The TLX2 Homeobox Gene is a Transcriptional Target of PHOX2B in Neural-crest-derived Cells

Overview

Journal Biochem J

Specialty Biochemistry

Date 2006 Jan 13

PMID 16402914

Citations 21

Authors

Silvia Borghini

Tiziana Bachetti

Monica Fava

Marco Di Duca

Francesca Cargnin

Diego Fornasari

Roberto Ravazzolo

Isabella Ceccherini

Affiliations

Soon will be listed here.

Abstract

The TLX2 (HOX11L1, Ncx, Enx) and PHOX2B genes encode transcription factors crucial in the development of neural-crest-derived cells, leading to ANS (autonomic nervous system) specific neuronal lineages. Moreover, they share a similar expression pattern and are both involved in downstream steps of BMP (bone morphogenetic protein) signalling. In an attempt to reconstruct the gene network sustaining the correct development of the ANS, we have undertaken an in vitro experimental strategy to identify direct upstream regulators of the TLX2 gene. After characterizing a sequence displaying enhancer property in its 5' flanking region, we confirmed the functional link between the human PHOX2B and TLX2 genes. Transient transfections and electrophoretic-mobility-shift assays suggested that PHOX2B is able to bind the cell-specific element in the 5' regulatory region of the TLX2 gene, determining its transactivation in neuroblastoma cells. Such interaction was also confirmed in vivo by means of chromatin immunoprecipitation assay and, in addition, up-regulation of endogenous TLX2 mRNA level was demonstrated following PHOX2B over-expression, by quantitative real-time PCR. Finally, PHOX2B proteins carrying mutations responsible for CCHS (congenital central hypoventilation syndrome) development showed a severe impairment in activating TLX2 expression, both in vitro and in vivo. Taken together, these results support the PHOX2B-TLX2 promoter interaction, suggesting a physiological role in the transcription-factor cascade underlying the differentiation of neuronal lineages of the ANS during human embryogenesis.

Citing Articles

Meta-atlas of Juvenile and Adult Enteric Neuron scRNA-seq for Dataset Comparisons and Consensus on Transcriptomic Definitions of Enteric Neuron Subtypes.

Benthal J, May-Zhang A, Southard-Smith E bioRxiv. 2024; .

PMID: 39574584 PMC: 11580969. DOI: 10.1101/2024.10.31.621315.

Identification of a histone deacetylase inhibitor as a therapeutic candidate for congenital central hypoventilation syndrome.

Africano C, Bachetti T, Uva P, Pitollat G, Del Zotto G, Giacopelli F Mol Ther Nucleic Acids. 2024; 35(4):102319.

PMID: 39329148 PMC: 11426119. DOI: 10.1016/j.omtn.2024.102319.

Comprehensive analysis of TLX2 in pan cancer as a prognostic and immunologic biomarker and validation in ovarian cancer.

Chen B, Ding X, Wan A, Qi X, Lin X, Wang H Sci Rep. 2023; 13(1):16244.

PMID: 37758722 PMC: 10533500. DOI: 10.1038/s41598-023-42171-5.

Unbiased characterization of the larval zebrafish enteric nervous system at a single cell transcriptomic level.

Kuil L, Kakiailatu N, Windster J, Bindels E, Zink J, van der Zee G iScience. 2023; 26(7):107070.

PMID: 37426341 PMC: 10329177. DOI: 10.1016/j.isci.2023.107070.

Sulforaphane improves mitochondrial metabolism in fibroblasts from patients with fragile X-associated tremor and ataxia syndrome.

Napoli E, Flores A, Mansuri Y, Hagerman R, Giulivi C Neurobiol Dis. 2021; 157:105427.

PMID: 34153466 PMC: 8475276. DOI: 10.1016/j.nbd.2021.105427.

References

Costa M, Fava M, Seri M, Cusano R, Sancandi M, Forabosco P . Evaluation of the HOX11L1 gene as a candidate for congenital disorders of intestinal innervation. J Med Genet. 2000; 37(7):E9. PMC: 1734627. DOI: 10.1136/jmg.37.7.e9. View

Shirasawa S, Yunker A, Roth K, Brown G, Horning S, Korsmeyer S . Enx (Hox11L1)-deficient mice develop myenteric neuronal hyperplasia and megacolon. Nat Med. 1997; 3(6):646-50. DOI: 10.1038/nm0697-646. View

Amiel J, Laudier B, Attie-Bitach T, Trang H, De Pontual L, Gener B . Polyalanine expansion and frameshift mutations of the paired-like homeobox gene PHOX2B in congenital central hypoventilation syndrome. Nat Genet. 2003; 33(4):459-61. DOI: 10.1038/ng1130. View

Parisi M, Baldessari A, Iida M, Clarke C, Doggett B, Shirasawa S . Genetic background modifies intestinal pseudo-obstruction and the expression of a reporter gene in Hox11L1-/- mice. Gastroenterology. 2003; 125(5):1428-40. DOI: 10.1016/j.gastro.2003.08.021. View

