NEDD4L explained

Neural precursor cell expressed developmentally downregulated gene 4-like (NEDD4L) or NEDD4-2 is an enzyme (ubiquitin ligase) of the NEDD4 family. In human the protein is encoded by the NEDD4L gene.^{[1] [2] [3] [4]} In mouse the protein is commonly known as NEDD4-2 and the gene Nedd4-2.

NEDD4-2 has been shown to ubiquitinate and therefore down regulate the epithelial sodium channel (ENaC) in the collecting ducts of the kidneys, therefore opposing the actions of aldosterone and increasing salt excretion. In Liddle's Syndrome NEDD4 is unable to bind to the ENaC and lead to salt retention and hypertension occur.^[5]

NEDD4L belongs to the NEDD4 family of E3 HECT domain ubiquitin ligases.^{[6] [7] [8] [9]} It is the closest homologue of NEDD4, the prototypic member of the family and probably arose as a result of gene duplication. While NEDD4 orthologues are present in all eukaryotes, NEDD4L proteins are limited to vertebrates. NEDD4L proteins are known to be involved in regulating many membrane proteins via ubiquitination and endocytosis.

NEDD4L protein is expressed widely. The primary targets of NEDD4-2 include the epithelial sodium channel (ENaC), the Na+-Cl- co-transporter (NCC), and the voltage gated sodium channels (Na_vs), although additional targets are predicted from in vitro studies. NEDD4-2 gene in mice is essential for animal survival and the polymorphisms in NEDD4L are associated with human hypertension.

Protein architecture

The NEDD4-2 protein consists of an amino-terminal Ca2+-phospholipid binding domain (C2), 4 WW domains (protein-protein interaction domains) and the carboxyl-terminal HECT domain (ubiquitin ligase domain). The WW domains in the protein are responsible for binding the substrates, regulatory proteins and adaptors. These domains generally recognize PPxY (or similar) motifs in the target proteins.

Expression

Human NEDD4L gene is located on chromosome 18q12.31 with 38 exons that transcribe multiple splice variants of NEDD4L.^{[10] [11]} The protein expressed in the brain, lung, heart and the kidney contains a C2 domain. Three predominant forms of NEDD4L are isoform I containing a novel C2 domain with a start codon in exon1, isoform II with an intact conserved C2 domain consisting of an alternate start codon in exon 1 upstream of the actual start codon of the isoform 1, and isoform III lacking a C2 domain due to exon 2a–3 splicing. Isoform 1 is found to be abundant in kidney and adrenal gland whereas isoform 2 is predominantly found in the lungs.^[12] The antibodies specific to NEDD4-2 recognize two species of ~110-115 kDa in most tissues, with one being variable depending on the tissue.^[13]

Function

NEDD4L is a ubiquitin-protein ligase (E3) that accepts ubiquitin from an E2 ubiquitin-conjugating enzyme in the form of a thioester and then transfers it to specific substrates.

In vivo NEDD4-2 regulates ENaC in the lung and kidney, the renal NCC and several Navs.^{[14] [15] [16]} It has also been shown to regulate EGFR, TGFβ receptor and WNT signalling.^{[17] [18]} NEDD4L has been implicated in viral budding and viral latency processes via ubiquitination of viral proteins.^[19] In vitro data implicate NEDD4-2 in the regulation of many other proteins, including several ion channels and transporters. However most of these results have not been validated in vivo.

Regulation of NEDD4-2

NDFIP1 and NDFIP2 proteins bind NEDD4-2 and regulate its activity and/or interaction with substrates.^{[20] [21]} NEDD4-2 phosphorylation by kinases SGK1 and AKT in response to insulin and aldosterone signaling results in its interaction with 14-3-3 proteins. 14-3-3 binding to NEDD4-2 inhibits its ability to bind and ubiquitinate its substrates (such the ENaC subunits).^{[22] [23] [24] [25]} Autoubiquitination and deubiquitylation of NEDD4-2 by USP2-45 are also known to maintain NEDD4-2 protein stability.^{[26] [27]}

Clinical significance

NEDD4L is a critical regulator of renal ENaC and NCC and malfunction of this pathway has been linked to hypertension, as in Liddle's syndrome, a genetic disorder where mutations in the ENaC subunits abrogate NEDD4L binding.^{[28] [29]} In mouse, NEDD4-2 deletion leads to increased cell surface expression and activity of ENaC in the lung, resulting in premature clearance of lung fluid, airway drying, lung inflammation and perinatal lethality.^[30]

