In vivo interactions of TTDA mutant proteins within TFIIH

J Cell Sci. 2013 Aug 1;126(Pt 15):3278-83. doi: 10.1242/jcs.126839. Epub 2013 May 31.

Abstract

Trichothiodystrophy group A (TTD-A) patients carry a mutation in the transcription factor II H (TFIIH) subunit TTDA. Using a novel in vivo tripartite split-GFP system, we show that TTDA interacts with the TFIIH subunit p52 and the p52-TTDA-GFP product is incorporated into TFIIH. p52-TTDA-GFP is able to bind DNA and is recruited to UV-damaged DNA. Furthermore, we show that two patient-mutated TTDA proteins can interact with p52, are able to bind to the DNA and can localize to damaged DNA. Our findings give new insights into the behavior of TTDA within the context of a living cell and thereby shed light on the complex phenotype of TTD-A patients.

Keywords: Protein–protein interactions; TFIIH; TTDA; Trichothiodystrophy; Tripartite split-GFP.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Cell Line
  • DNA / metabolism
  • DNA Damage
  • Fibroblasts
  • Humans
  • Mutant Proteins / genetics
  • Mutant Proteins / metabolism
  • Protein Interaction Domains and Motifs
  • Protein Subunits
  • Shc Signaling Adaptor Proteins / genetics
  • Shc Signaling Adaptor Proteins / metabolism
  • Src Homology 2 Domain-Containing, Transforming Protein 1
  • Transcription Factor TFIIH / genetics
  • Transcription Factor TFIIH / metabolism*
  • Transcription Factors / genetics
  • Transcription Factors / metabolism*
  • Transfection
  • Trichothiodystrophy Syndromes / genetics*
  • Trichothiodystrophy Syndromes / metabolism*

Substances

  • GTF2H5 protein, human
  • Mutant Proteins
  • Protein Subunits
  • SHC1 protein, human
  • Shc Signaling Adaptor Proteins
  • Src Homology 2 Domain-Containing, Transforming Protein 1
  • Transcription Factors
  • Transcription Factor TFIIH
  • DNA