Transglutaminase-catalyzed incorporation of polyamines masks the DNA-binding region of the transcription factor Relish

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Abstract

In Drosophila, the final immune deficiency (IMD) pathwaydependent signal is transmitted through proteolytic conversion of the nuclear factor-B (NF-B)-like transcription factor Relish to the active N-Terminal fragment Relish-N. Relish-N is then translocated from the cytosol into the nucleus for the expression of IMD-controlled genes. We previously demonstrated that transglutaminase (TG) suppresses the IMD pathway by polymerizing Relish-N to inhibit its nuclear translocation. Conversely, we also demonstrated that orally ingested synthetic amines, such as monodansylcadaverine (DCA) and biotin-labeled pentylamine, are TG-dependently incorporated into Relish-N, causing the nuclear translocation of modified Relish-N in gut epithelial cells. It remains unclear, however, whether polyamine- containing Relish-N retains transcriptional activity. Here, we used mass spectrometry analysis of a recombinant Relish- N modified with DCA by TG activity after proteolytic digestion and show that the DCA-modified Gln residues are located in the DNA-binding region of Relish-N. TG-catalyzed DCA incorporation inhibited binding of Relish-N to the Rel-responsive element in the NF-B-binding DNA sequence. Subcellular fractionation of TG-expressing Drosophila S2 cells indicated that TG was localized in both the cytosol and nucleus. Of note, natural polyamines, including spermidine and spermine, competitively inhibited TG-dependentDCAincorporation into Relish- N. Moreover, in vivo experiments demonstrated that Relish- N was modified by spermine and that this modification reduced transcription of IMD pathway-controlled cecropin A1 and diptericin genes. These findings suggest that intracellular TG regulates Relish-N-mediated transcriptional activity by incorporating polyamines into Relish-N and via protein-protein cross-linking.

Original languageEnglish
Pages (from-to)6369-6380
Number of pages12
JournalJournal of Biological Chemistry
Volume292
Issue number15
DOIs
Publication statusPublished - Apr 14 2017

All Science Journal Classification (ASJC) codes

  • Biochemistry
  • Molecular Biology
  • Cell Biology

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