Intrinsic HER4/4ICD transcriptional activation domains are required for STAT5A activated gene expression

Gene. 2016 Oct 30;592(1):221-226. doi: 10.1016/j.gene.2016.07.071. Epub 2016 Aug 5.


The epidermal growth factor receptor family member HER4 undergoes proteolytic processing at the cell surface to release the HER4 intracellular domain (4ICD) nuclear protein. Interestingly, 4ICD directly interacts with STAT5 and functions as an obligate STAT5 nuclear chaperone. Once in the nucleus 4ICD binds with STAT5 at STAT5 target genes, dramatically potentiating STAT5 transcriptional activation. These observations raise the possibility that 4ICD directly coactivates STAT5 gene expression. Using both yeast and mammalian transactivation reporter assays, we performed truncations of 4ICD fused to a GAL4 DNA binding domain and identified two independent 4ICD transactivation domains located between residues 1022 and 1090 (TAD1) and 1192 and 1225 (TAD2). The ability of the 4ICD DNA binding domain fusions to transactivate reporter gene expression required deletion of the intrinsic tyrosine kinase domain. In addition, we identified the 4ICD carboxyl terminal TVV residues, a PDZ domain binding motif (PDZ-DBM), as a potent transcriptional repressor. The transactivation activity of the HER4 carboxyl terminal domain lacking the tyrosine kinase (CTD) was significantly lower than similar EGFR or HER2 CTD. However, deletion of the HER4 CTD PDZ-DBM enhanced HER4 CTD transactivation to levels equivalent to the EGFR and HER2 CTDs. To determine if 4ICD TAD1 and TAD2 have a physiologically relevant role in STAT5 transactivation, we coexpressed 4ICD or 4ICD lacking TAD2 or both TAD1 and TAD2 with STAT5 in a luciferase reporter assay. Our results demonstrate that each 4ICD TAD contributes additively to STAT5A transactivation and the ability of STAT5A to transactivate the β-casein promoter requires the 4ICD TADs. Taken together, published data and our current results demonstrate that both 4ICD nuclear chaperone and intrinsic coactivation activities are essential for STAT5 regulated gene expression.

Keywords: 4ICD; EGFR-family; HER4; Regulated intramembrane proteolysis; STAT5; Transactivation.

MeSH terms

  • Binding Sites
  • HEK293 Cells
  • Humans
  • MCF-7 Cells
  • Protein Binding
  • Receptor, ErbB-4 / chemistry
  • Receptor, ErbB-4 / genetics
  • Receptor, ErbB-4 / metabolism*
  • STAT5 Transcription Factor / chemistry
  • STAT5 Transcription Factor / metabolism*
  • Transcriptional Activation*
  • Tumor Suppressor Proteins / chemistry
  • Tumor Suppressor Proteins / metabolism*


  • STAT5 Transcription Factor
  • STAT5A protein, human
  • Tumor Suppressor Proteins
  • ERBB4 protein, human
  • Receptor, ErbB-4