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. 1997 Jan 15;16(2):384–395. doi: 10.1093/emboj/16.2.384

Conversion of ectoderm into a neural fate by ATH-3, a vertebrate basic helix-loop-helix gene homologous to Drosophila proneural gene atonal.

K Takebayashi 1, S Takahashi 1, C Yokota 1, H Tsuda 1, S Nakanishi 1, M Asashima 1, R Kageyama 1
PMCID: PMC1169643  PMID: 9029157

Abstract

We have isolated a novel basic helix-loop-helix (bHLH) gene homologous to the Drosophila proneural gene atonal, termed ATH-3, from Xenopus and mouse. ATH-3 is expressed in the developing nervous system, with high levels of expression in the brain, retina and cranial ganglions. Injection of ATH-3 RNA into Xenopus embryos dramatically expands the neural tube and induces ectopic neural tissues in the epidermis but inhibits non-neural development. This ATH-3-induced neural hyperplasia does not require cell division, indicating that surrounding cells which are normally non-neural types adopt a neural fate. In a Xenopus animal cap assay, ATH-3 is able to convert ectodermal cells into neurons expressing anterior markers without inducing mesoderm. Interestingly, a single amino acid change from Ser to Asp in the basic region, which mimics phosphorylation of Ser, severely impairs the anterior marker-inducing ability without affecting general neurogenic activities. These results provide evidence that ATH-3 can directly convert non-neural or undetermined cells into a neural fate, and suggest that the Ser residue in the basic region may be critical for the regulation of ATH-3 activity by phosphorylation.