Specific deletion of NEDD4-2 in mouse renal tubules leads to increased expression of ENaC and NCC. Consistent with the critical function in ENaC and NCC regulation, NEDDL polymorphisms are linked to essential hypertension in certain human populations.^{[31] [32]} Specific deletion of NEDD4-2 in mouse neurons results in axonal branching defects.^[33] Isolated fetal cortical neurons from NEDD4-2 knockout mice show defective regulation of voltage-gated sodium currents,^[34] and in animal models of neuropathic pain NEDD4-2 expression has been found to be downregulated.^[35] Also NEDD4-2-deficiency results in hyperexcitability of DRG neurons and contributes to pathological pain^[36]

Interactions

NEDD4L has been shown to interact with SCNN1A.^[37]

Further reading

• Cook DI, Dinudom A, Komwatana P, Kumar S, Young JA . Patch-clamp studies on epithelial sodium channels in salivary duct cells . Cell Biochem. Biophys. . 36 . 2–3 . 105–13 . 2002 . 12139396 . 10.1385/CBB:36:2-3:105 . 32064659 .
• Nuber U, Schwarz S, Kaiser P, Schneider R, Scheffner M . Cloning of human ubiquitin-conjugating enzymes UbcH6 and UbcH7 (E2-F1) and characterization of their interaction with E6-AP and RSP5 . J. Biol. Chem. . 271 . 5 . 2795–800 . 1996 . 8576257 . 10.1074/jbc.271.5.2795 . free.
• Ishikawa K, Nagase T, Nakajima D, Seki N, Ohira M, Miyajima N, Tanaka A, Kotani H, Nomura N, Ohara O . Prediction of the coding sequences of unidentified human genes. VIII. 78 new cDNA clones from brain which code for large proteins in vitro . DNA Res. . 4 . 5 . 307–13 . 1997 . 9455477 . 10.1093/dnares/4.5.307 . free .
• Winberg G, Matskova L, Chen F, Plant P, Rotin D, Gish G, Ingham R, Ernberg I, Pawson T . Latent membrane protein 2A of Epstein-Barr virus binds WW domain E3 protein-ubiquitin ligases that ubiquitinate B-cell tyrosine kinases . Mol. Cell. Biol. . 20 . 22 . 8526–35 . 2000 . 11046148 . 102158 . 10.1128/MCB.20.22.8526-8535.2000 .
• Kikonyogo A, Bouamr F, Vana ML, Xiang Y, Aiyar A, Carter C, Leis J . Proteins related to the Nedd4 family of ubiquitin protein ligases interact with the L domain of Rous sarcoma virus and are required for gag budding from cells . Proc. Natl. Acad. Sci. U.S.A. . 98 . 20 . 11199–204 . 2001 . 11562473 . 58707 . 10.1073/pnas.201268998 . 2001PNAS...9811199K . free .
• Snyder PM, Olson DR, Thomas BC . Serum and glucocorticoid-regulated kinase modulates Nedd4-2-mediated inhibition of the epithelial Na+ channel . J. Biol. Chem. . 277 . 1 . 5–8 . 2002 . 11696533 . 10.1074/jbc.C100623200 . free .
