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. 1998 Sep 15;26(18):4100–4107. doi: 10.1093/nar/26.18.4100

The N-terminal domain unique to the long form of the Brn-3a transcription factor is essential to protect neuronal cells from apoptosis and for the activation of Bbcl-2 gene expression.

M D Smith 1, S J Dawson 1, L M Boxer 1, D S Latchman 1
PMCID: PMC147830  PMID: 9722627

Abstract

The ability of the POU family transcription factor Brn-3a to stimulate neurite outgrowth and the expression of the genes encoding neuronal proteins such as the neurofilaments and SNAP-25 has previously been shown to be dependent upon the C-terminal POU domain which can mediate both DNA binding and transcriptional activation. We show here, however, that the ability of Brn-3a to activate Bcl-2 expression and protect neuronal cells from apoptosis (programmed cell death) requires a distinct N-terminal activation domain. Bcl-2 gene activation and protection from apoptosis are thus produced only by the long form of Brn-3a which contains this domain and not by a naturally occurring short form lacking this domain or by the isolated POU domain, although all these forms of Brn-3a can stimulate neurite outgrowth. Hence Brn-3a is a multi-functional transcription factor with different regions of the factor mediating its different effects and two distinct forms with different properties being generated by alternative splicing.

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Selected References

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  1. Budhram-Mahadeo V., Morris P. J., Lakin N. D., Theil T., Ching G. Y., Lillycrop K. A., Möröy T., Liem R. K., Latchman D. S. Activation of the alpha-internexin promoter by the Brn-3a transcription factor is dependent on the N-terminal region of the protein. J Biol Chem. 1995 Feb 10;270(6):2853–2858. doi: 10.1074/jbc.270.6.2853. [DOI] [PubMed] [Google Scholar]
  2. Budhram-Mahadeo V., Theil T., Morris P. J., Lillycrop K. A., Moroy T., Latchman D. S. The DNA target site for the Brn-3 POU family transcription factors can confer responsiveness to cyclic AMP and removal of serum in neuronal cells. Nucleic Acids Res. 1994 Aug 11;22(15):3092–3098. doi: 10.1093/nar/22.15.3092. [DOI] [PMC free article] [PubMed] [Google Scholar]
  3. Dawson S. J., Morris P. J., Latchman D. S. A single amino acid change converts an inhibitory transcription factor into an activator. J Biol Chem. 1996 May 17;271(20):11631–11633. doi: 10.1074/jbc.271.20.11631. [DOI] [PubMed] [Google Scholar]
  4. Erkman L., McEvilly R. J., Luo L., Ryan A. K., Hooshmand F., O'Connell S. M., Keithley E. M., Rapaport D. H., Ryan A. F., Rosenfeld M. G. Role of transcription factors Brn-3.1 and Brn-3.2 in auditory and visual system development. Nature. 1996 Jun 13;381(6583):603–606. doi: 10.1038/381603a0. [DOI] [PubMed] [Google Scholar]
  5. Fedtsova N. G., Turner E. E. Brn-3.0 expression identifies early post-mitotic CNS neurons and sensory neural precursors. Mech Dev. 1995 Nov;53(3):291–304. doi: 10.1016/0925-4773(95)00435-1. [DOI] [PubMed] [Google Scholar]
  6. Gavrieli Y., Sherman Y., Ben-Sasson S. A. Identification of programmed cell death in situ via specific labeling of nuclear DNA fragmentation. J Cell Biol. 1992 Nov;119(3):493–501. doi: 10.1083/jcb.119.3.493. [DOI] [PMC free article] [PubMed] [Google Scholar]
  7. Gerrero M. R., McEvilly R. J., Turner E., Lin C. R., O'Connell S., Jenne K. J., Hobbs M. V., Rosenfeld M. G. Brn-3.0: a POU-domain protein expressed in the sensory, immune, and endocrine systems that functions on elements distinct from known octamer motifs. Proc Natl Acad Sci U S A. 1993 Nov 15;90(22):10841–10845. doi: 10.1073/pnas.90.22.10841. [DOI] [PMC free article] [PubMed] [Google Scholar]
