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. 1993 Oct 1;90(19):9120–9124. doi: 10.1073/pnas.90.19.9120

Neurotrophic activity of the Antennapedia homeodomain depends on its specific DNA-binding properties.

I Le Roux 1, A H Joliot 1, E Bloch-Gallego 1, A Prochiantz 1, M Volovitch 1
PMCID: PMC47513  PMID: 8105471

Abstract

In previous reports we have demonstrated that the 60-aa peptide corresponding to the homeodomain of the Drosophila protein Antennapedia (pAntp) translocates through the membrane of neurons in culture, accumulates in neuronal nuclei, and promotes neurite growth. To analyze the importance of specific pAntp DNA-binding properties in this phenomenon we have constructed three mutant versions of pAntp that differ in their ability to translocate through the membrane and to bind specifically the cognate sequence for homeodomains present in the promoter of HoxA5. We demonstrate that removing two hydrophobic residues of the third helix inhibits pAntp internalization and suppresses its neurotrophic activity. We also show that pAntp neurotrophic activity is lost when mutations are introduced in positions preserving its penetration and nuclear accumulation but abolishing its capacity to bind specifically the cognate DNA-binding motif for homeoproteins. Our results strongly suggest that pAntp neurotrophicity requires both its internalization and its specific binding to homeobox cognate sequences. We propose that homeoproteins might regulate important events in the morphological differentiation of the postmitotic neuron.