Puri P . Intestinal neuronal dysplasia. Semin Pediatr Surg. 2003; 12(4):259-64. DOI: 10.1053/j.sempedsurg.2003.08.007. View

Matera I, Bachetti T, Puppo F, Di Duca M, Morandi F, Casiraghi G . PHOX2B mutations and polyalanine expansions correlate with the severity of the respiratory phenotype and associated symptoms in both congenital and late onset Central Hypoventilation syndrome. J Med Genet. 2004; 41(5):373-80. PMC: 1735781. DOI: 10.1136/jmg.2003.015412. View

Yanai T, Kobayashi H, Yamataka A, Lane G, Miyano T, Hayakawa T . Acetylcholine-related bowel dysmotility in homozygous mutant NCX/HOX11L.1-deficient (NCX-/-) mice-evidence that acetylcholine is implicated in causing intestinal neuronal dysplasia. J Pediatr Surg. 2004; 39(6):927-30. DOI: 10.1016/j.jpedsurg.2004.02.004. View

Dignam J, Lebovitz R, Roeder R . Accurate transcription initiation by RNA polymerase II in a soluble extract from isolated mammalian nuclei. Nucleic Acids Res. 1983; 11(5):1475-89. PMC: 325809. DOI: 10.1093/nar/11.5.1475. View

Higuchi R, Krummel B, Saiki R . A general method of in vitro preparation and specific mutagenesis of DNA fragments: study of protein and DNA interactions. Nucleic Acids Res. 1988; 16(15):7351-67. PMC: 338413. DOI: 10.1093/nar/16.15.7351. View

10.

Roberts C, Sonder A, Lumsden A, Korsmeyer S . Development expression of Hox11 and specification of splenic cell fate. Am J Pathol. 1995; 146(5):1089-101. PMC: 1869297. View

11.

Quandt K, Frech K, Karas H, Wingender E, Werner T . MatInd and MatInspector: new fast and versatile tools for detection of consensus matches in nucleotide sequence data. Nucleic Acids Res. 1995; 23(23):4878-84. PMC: 307478. DOI: 10.1093/nar/23.23.4878. View

12.

Hatano M, Iitsuka Y, Yamamoto H, Dezawa M, Yusa S, Kohno Y . Ncx, a Hox11 related gene, is expressed in a variety of tissues derived from neural crest cells. Anat Embryol (Berl). 1997; 195(5):419-25. DOI: 10.1007/s004290050061. View

13.

Hatano M, Aoki T, Dezawa M, Yusa S, Iitsuka Y, Koseki H . A novel pathogenesis of megacolon in Ncx/Hox11L.1 deficient mice. J Clin Invest. 1997; 100(4):795-801. PMC: 508250. DOI: 10.1172/JCI119593. View

14.

Pattyn A, Morin X, Cremer H, Goridis C, Brunet J . Expression and interactions of the two closely related homeobox genes Phox2a and Phox2b during neurogenesis. Development. 1997; 124(20):4065-75. DOI: 10.1242/dev.124.20.4065. View

15.

Tang S, Hoodless P, Lu Z, Breitman M, McInnes R, Wrana J . The Tlx-2 homeobox gene is a downstream target of BMP signalling and is required for mouse mesoderm development. Development. 1998; 125(10):1877-87. DOI: 10.1242/dev.125.10.1877. View

16.

Kim H, Seo H, Yang C, Brunet J, Kim K . Noradrenergic-specific transcription of the dopamine beta-hydroxylase gene requires synergy of multiple cis-acting elements including at least two Phox2a-binding sites. J Neurosci. 1998; 18(20):8247-60. PMC: 6792837. View

17.

Yang C, Kim H, Seo H, Kim C, Brunet J, Kim K . Paired-like homeodomain proteins, Phox2a and Phox2b, are responsible for noradrenergic cell-specific transcription of the dopamine beta-hydroxylase gene. J Neurochem. 1998; 71(5):1813-26. DOI: 10.1046/j.1471-4159.1998.71051813.x. View

18.

Lo L, Morin X, Brunet J, Anderson D . Specification of neurotransmitter identity by Phox2 proteins in neural crest stem cells. Neuron. 1999; 22(4):693-705. DOI: 10.1016/s0896-6273(00)80729-1. View

19.

Pattyn A, Morin X, Cremer H, Goridis C, Brunet J . The homeobox gene Phox2b is essential for the development of autonomic neural crest derivatives. Nature. 1999; 399(6734):366-70. DOI: 10.1038/20700. View

20.

Iitsuka Y, Shimizu H, Kang M, Sasagawa K, Sekiya S, Tokuhisa T . An enhancer element for expression of the Ncx (Enx, Hox11L1) gene in neural crest-derived cells. J Biol Chem. 1999; 274(34):24401-7. DOI: 10.1074/jbc.274.34.24401. View