  8. 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]
  9. Howard M. K., Burke L. C., Mailhos C., Pizzey A., Gilbert C. S., Lawson W. D., Collins M. K., Thomas N. S., Latchman D. S. Cell cycle arrest of proliferating neuronal cells by serum deprivation can result in either apoptosis or differentiation. J Neurochem. 1993 May;60(5):1783–1791. doi: 10.1111/j.1471-4159.1993.tb13404.x. [DOI] [PubMed] [Google Scholar]
  10. Lakin N. D., Morris P. J., Theil T., Sato T. N., Möröy T., Wilson M. C., Latchman D. S. Regulation of neurite outgrowth and SNAP-25 gene expression by the Brn-3a transcription factor. J Biol Chem. 1995 Jun 30;270(26):15858–15863. doi: 10.1074/jbc.270.26.15858. [DOI] [PubMed] [Google Scholar]
  11. Lillycrop K. A., Budrahan V. S., Lakin N. D., Terrenghi G., Wood J. N., Polak J. M., Latchman D. S. A novel POU family transcription factor is closely related to Brn-3 but has a distinct expression pattern in neuronal cells. Nucleic Acids Res. 1992 Oct 11;20(19):5093–5096. doi: 10.1093/nar/20.19.5093. [DOI] [PMC free article] [PubMed] [Google Scholar]
  12. Liu Y. Z., Dawson S. J., Latchman D. S. Alternative splicing of the Brn-3a and Brn-3b transcription factor RNAs is regulated in neuronal cells. J Mol Neurosci. 1996 Spring;7(1):77–85. doi: 10.1007/BF02736850. [DOI] [PubMed] [Google Scholar]
  13. Luckow B., Schütz G. CAT constructions with multiple unique restriction sites for the functional analysis of eukaryotic promoters and regulatory elements. Nucleic Acids Res. 1987 Jul 10;15(13):5490–5490. doi: 10.1093/nar/15.13.5490. [DOI] [PMC free article] [PubMed] [Google Scholar]
  14. 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]
  15. McEvilly R. J., Erkman L., Luo L., Sawchenko P. E., Ryan A. F., Rosenfeld M. G. Requirement for Brn-3.0 in differentiation and survival of sensory and motor neurons. Nature. 1996 Dec 12;384(6609):574–577. doi: 10.1038/384574a0. [DOI] [PubMed] [Google Scholar]
  16. Morgenstern J. P., Land H. Advanced mammalian gene transfer: high titre retroviral vectors with multiple drug selection markers and a complementary helper-free packaging cell line. Nucleic Acids Res. 1990 Jun 25;18(12):3587–3596. doi: 10.1093/nar/18.12.3587. [DOI] [PMC free article] [PubMed] [Google Scholar]
  17. Morris P. J., Dawson S. J., Wilson M. C., Latchman D. S. A single residue within the homeodomain of the Brn-3 POU family transcription factors determines whether they activate or repress the SNAP-25 promoter. Neuroreport. 1997 May 27;8(8):2041–2045. doi: 10.1097/00001756-199705260-00047. [DOI] [PubMed] [Google Scholar]
  18. Morris P. J., Lakin N. D., Dawson S. J., Ryabinin A. E., Kilimann M. W., Wilson M. C., Latchman D. S. Differential regulation of genes encoding synaptic proteins by members of the Brn-3 subfamily of POU transcription factors. Brain Res Mol Brain Res. 1996 Dec 31;43(1-2):279–285. doi: 10.1016/s0169-328x(96)00207-0. [DOI] [PubMed] [Google Scholar]
  19. Morris P. J., Theil T., Ring C. J., Lillycrop K. A., Moroy T., Latchman D. S. The opposite and antagonistic effects of the closely related POU family transcription factors Brn-3a and Brn-3b on the activity of a target promoter are dependent on differences in the POU domain. Mol Cell Biol. 1994 Oct;14(10):6907–6914. doi: 10.1128/mcb.14.10.6907. [DOI] [PMC free article] [PubMed] [Google Scholar]
  20. Ninkina N. N., Stevens G. E., Wood J. N., Richardson W. D. A novel Brn3-like POU transcription factor expressed in subsets of rat sensory and spinal cord neurons. Nucleic Acids Res. 1993 Jul 11;21(14):3175–3182. doi: 10.1093/nar/21.14.3175. [DOI] [PMC free article] [PubMed] [Google Scholar]