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Selected References

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

  1. Akazawa C., Sasai Y., Nakanishi S., Kageyama R. Molecular characterization of a rat negative regulator with a basic helix-loop-helix structure predominantly expressed in the developing nervous system. J Biol Chem. 1992 Oct 25;267(30):21879–21885. [PubMed] [Google Scholar]
  2. Bader D., Masaki T., Fischman D. A. Immunochemical analysis of myosin heavy chain during avian myogenesis in vivo and in vitro. J Cell Biol. 1982 Dec;95(3):763–770. doi: 10.1083/jcb.95.3.763. [DOI] [PMC free article] [PubMed] [Google Scholar]
  3. Benezra R., Davis R. L., Lockshon D., Turner D. L., Weintraub H. The protein Id: a negative regulator of helix-loop-helix DNA binding proteins. Cell. 1990 Apr 6;61(1):49–59. doi: 10.1016/0092-8674(90)90214-y. [DOI] [PubMed] [Google Scholar]
  4. Ferreiro B., Kintner C., Zimmerman K., Anderson D., Harris W. A. XASH genes promote neurogenesis in Xenopus embryos. Development. 1994 Dec;120(12):3649–3655. doi: 10.1242/dev.120.12.3649. [DOI] [PubMed] [Google Scholar]
  5. Guillemot F., Lo L. C., Johnson J. E., Auerbach A., Anderson D. J., Joyner A. L. Mammalian achaete-scute homolog 1 is required for the early development of olfactory and autonomic neurons. Cell. 1993 Nov 5;75(3):463–476. doi: 10.1016/0092-8674(93)90381-y. [DOI] [PubMed] [Google Scholar]
  6. Halder G., Callaerts P., Gehring W. J. Induction of ectopic eyes by targeted expression of the eyeless gene in Drosophila. Science. 1995 Mar 24;267(5205):1788–1792. doi: 10.1126/science.7892602. [DOI] [PubMed] [Google Scholar]
  7. Harland R. M. In situ hybridization: an improved whole-mount method for Xenopus embryos. Methods Cell Biol. 1991;36:685–695. doi: 10.1016/s0091-679x(08)60307-6. [DOI] [PubMed] [Google Scholar]
  8. Harris W. A., Hartenstein V. Neuronal determination without cell division in Xenopus embryos. Neuron. 1991 Apr;6(4):499–515. doi: 10.1016/0896-6273(91)90053-3. [DOI] [PubMed] [Google Scholar]
  9. Hatada S., Kinoshita M., Noda M., Asashima M. Identification of a Xenopus glutamine synthetase gene abundantly expressed in the embryonic nervous system but not in adult brain. FEBS Lett. 1995 Sep 11;371(3):287–292. doi: 10.1016/0014-5793(95)00913-t. [DOI] [PubMed] [Google Scholar]
  10. Hemmati-Brivanlou A., Melton D. A. Inhibition of activin receptor signaling promotes neuralization in Xenopus. Cell. 1994 Apr 22;77(2):273–281. doi: 10.1016/0092-8674(94)90319-0. [DOI] [PubMed] [Google Scholar]
  11. Hill R. E., Favor J., Hogan B. L., Ton C. C., Saunders G. F., Hanson I. M., Prosser J., Jordan T., Hastie N. D., van Heyningen V. Mouse small eye results from mutations in a paired-like homeobox-containing gene. Nature. 1991 Dec 19;354(6354):522–525. doi: 10.1038/354522a0. [DOI] [PubMed] [Google Scholar]
  12. Hopwood N. D., Pluck A., Gurdon J. B. A Xenopus mRNA related to Drosophila twist is expressed in response to induction in the mesoderm and the neural crest. Cell. 1989 Dec 1;59(5):893–903. doi: 10.1016/0092-8674(89)90612-0. [DOI] [PubMed] [Google Scholar]
  13. Isaka F., Shimizu C., Nakanishi S., Kageyama R. Genetic mapping of four mouse bHLH genes related to Drosophila proneural gene atonal. Genomics. 1996 Nov 1;37(3):400–402. doi: 10.1006/geno.1996.0577. [DOI] [PubMed] [Google Scholar]
  14. Ishibashi M., Ang S. L., Shiota K., Nakanishi S., Kageyama R., Guillemot F. Targeted disruption of mammalian hairy and Enhancer of split homolog-1 (HES-1) leads to up-regulation of neural helix-loop-helix factors, premature neurogenesis, and severe neural tube defects. Genes Dev. 1995 Dec 15;9(24):3136–3148. doi: 10.1101/gad.9.24.3136. [DOI] [PubMed] [Google Scholar]
  15. Ishibashi M., Moriyoshi K., Sasai Y., Shiota K., Nakanishi S., Kageyama R. Persistent expression of helix-loop-helix factor HES-1 prevents mammalian neural differentiation in the central nervous system. EMBO J. 1994 Apr 15;13(8):1799–1805. doi: 10.1002/j.1460-2075.1994.tb06448.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  16. Jan Y. N., Jan L. Y. HLH proteins, fly neurogenesis, and vertebrate myogenesis. Cell. 1993 Dec 3;75(5):827–830. doi: 10.1016/0092-8674(93)90525-u. [DOI] [PubMed] [Google Scholar]
