Mammalian Numb-interacting Protein 1/Dual Oxidase Maturation Factor 1 Directs Neuronal Fate in Stem Cells

Authors

Karen A. M. Kennedy, The University of Western Ontario
Elena A. Ostrakhovitch, The University of Western Ontario
Shelley D. E. Sandiford, The University of Western Ontario
Thamara Dayarathna, The University of Western Ontario
Xiaojun Xie, The University of Western Ontario
Elaine Y. L. Waese, University of Toronto
Wing Y. Chang, University of Toronto
Qingping Feng, The University of Western Ontario
Ilona S. Skerjanc, The University of Western Ontario
William L. Stanford, University of Toronto
Shawn S. C. Li, The University of Western Ontario

Document Type

Article

Publication Date

6-4-2010

Journal

Journal of Biological Chemistry

Volume

285

Issue

23

First Page

17974

Last Page

17985

URL with Digital Object Identifier

http://dx.doi.org/10.1074/jbc.M109.084616

Abstract

In this study, we describe a role for the mammalian Numb-interacting protein 1 (Nip1) in regulation of neuronal differentiation in stem cells. The expression of Nip1 was detected in the developing mouse brain, embryonic stem cells, primary neuronal stem cells, and retinoic acid-treated P19 embryonal carcinoma cells. The highest expression of Nip1 was observed in undifferentiated neuronal stem cells and was associated with Duox1-mediated reactive oxygen species ROS production. Ectopic nip1 expression in P19 embryonal carcinoma cells induced neuronal differentiation, and this phenotype was also linked to elevated ROS production. The neuronal differentiation in nip1-overexpressing P19 cells was achieved in a retinoic acid-independent manner and was corroborated by an increase in the expression of the neuronal basic helix-loop-helix transcription factors and neural-lineage cell markers. Furthermore, depletion of nip1 by short hairpin RNA led to a decrease in the expression of neuronal basic helix-loop-helix transcription factors and ROS. However, inhibition of ROS production in nip1-overexpressing P19 cells restricted but did not extinguish neuronal differentiation. Microarray and mass spectrometry analysis identified intermediate filaments as the principal cytoskeletal elements affected by up-regulation of nip1. We show here the first evidence for a functional interaction between Nip1 and a component of the nuclear lamina, lamin A/C. associated with a neuronal-specific phenotype. Taken together, our data reveal an important role for Nip1 in the guidance of neuronal differentiation through ROS generation and modulation of intermediate filaments and implicate Nip1 as a novel intrinsic regulator of neuronal cell fate.