Physical and functional interactions between the transactivation domain of the hematopoietic transcription factor NF-E2 and WW domains

Eric A. Mosser, Jeremy D. Kasanov, E. Camilla Forsberg, Brian K. Kay, Paul A. Ney, Emery H. Bresnick*

*Corresponding author for this work

Research output: Contribution to journalArticlepeer-review

60 Scopus citations

Abstract

Tandem binding sites for the hematopoietic transcription factor NF-E2 in the β-globin locus control region activate high-level β-globin gene expression in transgenic mice. NF-E2 is a heterodimer consisting of a hematopoietic subunit p45 and a ubiquitous subunit p18. Gavva et al. [Gavva, N. R, Gavva, R., Ermekova, K., Sudol, M., and Shen, J. C. (1997) J. Biol. Chem. 272, 24105-24108] reported that human p45 contains a PPXY motif that binds WW domains. We show that murine NF-E2, which contains two PPXY motifs (PPXY-1 and -2) within its transactivation domain, differentially interacted with nine GST-WW domain fusion proteins. Quantitative analysis revealed high- affinity binding (K(D) = 5.7 nM) of p45 to a WW domain from a novel human ubiquitin ligase homologue (WWP1) expressed in hematopoietic tissues. The amino-terminal WW domain of WWP1 formed a multimeric complex with DNA-bound NF-E2. A WWP1 ligand peptide, isolated by phage display, and a peptide spanning PPXY-1 inhibited p45 binding, whereas an SH3 domain-interacting peptide and a peptide spanning PPXY-2 did not. Mutation of PPXY-1, but not PPXY-2, inhibited the transactivation function of NF-E2, providing support for the hypothesis that WW domain interactions are important for NF-E2- mediated transactivation.

Original languageEnglish (US)
Pages (from-to)13686-13695
Number of pages10
JournalBiochemistry
Volume37
Issue number39
DOIs
StatePublished - Sep 29 1998

ASJC Scopus subject areas

  • Biochemistry

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