  17. Jarman A. P., Grau Y., Jan L. Y., Jan Y. N. atonal is a proneural gene that directs chordotonal organ formation in the Drosophila peripheral nervous system. Cell. 1993 Jul 2;73(7):1307–1321. doi: 10.1016/0092-8674(93)90358-w. [DOI] [PubMed] [Google Scholar]
  18. Jarman A. P., Grell E. H., Ackerman L., Jan L. Y., Jan Y. N. Atonal is the proneural gene for Drosophila photoreceptors. Nature. 1994 Jun 2;369(6479):398–400. doi: 10.1038/369398a0. [DOI] [PubMed] [Google Scholar]
  19. Johnson J. E., Birren S. J., Anderson D. J. Two rat homologues of Drosophila achaete-scute specifically expressed in neuronal precursors. Nature. 1990 Aug 30;346(6287):858–861. doi: 10.1038/346858a0. [DOI] [PubMed] [Google Scholar]
  20. Kageyama R., Sasai Y., Akazawa C., Ishibashi M., Takebayashi K., Shimizu C., Tomita K., Nakanishi S. Regulation of mammalian neural development by helix-loop-helix transcription factors. Crit Rev Neurobiol. 1995;9(2-3):177–188. [PubMed] [Google Scholar]
  21. Kohl N. E., Legouy E., DePinho R. A., Nisen P. D., Smith R. K., Gee C. E., Alt F. W. Human N-myc is closely related in organization and nucleotide sequence to c-myc. Nature. 1986 Jan 2;319(6048):73–77. doi: 10.1038/319073a0. [DOI] [PubMed] [Google Scholar]
  22. Krieg P. A., Melton D. A. Functional messenger RNAs are produced by SP6 in vitro transcription of cloned cDNAs. Nucleic Acids Res. 1984 Sep 25;12(18):7057–7070. doi: 10.1093/nar/12.18.7057. [DOI] [PMC free article] [PubMed] [Google Scholar]
  23. Krieg P. A., Varnum S. M., Wormington W. M., Melton D. A. The mRNA encoding elongation factor 1-alpha (EF-1 alpha) is a major transcript at the midblastula transition in Xenopus. Dev Biol. 1989 May;133(1):93–100. doi: 10.1016/0012-1606(89)90300-x. [DOI] [PubMed] [Google Scholar]
  24. Kume H., Maruyama K., Tomita T., Iwatsubo T., Saido T. C., Obata K. Molecular cloning of a novel basic helix-loop-helix protein from the rat brain. Biochem Biophys Res Commun. 1996 Feb 15;219(2):526–530. doi: 10.1006/bbrc.1996.0267. [DOI] [PubMed] [Google Scholar]
  25. Lamb T. M., Knecht A. K., Smith W. C., Stachel S. E., Economides A. N., Stahl N., Yancopolous G. D., Harland R. M. Neural induction by the secreted polypeptide noggin. Science. 1993 Oct 29;262(5134):713–718. doi: 10.1126/science.8235591. [DOI] [PubMed] [Google Scholar]
  26. Lee J. E., Hollenberg S. M., Snider L., Turner D. L., Lipnick N., Weintraub H. Conversion of Xenopus ectoderm into neurons by NeuroD, a basic helix-loop-helix protein. Science. 1995 May 12;268(5212):836–844. doi: 10.1126/science.7754368. [DOI] [PubMed] [Google Scholar]
  27. Li L., Zhou J., James G., Heller-Harrison R., Czech M. P., Olson E. N. FGF inactivates myogenic helix-loop-helix proteins through phosphorylation of a conserved protein kinase C site in their DNA-binding domains. Cell. 1992 Dec 24;71(7):1181–1194. doi: 10.1016/s0092-8674(05)80066-2. [DOI] [PubMed] [Google Scholar]
  28. Naya F. J., Stellrecht C. M., Tsai M. J. Tissue-specific regulation of the insulin gene by a novel basic helix-loop-helix transcription factor. Genes Dev. 1995 Apr 15;9(8):1009–1019. doi: 10.1101/gad.9.8.1009. [DOI] [PubMed] [Google Scholar]
  29. Ruiz i Altaba A., Cox C., Jessell T. M., Klar A. Ectopic neural expression of a floor plate marker in frog embryos injected with the midline transcription factor Pintallavis. Proc Natl Acad Sci U S A. 1993 Sep 1;90(17):8268–8272. doi: 10.1073/pnas.90.17.8268. [DOI] [PMC free article] [PubMed] [Google Scholar]
  30. Saha M. S., Grainger R. M. Early opsin expression in Xenopus embryos precedes photoreceptor differentiation. Brain Res Mol Brain Res. 1993 Mar;17(3-4):307–318. doi: 10.1016/0169-328x(93)90016-i. [DOI] [PubMed] [Google Scholar]
  31. Sasai Y., Kageyama R., Tagawa Y., Shigemoto R., Nakanishi S. Two mammalian helix-loop-helix factors structurally related to Drosophila hairy and Enhancer of split. Genes Dev. 1992 Dec;6(12B):2620–2634. doi: 10.1101/gad.6.12b.2620. [DOI] [PubMed] [Google Scholar]