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Selected References

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

  1. Bloch-Gallego E., Le Roux I., Joliot A. H., Volovitch M., Henderson C. E., Prochiantz A. Antennapedia homeobox peptide enhances growth and branching of embryonic chicken motoneurons in vitro. J Cell Biol. 1993 Jan;120(2):485–492. doi: 10.1083/jcb.120.2.485. [DOI] [PMC free article] [PubMed] [Google Scholar]
  2. Chen E. Y., Seeburg P. H. Supercoil sequencing: a fast and simple method for sequencing plasmid DNA. DNA. 1985 Apr;4(2):165–170. doi: 10.1089/dna.1985.4.165. [DOI] [PubMed] [Google Scholar]
  3. Chisaka O., Musci T. S., Capecchi M. R. Developmental defects of the ear, cranial nerves and hindbrain resulting from targeted disruption of the mouse homeobox gene Hox-1.6. Nature. 1992 Feb 6;355(6360):516–520. doi: 10.1038/355516a0. [DOI] [PubMed] [Google Scholar]
  4. Cho K. W., Morita E. A., Wright C. V., De Robertis E. M. Overexpression of a homeodomain protein confers axis-forming activity to uncommitted Xenopus embryonic cells. Cell. 1991 Apr 5;65(1):55–64. doi: 10.1016/0092-8674(91)90407-p. [DOI] [PubMed] [Google Scholar]
  5. Doe C. Q., Hiromi Y., Gehring W. J., Goodman C. S. Expression and function of the segmentation gene fushi tarazu during Drosophila neurogenesis. Science. 1988 Jan 8;239(4836):170–175. doi: 10.1126/science.2892267. [DOI] [PubMed] [Google Scholar]
  6. Hanes S. D., Brent R. DNA specificity of the bicoid activator protein is determined by homeodomain recognition helix residue 9. Cell. 1989 Jun 30;57(7):1275–1283. doi: 10.1016/0092-8674(89)90063-9. [DOI] [PubMed] [Google Scholar]
  7. Joliot A. H., Triller A., Volovitch M., Pernelle C., Prochiantz A. alpha-2,8-Polysialic acid is the neuronal surface receptor of antennapedia homeobox peptide. New Biol. 1991 Nov;3(11):1121–1134. [PubMed] [Google Scholar]
  8. Joliot A., Pernelle C., Deagostini-Bazin H., Prochiantz A. Antennapedia homeobox peptide regulates neural morphogenesis. Proc Natl Acad Sci U S A. 1991 Mar 1;88(5):1864–1868. doi: 10.1073/pnas.88.5.1864. [DOI] [PMC free article] [PubMed] [Google Scholar]
  9. Jones F. S., Chalepakis G., Gruss P., Edelman G. M. Activation of the cytotactin promoter by the homeobox-containing gene Evx-1. Proc Natl Acad Sci U S A. 1992 Mar 15;89(6):2091–2095. doi: 10.1073/pnas.89.6.2091. [DOI] [PMC free article] [PubMed] [Google Scholar]
  10. Jones F. S., Prediger E. A., Bittner D. A., De Robertis E. M., Edelman G. M. Cell adhesion molecules as targets for Hox genes: neural cell adhesion molecule promoter activity is modulated by cotransfection with Hox-2.5 and -2.4. Proc Natl Acad Sci U S A. 1992 Mar 15;89(6):2086–2090. doi: 10.1073/pnas.89.6.2086. [DOI] [PMC free article] [PubMed] [Google Scholar]
  11. Lafont F., Rouget M., Triller A., Prochiantz A., Rousselet A. In vitro control of neuronal polarity by glycosaminoglycans. Development. 1992 Jan;114(1):17–29. doi: 10.1242/dev.114.1.17. [DOI] [PubMed] [Google Scholar]
  12. Le Mouellic H., Lallemand Y., Brûlet P. Homeosis in the mouse induced by a null mutation in the Hox-3.1 gene. Cell. 1992 Apr 17;69(2):251–264. doi: 10.1016/0092-8674(92)90406-3. [DOI] [PubMed] [Google Scholar]
  13. Lufkin T., Dierich A., LeMeur M., Mark M., Chambon P. Disruption of the Hox-1.6 homeobox gene results in defects in a region corresponding to its rostral domain of expression. Cell. 1991 Sep 20;66(6):1105–1119. doi: 10.1016/0092-8674(91)90034-v. [DOI] [PubMed] [Google Scholar]
  14. Martinez S., Wassef M., Alvarado-Mallart R. M. Induction of a mesencephalic phenotype in the 2-day-old chick prosencephalon is preceded by the early expression of the homeobox gene en. Neuron. 1991 Jun;6(6):971–981. doi: 10.1016/0896-6273(91)90237-t. [DOI] [PubMed] [Google Scholar]
  15. McGinnis W., Krumlauf R. Homeobox genes and axial patterning. Cell. 1992 Jan 24;68(2):283–302. doi: 10.1016/0092-8674(92)90471-n. [DOI] [PubMed] [Google Scholar]
  16. McMahon A. P., Joyner A. L., Bradley A., McMahon J. A. The midbrain-hindbrain phenotype of Wnt-1-/Wnt-1- mice results from stepwise deletion of engrailed-expressing cells by 9.5 days postcoitum. Cell. 1992 May 15;69(4):581–595. doi: 10.1016/0092-8674(92)90222-x. [DOI] [PubMed] [Google Scholar]
  17. Miller D. M., Shen M. M., Shamu C. E., Bürglin T. R., Ruvkun G., Dubois M. L., Ghee M., Wilson L. C. elegans unc-4 gene encodes a homeodomain protein that determines the pattern of synaptic input to specific motor neurons. Nature. 1992 Feb 27;355(6363):841–845. doi: 10.1038/355841a0. [DOI] [PubMed] [Google Scholar]
  18. Niehrs C., De Robertis E. M. Ectopic expression of a homeobox gene changes cell fate in Xenopus embryos in a position-specific manner. EMBO J. 1991 Dec;10(12):3621–3629. doi: 10.1002/j.1460-2075.1991.tb04928.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  19. Odenwald W. F., Garbern J., Arnheiter H., Tournier-Lasserve E., Lazzarini R. A. The Hox-1.3 homeo box protein is a sequence-specific DNA-binding phosphoprotein. Genes Dev. 1989 Feb;3(2):158–172. doi: 10.1101/gad.3.2.158. [DOI] [PubMed] [Google Scholar]
  20. Perez F., Joliot A., Bloch-Gallego E., Zahraoui A., Triller A., Prochiantz A. Antennapedia homeobox as a signal for the cellular internalization and nuclear addressing of a small exogenous peptide. J Cell Sci. 1992 Aug;102(Pt 4):717–722. doi: 10.1242/jcs.102.4.717. [DOI] [PubMed] [Google Scholar]
  21. Rosenberg A. H., Lade B. N., Chui D. S., Lin S. W., Dunn J. J., Studier F. W. Vectors for selective expression of cloned DNAs by T7 RNA polymerase. Gene. 1987;56(1):125–135. doi: 10.1016/0378-1119(87)90165-x. [DOI] [PubMed] [Google Scholar]
  22. Treisman J., Gönczy P., Vashishtha M., Harris E., Desplan C. A single amino acid can determine the DNA binding specificity of homeodomain proteins. Cell. 1989 Nov 3;59(3):553–562. doi: 10.1016/0092-8674(89)90038-x. [DOI] [PubMed] [Google Scholar]
  23. Violette S. M., Shashikant C. S., Salbaum J. M., Belting H. G., Wang J. C., Ruddle F. H. Repression of the beta-amyloid gene in a Hox-3.1-producing cell line. Proc Natl Acad Sci U S A. 1992 May 1;89(9):3805–3809. doi: 10.1073/pnas.89.9.3805. [DOI] [PMC free article] [PubMed] [Google Scholar]
  24. Wagner-Bernholz J. T., Wilson C., Gibson G., Schuh R., Gehring W. J. Identification of target genes of the homeotic gene Antennapedia by enhancer detection. Genes Dev. 1991 Dec;5(12B):2467–2480. doi: 10.1101/gad.5.12b.2467. [DOI] [PubMed] [Google Scholar]
  25. White J. G., Southgate E., Thomson J. N. Mutations in the Caenorhabditis elegans unc-4 gene alter the synaptic input to ventral cord motor neurons. Nature. 1992 Feb 27;355(6363):838–841. doi: 10.1038/355838a0. [DOI] [PubMed] [Google Scholar]

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