ReviewNEDD4: The founding member of a family of ubiquitin-protein ligases
Introduction
Ubiquitination is a post-translational protein modification that is critical for a number of cellular processes. Ubiquitination involves the covalent attachment of the 8 kDa protein ubiquitin to one or more lysine residues in the substrate protein to signal proteins for degradation, altered localisation, trafficking or function. Substrate proteins can be mono-ubiquitinated, multi-monoubiquitinated or poly-ubiquitinated, with the type of ubiquitination determining the fate of the protein. Ubiquitin itself has seven lysine residues, allowing for different ubiquitin linkage types; for example the well-studied K48-linkage typically targets proteins for proteasomal degradation (Hershko and Ciechanover, 1998) whereas K63 linkages are associated with protein trafficking and lysosomal degradation (Hicke and Dunn, 2003).
Ubiquitin is covalently attached to a protein substrate via an energy dependent three step process, involving an E1 ubiquitin activating enzyme, an E2 ubiquitin conjugating enzyme and an E3 ubiquitin protein ligase. The E3 ubiquitin ligase largely determines the substrate specificity of the system and in mammals there are several hundred ubiquitin protein ligases (Hershko and Ciechanover, 1998). These can be grouped into two main classes; the RING (Really Interesting New Gene) E3s which mediate the direct transfer of ubiquitin to the substrate (Deshaies and Joazeiro, 2009), and the HECT (Homologous to E6-AP C-Terminus) E3s which are involved in the transfer of activated ubiquitin from the E2 to the substrate by forming an intermediate complex with the C-terminus of the E3 (Rotin and Kumar, 2009). This review will focus on the HECT type ubiquitin ligase NEDD4, one of the first HECT E3 ligases discovered, and the founding member of the NEDD4 family of HECT ubiquitin ligases.
Section snippets
History of NEDD4 discovery
The NEDD4 gene was cloned in 1992 as one of a number of murine Nedd (Neural precursor cell expressed developmentally down-regulated) genes differentially expressed in the central nervous system (Kumar et al., 1992). At the time of its cloning, the predicted protein had only one known domain — an N-terminal calcium/lipid-binding domain (C2 domain). The presence of three partial repeats of approximately 40 amino acids containing two conserved tryptophan residues in the middle part of the protein
NEDD4 orthologues and structure
As mentioned above, NEDD4 is a highly evolutionarily conserved protein from yeast to man, and was initially cloned as a highly expressed gene in the early embryonic brain (Kumar et al., 1992, Kumar et al., 1997). There are 94 orthologues of NEDD4 in the NCBI database, all sharing the same modular structure consisting of an N-terminal C2 domain, 3–4 WW domains and a C-terminal catalytic HECT domain for ubiquitin protein ligation (Harvey and Kumar, 1999) (Fig. 1A). The C2 domain is a
NEDD4 binding partners and targets
A number of in vitro binding studies and proteomic approaches have been used to identify potential NEDD4 substrates (see below for a summary and Table 1 for a list of interacting proteins).
Physiological functions of NEDD4
NEDD4 is widely expressed in mammalian tissues. To investigate the physiological functions of NEDD4, a number of studies have focussed on NEDD4−/− mice that lack NEDD4 expression. The first described NEDD4−/− mice were neonatal lethal, with delayed embryonic development and reduced growth and body weight due (Cao et al., 2008). Consistent with this, NEDD4−/− murine embryonic fibroblasts (MEFs) display reduced mitogenic activity (Cao et al., 2008). NEDD4−/− embryos have reduced skeletal muscle
NEDD4 auto-inhibition
NEDD4 activity is in part regulated by auto-inhibition. In the absence of calcium, an auto-inhibitory conformation of NEDD4 is formed by the C2 domain of NEDD4 binding to the HECT domain via intramolecular interactions, thereby inhibiting the enzymatic activity of NEDD4 (Wang et al., 2010). In the presence of calcium, the binding of the C2 domain to the HECT domain is disrupted and the C2 domain recruits NEDD4 to the lipid membrane promoting its ubiquitin ligase activity (Wang et al., 2010).