  32. Sasai Y., Lu B., Steinbeisser H., De Robertis E. M. Regulation of neural induction by the Chd and Bmp-4 antagonistic patterning signals in Xenopus. Nature. 1995 Jul 27;376(6538):333–336. doi: 10.1038/376333a0. [DOI] [PubMed] [Google Scholar]
  33. Sharpe C. R. Developmental expression of a neurofilament-M and two vimentin-like genes in Xenopus laevis. Development. 1988 Jun;103(2):269–277. doi: 10.1242/dev.103.2.269. [DOI] [PubMed] [Google Scholar]
  34. Sharpe C. R., Pluck A., Gurdon J. B. XIF3, a Xenopus peripherin gene, requires an inductive signal for enhanced expression in anterior neural tissue. Development. 1989 Dec;107(4):701–714. doi: 10.1242/dev.107.4.701. [DOI] [PubMed] [Google Scholar]
  35. Shimizu C., Akazawa C., Nakanishi S., Kageyama R. MATH-2, a mammalian helix-loop-helix factor structurally related to the product of Drosophila proneural gene atonal, is specifically expressed in the nervous system. Eur J Biochem. 1995 Apr 1;229(1):239–248. doi: 10.1111/j.1432-1033.1995.tb20461.x. [DOI] [PubMed] [Google Scholar]
  36. Smith J. C., Price B. M., Green J. B., Weigel D., Herrmann B. G. Expression of a Xenopus homolog of Brachyury (T) is an immediate-early response to mesoderm induction. Cell. 1991 Oct 4;67(1):79–87. doi: 10.1016/0092-8674(91)90573-h. [DOI] [PubMed] [Google Scholar]
  37. Stutz F., Spohr G. Isolation and characterization of sarcomeric actin genes expressed in Xenopus laevis embryos. J Mol Biol. 1986 Feb 5;187(3):349–361. doi: 10.1016/0022-2836(86)90438-9. [DOI] [PubMed] [Google Scholar]
  38. Takebayashi K., Akazawa C., Nakanishi S., Kageyama R. Structure and promoter analysis of the gene encoding the mouse helix-loop-helix factor HES-5. Identification of the neural precursor cell-specific promoter element. J Biol Chem. 1995 Jan 20;270(3):1342–1349. doi: 10.1074/jbc.270.3.1342. [DOI] [PubMed] [Google Scholar]
  39. Takebayashi K., Sasai Y., Sakai Y., Watanabe T., Nakanishi S., Kageyama R. Structure, chromosomal locus, and promoter analysis of the gene encoding the mouse helix-loop-helix factor HES-1. Negative autoregulation through the multiple N box elements. J Biol Chem. 1994 Feb 18;269(7):5150–5156. [PubMed] [Google Scholar]
  40. Tomita K., Ishibashi M., Nakahara K., Ang S. L., Nakanishi S., Guillemot F., Kageyama R. Mammalian hairy and Enhancer of split homolog 1 regulates differentiation of retinal neurons and is essential for eye morphogenesis. Neuron. 1996 Apr;16(4):723–734. doi: 10.1016/s0896-6273(00)80093-8. [DOI] [PubMed] [Google Scholar]
  41. Turner D. L., Weintraub H. Expression of achaete-scute homolog 3 in Xenopus embryos converts ectodermal cells to a neural fate. Genes Dev. 1994 Jun 15;8(12):1434–1447. doi: 10.1101/gad.8.12.1434. [DOI] [PubMed] [Google Scholar]
  42. Walther C., Gruss P. Pax-6, a murine paired box gene, is expressed in the developing CNS. Development. 1991 Dec;113(4):1435–1449. doi: 10.1242/dev.113.4.1435. [DOI] [PubMed] [Google Scholar]
  43. Wright C. V., Morita E. A., Wilkin D. J., De Robertis E. M. The Xenopus XIHbox 6 homeo protein, a marker of posterior neural induction, is expressed in proliferating neurons. Development. 1990 May;109(1):225–234. doi: 10.1242/dev.109.1.225. [DOI] [PubMed] [Google Scholar]
  44. Yasunami M., Suzuki K., Maruyama H., Kawakami H., Nagai Y., Hagiwara M., Ohkubo H. Molecular cloning and characterization of a cDNA encoding a novel basic helix-loop-helix protein structurally related to Neuro-D/BHF1. Biochem Biophys Res Commun. 1996 Mar 27;220(3):754–758. doi: 10.1006/bbrc.1996.0476. [DOI] [PubMed] [Google Scholar]
  45. Zaraisky A. G., Lukyanov S. A., Vasiliev O. L., Smirnov Y. V., Belyavsky A. V., Kazanskaya O. V. A novel homeobox gene expressed in the anterior neural plate of the Xenopus embryo. Dev Biol. 1992 Aug;152(2):373–382. doi: 10.1016/0012-1606(92)90144-6. [DOI] [PubMed] [Google Scholar]
  46. Zimmerman K., Shih J., Bars J., Collazo A., Anderson D. J. XASH-3, a novel Xenopus achaete-scute homolog, provides an early marker of planar neural induction and position along the mediolateral axis of the neural plate. Development. 1993 Sep;119(1):221–232. doi: 10.1242/dev.119.1.221. [DOI] [PubMed] [Google Scholar]

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