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. 2016 Jun 21;2:16011. doi: 10.1038/celldisc.2016.11

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Copyright © 2016 SIBS, CAS

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Arrdc1 and Arrdc4 act as Nedd4 ubiquitin ligase adaptors to regulate DMT1. (A) The relative transport activity of endogenous DMT1 measured using the fluorescence quenching assay is regulated by Arrdc1 and Arrdc4 in a PY-dependent manner in Caco-2 cells. Data are mean±s.e.m., n=3–6. ***P<0.001, *P<0.05. Arrdc1 (B) and Arrdc4 (C) interact with DMT1, and this interaction is not dependent on the PY motif. (D) The plasma membrane localization of DMT1 decreases with Arrdc1 and Arrdc4 and is dependent on its PY motif. When Arrdc1 (E) or Arrdc4 (F) is present, DMT1 levels are not stabilized, even after MG132 (proteosomal inhibitor) and chloroquine (CHQ; lysosomal inhibitor) treatment. (G) DMT1 ubiquitination by Nedd4-2 decreases in the presence of Arrdc1 or Arrdc4.