Differential expression and dynamic changes of murine NEDD9 in progenitor cells of diverse tissues

doi:10.1016/j.gep.2008.01.001

Gene Expression Patterns

Volume 8, Issue 4, April 2008, Pages 217-226

https://doi.org/10.1016/j.gep.2008.01.001 Get rights and content

Abstract

NEDD9 is a scaffolding protein in the integrin signaling pathway that is involved in cell adhesion dynamics. Little is known of the cellular localization of NEDD9 expression during embryonic development. In the present study, we have analyzed NEDD9 mRNA expression in the mouse and identified new relevant expression sites. In addition, we have characterized NEDD9 protein expression pattern for the first time in mammals. At E9.5–E10.5, high levels of Nedd9 and the neurogenic transcription factor neurogenin-2 (Ngn2) were found to largely overlap in two discrete domains of the trunk neural tube along its dorso-ventral axis, with Nedd9 extending to more ventral regions of the ventricular zone and Ngn2 differentially expressed in neuronally committed progenitors of the intermediate zone. At encephalic and trunk levels of the neural tube, NEDD9 was present in Sox2⁺ progenitor cell populations mostly generating Ngn2⁺ and/or Nurr1⁺ cells. A sharp down-regulation of NEDD9 expression was found in cells upon lineage commitment, as observed in Nurr1⁺ and Ngn2⁺ mesencephalic dopaminergic and brainstem neuronal progenitors. In other tissues/organs, i.e. prospective heart, retina, olfactory epithelium, gonads, cartilage, gut and pituitary gland, NEDD9 was found to be co-expressed with Sox2, RXR alpha and/or Nurr1-like proteins, suggesting that NEDD9 expression is confined to early progenitors involved in diverse organogenesis and that it may depend on the repertoire and levels of retinoic acid co-receptors expressed by those cells.

Section snippets

Results and discussion

NEDD9 (also known as HEF1 and CasL) is an adaptor protein of the β1-integrin signaling pathway (O’Neill et al., 2000). NEDD9 belongs to the Cas family of proteins (including p130Cas and Efs/Sin) which share a common domain structure and sequence homology but have potentially different functions, suggested by their differential tissue, cellular and sub-cellular localization and posttranslational modifications (Law et al., 1996, Law et al., 1998, O’Neill et al., 2000, Bouton et al., 2001, Fashena

Materials and methods

All experimental procedures were approved by the Swedish National Board for Laboratory Animals (N17/2003 and N293/05).

Acknowledgements

We thank Thomas Edlund for the Sox2 antibody and Erica A. Golemis and Elena Pugacheva for NEDD9 fragment peptide. We acknowledge Claudia Tello and Jonny Söderlund for technical assistance. This work was supported by the Swedish Medical Research Council and the Swedish Foundation for Strategic Research (CEDB grant, DBRM grant), and support to DBRM. J.B.A. was supported by European Union (Marie Curie RTN-CT-2003-504636).

References (43)

C. Anneren et al.
GTK, a Src-related tyrosine kinase, induces nerve growth factor-independent neurite outgrowth in PC12 cells through activation of the Rap1 pathway. Relationship to Shb tyrosine phosphorylation and elevated levels of focal adhesion kinase
J. Biol. Chem.
(2000)
J.B. Aquino et al.
In vitro and in vivo differentiation of boundary cap neural crest stem cells into mature Schwann cells
Exp. Neurol.
(2006)
J. Briscoe et al.
A homeodomain protein code specifies progenitor cell identity and neuronal fate in the ventral neural tube
Cell
(2000)
J. Cai et al.
Properties of a fetal multipotent neural stem cell (NEP cell)
Dev. Biol.
(2002)
J. Dahlstrand et al.
Nestin mRNA expression correlates with the central nervous system progenitor cell state in many, but not all, regions of developing central nervous system
Brain Res. Dev. Brain Res.
(1995)
S. Fagerstrom et al.
Protein kinase C-dependent tyrosine phosphorylation of p130cas in differentiating neuroblastoma cells
J. Biol. Chem.
(1998)
V. Graham et al.
SOX2 functions to maintain neural progenitor identity
Neuron
(2003)
M.T. Harte et al.
p130Cas, a substrate associated with v-Src and v-Crk, localizes to focal adhesions and binds to focal adhesion kinase
J. Biol. Chem.
(1996)
A.W. Helms et al.
Specification of dorsal spinal cord interneurons
Curr. Opin. Neurobiol.
(2003)
D. Lohnes et al.
Developmental roles of the retinoic acid receptors
J. Steroid Biochem. Mol. Biol.
(1995)

