DRIP150 coactivation of estrogen receptor α in ZR-75 breast cancer cells is independent of LXXLL motifs

Eun Lee Jeongeun, Kyounghyun Kim, James C. Sacchettini, Clare V. Smith, Stephen Safe

Research output: Contribution to journalArticle

18 Scopus citations

Abstract

Vitamin D receptor-interacting protein 150 (DRIP150) has been identified as part of mediator-like complexes that enhance transcriptional activation of the estrogen receptor (ER) and other nuclear receptors (NRs). DRIP150 coactivates ligand-dependent ERα-mediated transactivation in ZR-75 and MDA-MB-231 breast cancer cells transfected with a (luciferase) reporter construct (pERE 3) regulated by three tandem estrogen-responsive elements. Coactivation of ERα by DRIP150 in ZR-75 cells was activation function 2-dependent and required an intact helix 12 that typically interacts with LXXLL motifs (NR box) in p160 steroid receptor coactivators. DRIP150 contains C- and N-terminal NR boxes (amino acids 1182-1186 and 69-73, respectively), and deletion analysis of DRIP150 showed that regions containing these sequences were not necessary for coactivation of ERα. Analysis of multiple DRIP150 deletion mutants identified a 23-amino-acid sequence (789-811) required for coactivation activity. Analysis of the protein crystal structure data base identified two regions at amino acids 789-794 and 795-804, which resembled α-helical motifs in Lanuginosa lipase/histamine N-methyltransferase and hepatocyte nuclear factor 1, respectively. By using a squelching assay and specific amino acid point mutations within each α-helix, the NIFSEVRVYN (795-804) region was identified as the critical sequence required for the activity of DRIP150. These results demonstrate that coactivati'on of ERa by DRIP150 in ZR-75 cells is NR box-independent and requires a novel sequence with putative α-helical structure.

Original languageEnglish (US)
Pages (from-to)8819-8830
Number of pages12
JournalJournal of Biological Chemistry
Volume280
Issue number10
DOIs
StatePublished - Mar 11 2005

ASJC Scopus subject areas

  • Biochemistry
  • Molecular Biology
  • Cell Biology

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