Inactivation of IκB contributes to transcriptional activation of spermidine/spermine N(1)-acetyltransferase

Woonyoung Choi, Lynsey Proctor, Qianghua Xia, Yumei Feng, Eugene W. Gerner, Paul J. Chiao, Stanley R. Hamilton, Wei Zhang

Research output: Contribution to journalArticlepeer-review

Abstract

Spermidine/spermine N(1)-acetyltransferase (SSAT) is a key enzyme in polyamine catabolism. We recently reported that the combination of N 1, N11-diethylnorspermine (DENSPM) and 5-fluorouracil (5-FU) synergistically induces SSAT expression, depletes polyamine levels and causes apoptosis in colon cancer cells. To determine whether new RNA and protein synthesis is required for SSAT induction, we examined the effect of actinomycin D (ActD) and cycloheximide (CHX). ActD alone blocked the induction of SSAT expression; however, the combination of CHX and DENSPM markedly induced SSAT expression and caused mitochondrial damage, suggesting that an inhibitory labile protein is involved in SSAT transactivation. SSAT promoter analysis identified two putative Rel/Nuclear Factor κB (NFκB) binding sites. Thus, we hypothesized that IκB is the labile inhibitory protein and that its removal contributes to the activation of NFκB. CHX quickly eliminated the IκB protein in the cells and increased the levels of the two subunits of NFκB, p65 and p50, in the nucleus. Luciferase reporter gene assay showed that SSAT promoter constructs containing the two putative NFκB binding elements responded to CHX as well as TNFα, whereas the promoter without the two sites did not. Chromatin immunoprecipitation (ChIP) assay showed that NFκB was indeed bound to the SSAT promoter after CHX treatment. Further, dominant negative IκB attenuated the CHX and DENSPM-induced SSAT expression and mitochondria damage. These results taken together suggest that the inhibition of IκB and activation of NFκB activate SSAT.

Original languageEnglish (US)
Pages (from-to)685-693
Number of pages9
JournalMolecular Carcinogenesis
Volume45
Issue number9
DOIs
StatePublished - Sep 2006
Externally publishedYes

Keywords

  • Cycloheximide
  • Labile protein
  • N
  • N-diethylnorspermine

ASJC Scopus subject areas

  • Molecular Biology
  • Cancer Research

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