(5):1635–1643. doi: 10.1002/j.1460-2075.1990.tb08283.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  14. Gray S., Szymanski P., Levine M. Short-range repression permits multiple enhancers to function autonomously within a complex promoter. Genes Dev. 1994 Aug 1;8(15):1829–1838. doi: 10.1101/gad.8.15.1829. [DOI] [PubMed] [Google Scholar]
  15. Han K., Manley J. L. Transcriptional repression by the Drosophila even-skipped protein: definition of a minimal repression domain. Genes Dev. 1993 Mar;7(3):491–503. doi: 10.1101/gad.7.3.491. [DOI] [PubMed] [Google Scholar]
  16. Hanna-Rose W., Hansen U. Active repression mechanisms of eukaryotic transcription repressors. Trends Genet. 1996 Jun;12(6):229–234. doi: 10.1016/0168-9525(96)10022-6. [DOI] [PubMed] [Google Scholar]
  17. Hatzopoulos A. K., Stoykova A. S., Erselius J. R., Goulding M., Neuman T., Gruss P. Structure and expression of the mouse Oct2a and Oct2b, two differentially spliced products of the same gene. Development. 1990 Jun;109(2):349–362. doi: 10.1242/dev.109.2.349. [DOI] [PubMed] [Google Scholar]
  18. He X., Treacy M. N., Simmons D. M., Ingraham H. A., Swanson L. W., Rosenfeld M. G. Expression of a large family of POU-domain regulatory genes in mammalian brain development. Nature. 1989 Jul 6;340(6228):35–41. doi: 10.1038/340035a0. [DOI] [PubMed] [Google Scholar]
  19. Johnston S. A., Salmeron J. M., Jr, Dincher S. S. Interaction of positive and negative regulatory proteins in the galactose regulon of yeast. Cell. 1987 Jul 3;50(1):143–146. doi: 10.1016/0092-8674(87)90671-4. [DOI] [PubMed] [Google Scholar]
  20. Johnston S. A., Salmeron J. M., Jr, Dincher S. S. Interaction of positive and negative regulatory proteins in the galactose regulon of yeast. Cell. 1987 Jul 3;50(1):143–146. doi: 10.1016/0092-8674(87)90671-4. [DOI] [PubMed] [Google Scholar]
  21. Kemp L. M., Dent C. L., Latchman D. S. Octamer motif mediates transcriptional repression of HSV immediate-early genes and octamer-containing cellular promoters in neuronal cells. Neuron. 1990 Feb;4(2):215–222. doi: 10.1016/0896-6273(90)90096-x. [DOI] [PubMed] [Google Scholar]
  22. König H., Pfisterer P., Corcoran L. M., Wirth T. Identification of CD36 as the first gene dependent on the B-cell differentiation factor Oct-2. Genes Dev. 1995 Jul 1;9(13):1598–1607. doi: 10.1101/gad.9.13.1598. [DOI] [PubMed] [Google Scholar]
  23. Licht J. D., Hanna-Rose W., Reddy J. C., English M. A., Ro M., Grossel M., Shaknovich R., Hansen U. Mapping and mutagenesis of the amino-terminal transcriptional repression domain of the Drosophila Krüppel protein. Mol Cell Biol. 1994 Jun;14(6):4057–4066. doi: 10.1128/mcb.14.6.4057. [DOI] [PMC free article] [PubMed] [Google Scholar]
  24. Lillycrop K. A., Dawson S. J., Estridge J. K., Gerster T., Matthias P., Latchman D. S. Repression of a herpes simplex virus immediate-early promoter by the Oct-2 transcription factor is dependent on an inhibitory region at the N terminus of the protein. Mol Cell Biol. 1994 Nov;14(11):7633–7642. doi: 10.1128/mcb.14.11.7633. [DOI] [PMC free article] [PubMed] [Google Scholar]
