Functional analysis of TCF4 missense mutations that cause Pitt-Hopkins syndrome

Marc Forrest, Ria M. Chapman, A. Michelle Doyle, Caroline L. Tinsley, Adrian Waite, Derek J. Blake*

*Corresponding author for this work

Research output: Contribution to journalArticlepeer-review

48 Scopus citations

Abstract

Pitt-Hopkins syndrome (PTHS) is a rare developmental disorder associated with severe mental retardation, facial abnormalities, and intermittent hyperventilation. Autosomal dominant PTHS is caused by mutations in the transcription factor 4 (TCF4) gene, whereas NRXN1 and CNTNAP2 mutations are associated with autosomal recessive PTHS. To determine the impact of missense mutations on TCF4 function, we tested a panel of PTHS-associated mutations using a range of quantitative techniques. Mutations in the basic helix-loop-helix (bHLH) domain of TCF4 alter the subnuclear localization of the mutant protein and can attenuate homo- and heterodimer formation in homogenous time-resolved fluorescence (HTRF) assays. By contrast, mutations proximal to the bHLH domain do not alter the location of TCF4 or impair heterodimer formation. In addition, we show that TCF4 can transactivate the NRXN1β and CNTNAP2 promoters in luciferase assays. Here we find variable, context-specific deficits in the ability of the different PTHS-associated TCF4 mutants to transactivate these promoters when coexpressed with different bHLH transcription factors. These data demonstrate that PTHS-associated missense mutations can have multiple effects on the function of the protein, and suggest that TCF4 may modulate the expression of NRXN1 and CNTNAP2 thereby defining a regulatory network in PTHS.

Original languageEnglish (US)
Pages (from-to)1676-1686
Number of pages11
JournalHuman mutation
Volume33
Issue number12
DOIs
StatePublished - Dec 2012
Externally publishedYes

Keywords

  • Mental retardation
  • Missense mutation
  • Pitt-Hopkins syndrome
  • TCF4
  • Transcription factor

ASJC Scopus subject areas

  • Genetics
  • Genetics(clinical)

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