• Debonneville C, Flores SY, Kamynina E, Plant PJ, Tauxe C, Thomas MA, Münster C, Chraïbi A, Pratt JH, Horisberger JD, Pearce D, Loffing J, Staub O . Phosphorylation of Nedd4-2 by Sgk1 regulates epithelial Na(+) channel cell surface expression . EMBO J. . 20 . 24 . 7052–9 . 2001 . 11742982 . 125341 . 10.1093/emboj/20.24.7052 .
• Harvey KF, Shearwin-Whyatt LM, Fotia A, Parton RG, Kumar S . N4WBP5, a potential target for ubiquitination by the Nedd4 family of proteins, is a novel Golgi-associated protein . J. Biol. Chem. . 277 . 11 . 9307–17 . 2002 . 11748237 . 10.1074/jbc.M110443200 . free .
• Chen H, Ross CA, Wang N, Huo Y, MacKinnon DF, Potash JB, Simpson SG, McMahon FJ, DePaulo Jr JR, McInnis MG . NEDD4L on human chromosome 18q21 has multiple forms of transcripts and is a homologue of the mouse Nedd4-2 gene . Eur. J. Hum. Genet. . 9 . 12 . 922–30 . 2001 . 11840194 . 10.1038/sj.ejhg.5200747 . free .
• Konstas AA, Shearwin-Whyatt LM, Fotia AB, Degger B, Riccardi D, Cook DI, Korbmacher C, Kumar S . Regulation of the epithelial sodium channel by N4WBP5A, a novel Nedd4/Nedd4-2-interacting protein . J. Biol. Chem. . 277 . 33 . 29406–16 . 2002 . 12050153 . 10.1074/jbc.M203018200 . free .
• Dunn DM, Ishigami T, Pankow J, von Niederhausern A, Alder J, Hunt SC, Leppert MF, Lalouel JM, Weiss RB . Common variant of human NEDD4L activates a cryptic splice site to form a frameshifted transcript . J. Hum. Genet. . 47 . 12 . 665–76 . 2002 . 12522688 . 10.1007/s100380200102 . free .
• Asher C, Sinha I, Garty H . Characterization of the interactions between Nedd4-2, ENaC, and sgk-1 using surface plasmon resonance . Biochim. Biophys. Acta . 1612 . 1 . 59–64 . 2003 . 12729930 . 10.1016/S0005-2736(03)00083-X .
• Itani OA, Campbell JR, Herrero J, Snyder PM, Thomas CP . Alternate promoters and variable splicing lead to hNedd4-2 isoforms with a C2 domain and varying number of WW domains . Am. J. Physiol. Renal Physiol. . 285 . 5 . F916-29 . 2003 . 12876068 . 10.1152/ajprenal.00203.2003 . 1429872 .
• Boehmer C, Henke G, Schniepp R, Palmada M, Rothstein JD, Bröer S, Lang F . Regulation of the glutamate transporter EAAT1 by the ubiquitin ligase Nedd4-2 and the serum and glucocorticoid-inducible kinase isoforms SGK1/3 and protein kinase B . J. Neurochem. . 86 . 5 . 1181–8 . 2003 . 12911626 . 10.1046/j.1471-4159.2003.01937.x . 20290123 . free .
• Malbert-Colas L, Nicolas G, Galand C, Lecomte MC, Dhermy D . Identification of new partners of the epithelial sodium channel alpha subunit . Comptes Rendus Biologies . 326 . 7 . 615–24 . 2003 . 14556380 . 10.1016/s1631-0691(03)00154-9 .
• Qi H, Grenier J, Fournier A, Labrie C . Androgens differentially regulate the expression of NEDD4L transcripts in LNCaP human prostate cancer cells . Mol. Cell. Endocrinol. . 210 . 1–2 . 51–62 . 2003 . 14615060 . 10.1016/j.mce.2003.08.009 . 33109822 .