Conclusions
NEDD4 is the founding member of the NEDD4 family of ubiquitin protein ligases that function in the ubiquitin proteasome system. NEDD4 has many important cellular functions, as indicated by the high degree of evolutionary conservation observed across many species. NEDD4 has roles in regulating viral budding, IGF-1 signalling, in T-cell function and in PTEN signalling, although this is yet to be fully understood as there are currently a number of conflicting models. The role of NEDD4 in cancer
Acknowledgements
The ubiquitin work in our laboratory is supported by the National Health and Medical Research Council (NHMRC) Project Grants 1020755 & 1059393, and a NHMRC Senior Principal Research Fellowship 1002863 to SK.
References (121)
- et al.
The cyclin-dependent kinase activator, Spy1A, is targeted for degradation by the ubiquitin ligase NEDD4
J. Biol. Chem.
(2009) - et al.
Damage-induced ubiquitylation of human RNA polymerase II by the ubiquitin ligase Nedd4, but not Cockayne syndrome proteins or BRCA1
Mol. Cell
(2007) - et al.
The WW domain: a signalling site in dystrophin?
Trends Biochem. Sci.
(1994) - et al.
Subtle gait abnormalities in Nedd4 heterozygous mice
Behav. Brain Res.
(2014) - et al.
Disruption of E3 ligase NEDD4 in peripheral neurons interrupts axon outgrowth: linkage to PTEN
Mol. Cell. Neurosci.
(2012) - et al.
Brucella infection inhibits macrophages apoptosis via Nedd4-dependent degradation of calpain2
Vet. Microbiol.
(2014) - et al.
Nedd4 and Nedd4-2: ubiquitin ligases at work in the neuron
Int. J. Biochem. Cell Biol.
(2013) - et al.
E3 ligase Nedd4 promotes axon branching by downregulating PTEN
Neuron
(2010) - et al.
NEDD4 is overexpressed in colorectal cancer and promotes colonic cell growth independently of the PI3K/PTEN/AKT pathway
Cell. Signal.
(2013) - et al.
Ubiquitin-dependent regulation of phospho-AKT dynamics by the ubiquitin E3 ligase, NEDD4-1, in the insulin-like growth factor-1 response
J. Biol. Chem.
(2013)
Regulation of the divalent metal ion transporter DMT1 and iron homeostasis by a ubiquitin-dependent mechanism involving Ndfips and WWP2
Blood
Ndfip1-deficient mice have impaired DMT1 regulation and iron homeostasis
Blood
Regulation of neuronal voltage-gated sodium channels by the ubiquitin-protein ligases Nedd4 and Nedd4-2
J. Biol. Chem.
The ubiquitin-protein ligases Nedd4 and Nedd4-2 show similar ubiquitin-conjugating enzyme specificities
Int. J. Biochem. Cell Biol.
The ubiquitin ligase Nedd4-1 is required for heart development and is a suppressor of thrombospondin-1
J. Biol. Chem.
Eps15 interacts with ubiquitinated Cx43 and mediates its internalization
Exp. Cell Res.
E3 ubiquitin ligase Cbl-b regulates Pten via Nedd4 in T cells independently of its ubiquitin ligase activity
Cell Rep.
Nedd4-like proteins: an emerging family of ubiquitin-protein ligases implicated in diverse cellular functions
Trends Cell Biol.
The Nedd4-like protein KIAA0439 is a potential regulator of the epithelial sodium channel
J. Biol. Chem.
The Epstein–Barr virus latent membrane protein 2A PY motif recruits WW domain-containing ubiquitin-protein ligases
Virology
Regulation of Rap2A by the ubiquitin ligase Nedd4-1 controls neurite development
Neuron
Identification of a set of genes with developmentally down-regulated expression in the mouse brain
Biochem. Biophys. Res. Commun.
cDNA cloning, expression analysis, and mapping of the mouse Nedd4 gene
Genomics
Functional interaction of phosphatase and tensin homologue (PTEN) with the E3 ligase NEDD4-1 during neuronal response to zinc
J. Biol. Chem.
Abnormal development of the neuromuscular junction in Nedd4-deficient mice
Dev. Biol.