  25. Lillycrop K. A., Dent C. L., Wheatley S. C., Beech M. N., Ninkina N. N., Wood J. N., Latchman D. S. The octamer-binding protein Oct-2 represses HSV immediate-early genes in cell lines derived from latently infectable sensory neurons. Neuron. 1991 Sep;7(3):381–390. doi: 10.1016/0896-6273(91)90290-g. [DOI] [PubMed] [Google Scholar]
  26. Lillycrop K. A., Latchman D. S. Alternative splicing of the Oct-2 transcription factor RNA is differentially regulated in neuronal cells and B cells and results in protein isoforms with opposite effects on the activity of octamer/TAATGARAT-containing promoters. J Biol Chem. 1992 Dec 15;267(35):24960–24965. [PubMed] [Google Scholar]
  27. Lillycrop K. A., Latchman D. S. The inhibitory domain in the Oct-2 transcription factor represses gene activity in a cell type-specific and promoter-independent manner. Mol Biol Rep. 1995;21(2):87–94. doi: 10.1007/BF00986498. [DOI] [PubMed] [Google Scholar]
  28. Liu Y. Z., Dawson S. J., Gerster T., Friedl E., Pengue G., Matthias P., Lania L., Latchman D. S. The ability of the inhibitory domain of the POU family transcription factor Oct-2 to interfere with promoter activation by different classes of activation domains is dependent upon the nature of the basal promoter elements. J Biol Chem. 1996 Aug 23;271(34):20853–20860. doi: 10.1074/jbc.271.34.20853. [DOI] [PubMed] [Google Scholar]
  29. MACPHERSON I., STOKER M. Polyoma transformation of hamster cell clones--an investigation of genetic factors affecting cell competence. Virology. 1962 Feb;16:147–151. doi: 10.1016/0042-6822(62)90290-8. [DOI] [PubMed] [Google Scholar]
  30. Madden S. L., Cook D. M., Rauscher F. J., 3rd A structure-function analysis of transcriptional repression mediated by the WT1, Wilms' tumor suppressor protein. Oncogene. 1993 Jul;8(7):1713–1720. [PubMed] [Google Scholar]
  31. Müller-Immerglück M. M., Schaffner W., Matthias P. Transcription factor Oct-2A contains functionally redundant activating domains and works selectively from a promoter but not from a remote enhancer position in non-lymphoid (HeLa) cells. EMBO J. 1990 May;9(5):1625–1634. doi: 10.1002/j.1460-2075.1990.tb08282.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  32. Pengue G., Lania L. Krüppel-associated box-mediated repression of RNA polymerase II promoters is influenced by the arrangement of basal promoter elements. Proc Natl Acad Sci U S A. 1996 Feb 6;93(3):1015–1020. doi: 10.1073/pnas.93.3.1015. [DOI] [PMC free article] [PubMed] [Google Scholar]
  33. Pfisterer P., König H., Hess J., Lipowsky G., Haendler B., Schleuning W. D., Wirth T. CRISP-3, a protein with homology to plant defense proteins, is expressed in mouse B cells under the control of Oct2. Mol Cell Biol. 1996 Nov;16(11):6160–6168. doi: 10.1128/mcb.16.11.6160. [DOI] [PMC free article] [PubMed] [Google Scholar]
  34. Pugh B. F., Tjian R. Mechanism of transcriptional activation by Sp1: evidence for coactivators. Cell. 1990 Jun 29;61(7):1187–1197. doi: 10.1016/0092-8674(90)90683-6. [DOI] [PubMed] [Google Scholar]
  35. Sadowski I., Ptashne M. A vector for expressing GAL4(1-147) fusions in mammalian cells. Nucleic Acids Res. 1989 Sep 25;17(18):7539–7539. doi: 10.1093/nar/17.18.7539. [DOI] [PMC free article] [PubMed] [Google Scholar]