Q. Ma et al.

Identification of neurogenin, a vertebrate neuronal determination gene

Cell

(1996)

M. Maka et al.

Identification of Sox8 as a modifier gene in a mouse model of Hirschsprung disease reveals underlying molecular defect

Dev. Biol.

(2005)

M. Nieto et al.

Neural bHLH genes control the neuronal versus glial fate decision in cortical progenitors

Neuron

(2001)

G.M. O’Neill et al.

Integrin signalling: a new Cas(t) of characters enters the stage

Trends Cell Biol.

(2000)

Y. Sun et al.

Neurogenin promotes neurogenesis and inhibits glial differentiation by independent mechanisms

Cell

(2001)

A. Wallen et al.

Orphan nuclear receptor Nurr1 is essential for Ret expression in midbrain dopamine neurons and in the brain stem

Mol. Cell. Neurosci.

(2001)

L.T. Yang et al.

c-SRC mediates neurite outgrowth through recruitment of Crk to the scaffolding protein Sin/Efs without altering the kinetics of ERK activation

J. Biol. Chem.

(2002)

A.H. Bouton et al.

Functions of the adapter protein Cas: signal convergence and the determination of cellular responses

Oncogene

(2001)

E.D. Dickman et al.

Temporally-regulated retinoic acid depletion produces specific neural crest, ocular and nervous system defects

Development

(1997)

J. Ericson et al.

Early stages of motor neuron differentiation revealed by expression of homeobox gene Islet-1

Science

(1992)

S.J. Fashena et al.

Dissection of HEF1-dependent functions in motility and transcriptional regulation

J. Cell Sci.

(2002)

Cited by (16)