WW domain HECT E3s target Cbl RING finger E3s for proteasomal degradation
J. Biol. Chem.
ISG15 inhibits Nedd4 ubiquitin E3 activity and enhances the innate antiviral response
J. Biol. Chem.
mGrb10 interacts with Nedd4
J. Biol. Chem.
Grb10 prevents Nedd4-mediated vascular endothelial growth factor receptor-2 degradation
J. Biol. Chem.
Nedd4 family-interacting protein 1 (Ndfip1) is required for the exosomal secretion of Nedd4 family proteins
J. Biol. Chem.
gamma2-Adaptin, a ubiquitin-interacting adaptor, is a substrate to coupled ubiquitination by the ubiquitin ligase Nedd4 and functions in the endosomal pathway
J. Biol. Chem.
Neuronal precursor cell-expressed developmentally down-regulated 4-1 (NEDD4-1) controls the sorting of newly synthesized Ca(V)1.2 calcium channels
J. Biol. Chem.
Drosophila Nedd4 regulates endocytosis of notch and suppresses its ligand-independent activation
Curr. Biol.
Mammalian HECT ubiquitin-protein ligases: biological and pathophysiological aspects
Biochim. Biophys. Acta
The HPV-16 E6 and E6-AP complex functions as a ubiquitin-protein ligase in the ubiquitination of p53
Cell
Abnormal splicing of NEDD4 in myotonic dystrophy type 2: possible link to statin adverse reactions
Am. J. Pathol.
Tsg101 and Alix interact with murine leukemia virus Gag and cooperate with Nedd4 ubiquitin ligases during budding
J. Biol. Chem.
Human ubiquitin-protein ligase Nedd4: expression, subcellular localization and selective interaction with ubiquitin-conjugating enzymes
Genes Cells
The anti-proliferative response of indole-3-carbinol in human melanoma cells is triggered by an interaction with NEDD4–1 and disruption of wild-type PTEN degradation
Mol. Cancer Res.
Respiratory distress and perinatal lethality in Nedd4-2-deficient mice
Nat. Commun.
Nedd4 controls animal growth by regulating IGF-1 signaling
Sci. Signal.
The Nedd4-like family of E3 ubiquitin ligases and cancer
Cancer Metastasis Rev.
FoxM1B regulates NEDD4-1 expression, leading to cellular transformation and full malignant phenotype in immortalized human astrocytes
Cancer Res.
Drosophila Ndfip is a novel regulator of Notch signaling
Cell Death Differ.
RING domain E3 ubiquitin ligases
Annu. Rev. Biochem.
Nedd4 mediates control of an epithelial Na + channel in salivary duct cells by cytosolic Na +
Proc. Natl. Acad. Sci. U. S. A.
The C2 domain of the Rsp5 ubiquitin ligase binds membrane phosphoinositides and directs ubiquitination of endosomal cargo
J. Cell Biol.
Dysregulation of T lymphocyte function in itchy mice: a role for Itch in TH2 differentiation
Nat. Immunol.
NEDD4 ubiquitinates TRAF3 to promote CD40-mediated AKT activation
Nat. Commun.
Patched dependence receptor triggers apoptosis through ubiquitination of caspase-9
Proc. Natl. Acad. Sci. U. S. A.
Cited by (114)
An insulin-regulated arrestin domain protein controls hepatic glucagon action
2023, Journal of Biological ChemistryThe role of the NDRG1 in the pathogenesis and treatment of breast cancer
2023, Biochimica et Biophysica Acta - Reviews on CancerCelastrol targeting Nedd4 reduces Nrf2-mediated oxidative stress in astrocytes after ischemic stroke
2023, Journal of Pharmaceutical Analysis