  36. Saha S., Brickman J. M., Lehming N., Ptashne M. New eukaryotic transcriptional repressors. Nature. 1993 Jun 17;363(6430):648–652. doi: 10.1038/363648a0. [DOI] [PubMed] [Google Scholar]
  37. Schöler H. R., Hatzopoulos A. K., Balling R., Suzuki N., Gruss P. A family of octamer-specific proteins present during mouse embryogenesis: evidence for germline-specific expression of an Oct factor. EMBO J. 1989 Sep;8(9):2543–2550. doi: 10.1002/j.1460-2075.1989.tb08392.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  38. Sen R., Baltimore D. Multiple nuclear factors interact with the immunoglobulin enhancer sequences. Cell. 1986 Aug 29;46(5):705–716. doi: 10.1016/0092-8674(86)90346-6. [DOI] [PubMed] [Google Scholar]
  39. Shi Y., Seto E., Chang L. S., Shenk T. Transcriptional repression by YY1, a human GLI-Krüppel-related protein, and relief of repression by adenovirus E1A protein. Cell. 1991 Oct 18;67(2):377–388. doi: 10.1016/0092-8674(91)90189-6. [DOI] [PubMed] [Google Scholar]
  40. 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]
  41. Staudt L. M., Clerc R. G., Singh H., LeBowitz J. H., Sharp P. A., Baltimore D. Cloning of a lymphoid-specific cDNA encoding a protein binding the regulatory octamer DNA motif. Science. 1988 Jul 29;241(4865):577–580. doi: 10.1126/science.3399892. [DOI] [PubMed] [Google Scholar]
  42. Stoykova A. S., Sterrer S., Erselius J. R., Hatzopoulos A. K., Gruss P. Mini-Oct and Oct-2c: two novel, functionally diverse murine Oct-2 gene products are differentially expressed in the CNS. Neuron. 1992 Mar;8(3):541–558. doi: 10.1016/0896-6273(92)90282-i. [DOI] [PubMed] [Google Scholar]
  43. Verrijzer C. P., Van der Vliet P. C. POU domain transcription factors. Biochim Biophys Acta. 1993 Apr 29;1173(1):1–21. doi: 10.1016/0167-4781(93)90237-8. [DOI] [PubMed] [Google Scholar]
  44. Wegner M., Drolet D. W., Rosenfeld M. G. POU-domain proteins: structure and function of developmental regulators. Curr Opin Cell Biol. 1993 Jun;5(3):488–498. doi: 10.1016/0955-0674(93)90015-i. [DOI] [PubMed] [Google Scholar]
  45. Wirth T., Priess A., Annweiler A., Zwilling S., Oeler B. Multiple Oct2 isoforms are generated by alternative splicing. Nucleic Acids Res. 1991 Jan 11;19(1):43–51. doi: 10.1093/nar/19.1.43. [DOI] [PMC free article] [PubMed] [Google Scholar]
  46. Wood J. N., Bevan S. J., Coote P. R., Dunn P. M., Harmar A., Hogan P., Latchman D. S., Morrison C., Rougon G., Theveniau M. Novel cell lines display properties of nociceptive sensory neurons. Proc Biol Sci. 1990 Sep 22;241(1302):187–194. doi: 10.1098/rspb.1990.0084. [DOI] [PubMed] [Google Scholar]
  47. Yamamoto H., Lamphier M. S., Fujita T., Taniguchi T., Harada H. The oncogenic transcription factor IRF-2 possesses a transcriptional repression and a latent activation domain. Oncogene. 1994 May;9(5):1423–1428. [PubMed] [Google Scholar]
  48. Zeleznik-Le N. J., Harden A. M., Rowley J. D. 11q23 translocations split the "AT-hook" cruciform DNA-binding region and the transcriptional repression domain from the activation domain of the mixed-lineage leukemia (MLL) gene. Proc Natl Acad Sci U S A. 1994 Oct 25;91(22):10610–10614. doi: 10.1073/pnas.91.22.10610. [DOI] [PMC free article] [PubMed] [Google Scholar]

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