Preclinical and clinical studies of the NEDD9 scaffold protein in cancer and other diseases
2015, Gene
Citation Excerpt :
A detailed discussion of NEDD9 gene and protein structure and function are beyond the scope of this article, but this topic has recently been reviewed in detail (Tikhmyanova et al., 2010; Nikonova et al., 2014a; O'Neill et al., 2000, 2007). In very brief summary, the NEDD9 promoter contains a retinoic acid response element (RARE) (Knutson and Clagett-Dame, 2015; Merrill et al., 2004a) that is specifically bound by a retinoid X receptor (RXR)/retinoic acid receptor (RAR) heterodimer (Merrill et al., 2004b), as well as xenobiotic responsive elements (Bui et al., 2009) and binding sites for transcription factors FoxC1 (Xia et al., 2013), Ngn2 (Aquino et al., 2008), HIF-1α (Kim et al., 2010), TCF (Li et al., 2011), SAFB1 (Hammerich-Hille et al., 2010), NF-Kβ, STAT5A, ER-α, GATA and others (web-links http://www.sabiosciences.com/chipqpcrsearch.php?app=TFBS and http://www.ncbi.nlm.nih.gov/gene/4739 provide useful resources) (Fig. 1A). Additionally, NEDD9 is regulated by miR-145, which binds the NEDD9 3′-untranslated region (Speranza et al., 2012; Guo et al., 2013; Lu et al., 2014).
Cancer progression requires a significant reprogramming of cellular signaling to support the essential tumor-specific processes that include hyperproliferation, invasion (for solid tumors) and survival of metastatic colonies. NEDD9 (also known as CasL and HEF1) encodes a multi-domain scaffolding protein that assembles signaling complexes regulating multiple cellular processes relevant to cancer. These include responsiveness to signals emanating from the T and B cell receptors, integrins, chemokine receptors, and receptor tyrosine kinases, as well as cytoplasmic oncogenes such as BCR-ABL and FAK- and SRC-family kinases. Downstream, NEDD9 regulation of partners including CRKL, WAVE, PI3K/AKT, ERK, E-cadherin, Aurora-A (AURKA), HDAC6, and others allow NEDD9 to influence functions as pleiotropic as migration, invasion, survival, ciliary resorption, and mitosis. In this review, we summarize a growing body of preclinical and clinical data that indicate that while NEDD9 is itself non-oncogenic, changes in expression of NEDD9 (most commonly elevation of expression) are common features of tumors, and directly impact tumor aggressiveness, metastasis, and response to at least some targeted agents inhibiting NEDD9-interacting proteins. These data strongly support the relevance of further development of NEDD9 as a biomarker for therapeutic resistance. Finally, we briefly discuss emerging evidence supporting involvement of NEDD9 in additional pathological conditions, including stroke and polycystic kidney disease.
The retinoic acid inducible Cas-family signaling protein Nedd9 regulates neural crest cell migration by modulating adhesion and actin dynamics
2009, Neuroscience
Citation Excerpt :
Isl1+/Brn3a+/Foxs1+/Sox10- post-migratory and neuronally committed progenitor cells of the DRG were negative for Nedd9 expression (percent Isl1+ cells lacking expression of Nedd9; 98.75%±1.25%, n=8; Fig. 1G, I and not shown). At E12.5, when the NCCs are committed to either glial or neuronal fates in the ventral migratory pathway, Nedd9 was no longer expressed in the neural crest/neural crest-derived cells (Aquino et al., 2008). A similar developmental sequence of Nedd9 expression in NCCs was observed in the chick (supplementary Fig. S1G, H).
Cell migration is essential for the development of numerous structures derived from embryonic neural crest cells (NCCs), however the underlying molecular mechanisms are incompletely understood. NCCs migrate long distances in the embryo and contribute to many different cell types, including peripheral neurons, glia and pigment cells. In the present work we report expression of Nedd9, a scaffolding protein within the integrin signaling pathway, in non-lineage-restricted neural crest progenitor cells. In particular, Nedd9 was found to be expressed in the dorsal neural tube at the time of neural crest delamination and in early migrating NCCs. To analyze the role of Nedd9 in neural crest development we performed loss- and gain-of-function experiments and examined the subsequent effects on delamination and migration in vitro and in vivo. Our results demonstrate that loss of Nedd9 activity in chick NCCs perturbs cell spreading and the density of focal complexes and actin filaments, properties known to depend on integrins. Moreover, a siRNA dose-dependent decrease in Nedd9 activity results in a graded reduction of NCC's migratory distance while forced overexpression increases it. Retinoic acid (RA) was found to regulate Nedd9 expression in NCCs. Our results demonstrate in vivo that Nedd9 promotes the migration of NCCs in a graded manner and suggest a role for RA in the control of Nedd9 expression levels.
dcHiC detects differential compartments across multiple Hi-C datasets
2022, Nature Communications
dcHiC: differential compartment analysis of Hi-C datasets
2022, Research Square
SOX2 promotes hypoxia-induced breast cancer cell migration by inducing NEDD9 expression and subsequent activation of Rac1/HIF-1α signaling
2019, Cellular and Molecular Biology Letters
Schwann cell precursors in health and disease
2018, GLIA

View all citing articles on Scopus

View full text

Differential expression and dynamic changes of murine NEDD9 in progenitor cells of diverse tissues

Abstract

Section snippets

Results and discussion

Materials and methods

Acknowledgements

J. Biol. Chem.

Exp. Neurol.

Cell

Dev. Biol.

Brain Res. Dev. Brain Res.

J. Biol. Chem.

Neuron

J. Biol. Chem.

Curr. Opin. Neurobiol.

J. Steroid Biochem. Mol. Biol.

Cell

Dev. Biol.

Neuron

Trends Cell Biol.

Cell

Mol. Cell. Neurosci.

J. Biol. Chem.

Functions of the adapter protein Cas: signal convergence and the determination of cellular responses

Oncogene

Temporally-regulated retinoic acid depletion produces specific neural crest, ocular and nervous system defects

Development

Early stages of motor neuron differentiation revealed by expression of homeobox gene Islet-1

Science

Dissection of HEF1-dependent functions in motility and transcriptional regulation

J. Cell Sci.