  21. Osen-Sand A., Catsicas M., Staple J. K., Jones K. A., Ayala G., Knowles J., Grenningloh G., Catsicas S. Inhibition of axonal growth by SNAP-25 antisense oligonucleotides in vitro and in vivo. Nature. 1993 Jul 29;364(6436):445–448. doi: 10.1038/364445a0. [DOI] [PubMed] [Google Scholar]
  22. Ryan A. K., Rosenfeld M. G. POU domain family values: flexibility, partnerships, and developmental codes. Genes Dev. 1997 May 15;11(10):1207–1225. doi: 10.1101/gad.11.10.1207. [DOI] [PubMed] [Google Scholar]
  23. Sellins K. S., Cohen J. J. Gene induction by gamma-irradiation leads to DNA fragmentation in lymphocytes. J Immunol. 1987 Nov 15;139(10):3199–3206. [PubMed] [Google Scholar]
  24. Smith M. D., Dawson S. J., Latchman D. S. Inhibition of neuronal process outgrowth and neuronal specific gene activation by the Brn-3b transcription factor. J Biol Chem. 1997 Jan 10;272(2):1382–1388. doi: 10.1074/jbc.272.2.1382. [DOI] [PubMed] [Google Scholar]
  25. Smith M. D., Dawson S. J., Latchman D. S. The Brn-3a transcription factor induces neuronal process outgrowth and the coordinate expression of genes encoding synaptic proteins. Mol Cell Biol. 1997 Jan;17(1):345–354. doi: 10.1128/mcb.17.1.345. [DOI] [PMC free article] [PubMed] [Google Scholar]
  26. Smith M. D., Ensor E. A., Coffin R. S., Boxer L. M., Latchman D. S. Bcl-2 transcription from the proximal P2 promoter is activated in neuronal cells by the Brn-3a POU family transcription factor. J Biol Chem. 1998 Jul 3;273(27):16715–16722. doi: 10.1074/jbc.273.27.16715. [DOI] [PubMed] [Google Scholar]
  27. Smith M. D., Morris P. J., Dawson S. J., Schwartz M. L., Schlaepfer W. W., Latchman D. S. Coordinate induction of the three neurofilament genes by the Brn-3a transcription factor. J Biol Chem. 1997 Aug 22;272(34):21325–21333. doi: 10.1074/jbc.272.34.21325. [DOI] [PubMed] [Google Scholar]
  28. Theil T., McLean-Hunter S., Zörnig M., Möröy T. Mouse Brn-3 family of POU transcription factors: a new aminoterminal domain is crucial for the oncogenic activity of Brn-3a. Nucleic Acids Res. 1993 Dec 25;21(25):5921–5929. doi: 10.1093/nar/21.25.5921. [DOI] [PMC free article] [PubMed] [Google Scholar]
  29. Theil T., Zechner U., Klett C., Adolph S., Möröy T. Chromosomal localization and sequences of the murine Brn-3 family of developmental control genes. Cytogenet Cell Genet. 1994;66(4):267–271. doi: 10.1159/000133709. [DOI] [PubMed] [Google Scholar]
  30. Turner E. E., Jenne K. J., Rosenfeld M. G. Brn-3.2: a Brn-3-related transcription factor with distinctive central nervous system expression and regulation by retinoic acid. Neuron. 1994 Jan;12(1):205–218. doi: 10.1016/0896-6273(94)90164-3. [DOI] [PubMed] [Google Scholar]
  31. 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]
  32. Wilson B. E., Mochon E., Boxer L. M. Induction of bcl-2 expression by phosphorylated CREB proteins during B-cell activation and rescue from apoptosis. Mol Cell Biol. 1996 Oct;16(10):5546–5556. doi: 10.1128/mcb.16.10.5546. [DOI] [PMC free article] [PubMed] [Google Scholar]
  33. 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]
  34. Xiang M., Gan L., Zhou L., Klein W. H., Nathans J. Targeted deletion of the mouse POU domain gene Brn-3a causes selective loss of neurons in the brainstem and trigeminal ganglion, uncoordinated limb movement, and impaired suckling. Proc Natl Acad Sci U S A. 1996 Oct 15;93(21):11950–11955. doi: 10.1073/pnas.93.21.11950. [DOI] [PMC free article] [PubMed] [Google Scholar]

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