Notes and References

1. Erdeniz N, Rothstein R . Rsp5, a ubiquitin-protein ligase, is involved in degradation of the single-stranded-DNA binding protein rfa1 in Saccharomyces cerevisiae . Mol. Cell. Biol. . 20 . 1 . 224–32 . Jan 2000 . 10594025 . 85078 . 10.1128/MCB.20.1.224-232.2000 .
2. Harvey KF, Dinudom A, Cook DI, Kumar S . The Nedd4-like protein KIAA0439 is a potential regulator of the epithelial sodium channel . J. Biol. Chem. . 276 . 11 . 8597–601 . May 2001 . 11244092 . 10.1074/jbc.C000906200 . free .
3. Raikwar NS, Thomas CP . Nedd4-2 isoforms ubiquitinate individual epithelial sodium channel subunits and reduce surface expression and function of the epithelial sodium channel . Am. J. Physiol. Renal Physiol. . 294 . 5 . F1157-65 . May 2008 . 18322022 . 2424110 . 10.1152/ajprenal.00339.2007 .
4. Web site: Entrez Gene: NEDD4L Neural precursor cell expressed, developmentally down-regulated 4-like.
5. Rotin D . Role of the UPS in Liddle syndrome . BMC Biochem. . 9 . S5 . 2008 . Suppl 1 . 19007435 . 2582799 . 10.1186/1471-2091-9-S1-S5 . free .
6. Harvey KF, Kumar S . Nedd4-like proteins: an emerging family of ubiquitin-protein ligases implicated in diverse cellular functions . Trends Cell Biol. . 9 . 5 . 166–9 . May 1999 . 10322449 . 10.1016/s0962-8924(99)01541-x .
7. Scheffner M, Kumar S . Mammalian HECT ubiquitin-protein ligases: biological and pathophysiological aspects . Biochim. Biophys. Acta . 1843 . 1 . 61–74 . Jan 2014 . 23545411 . 10.1016/j.bbamcr.2013.03.024 . free .
8. Yang B, Kumar S . Nedd4 and Nedd4-2: closely related ubiquitin-protein ligases with distinct physiological functions . Cell Death Differ. . 17 . 1 . 68–77 . Jan 2010 . 19557014 . 10.1038/cdd.2009.84 . 2818775.
9. Rotin D, Kumar S . Physiological functions of the HECT family of ubiquitin ligases . Nat. Rev. Mol. Cell Biol. . 10 . 6 . 398–409 . Jun 2009 . 19436320 . 10.1038/nrm2690 . 1894155 .
10. Araki N, Umemura M, Miyagi Y, Yabana M, Miki Y, Tamura K, Uchino K, Aoki R, Goshima Y, Umemura S, Ishigami T . Expression, transcription, and possible antagonistic interaction of the human Nedd4L gene variant: implications for essential hypertension . Hypertension . 51 . 3 . 773–7 . Mar 2008 . 18268134 . 10.1161/HYPERTENSIONAHA.107.102061 . free .
11. Itani OA, Stokes JB, Thomas CP . Nedd4-2 isoforms differentially associate with ENaC and regulate its activity . Am. J. Physiol. Renal Physiol. . 289 . 2 . F334-46 . Aug 2005 . 15814530 . 10.1152/ajprenal.00394.2004 . 17862503 .
12. Itani OA, Campbell JR, Herrero J, Snyder PM, Thomas CP . Alternate promoters and variable splicing lead to hNedd4-2 isoforms with a C2 domain and varying number of WW domains . Am. J. Physiol. Renal Physiol. . 285 . 5 . F916-29 . Nov 2003 . 12876068 . 10.1152/ajprenal.00203.2003 . 1429872 .
13. Ronzaud C, Loffing-Cueni D, Hausel P, Debonneville A, Malsure SR, Fowler-Jaeger N, Boase NA, Perrier R, Maillard M, Yang B, Stokes JB, Koesters R, Kumar S, Hummler E, Loffing J, Staub O . Renal tubular NEDD4-2 deficiency causes NCC-mediated salt-dependent hypertension . J. Clin. Invest. . 123 . 2 . 657–65 . Feb 1, 2013 . 23348737 . 10.1172/JCI61110 . 3561795.
14. Fotia AB, Ekberg J, Adams DJ, Cook DI, Poronnik P, Kumar S . Regulation of neuronal voltage-gated sodium channels by the ubiquitin-protein ligases Nedd4 and Nedd4-2 . J. Biol. Chem. . 279 . 28 . 28930–5 . Jul 9, 2004 . 15123669 . 10.1074/jbc.M402820200 . free .
15. Kamynina E, Debonneville C, Bens M, Vandewalle A, Staub O . A novel mouse Nedd4 protein suppresses the activity of the epithelial Na+ channel . FASEB J. . 15 . 1 . 204–214 . Jan 2001 . 11149908 . 10.1096/fj.00-0191com . free . 19008349 .
16. Ekberg J, Schuetz F, Boase NA, Conroy SJ, Manning J, Kumar S, Poronnik P, Adams DJ . Regulation of the voltage-gated K(+) channels KCNQ2/3 and KCNQ3/5 by ubiquitination. Novel role for Nedd4-2 . J. Biol. Chem. . 282 . 16 . 12135–42 . Apr 20, 2007 . 17322297 . 10.1074/jbc.M609385200 . free .
17. Gao S, Alarcón C, Sapkota G, Rahman S, Chen PY, Goerner N, Macias MJ, Erdjument-Bromage H, Tempst P, Massagué J . Ubiquitin ligase Nedd4L targets activated Smad2/3 to limit TGF-beta signaling . Mol. Cell . 36 . 3 . 457–68 . Nov 13, 2009 . 19917253 . 10.1016/j.molcel.2009.09.043 . 2796330.
18. Zhang Y, Ding Y, Chen YG, Tao Q . NEDD4L regulates convergent extension movements in Xenopus embryos via Disheveled-mediated non-canonical Wnt signaling . Dev. Biol. . 392 . 1 . 15–25 . Aug 1, 2014 . 24833518 . 10.1016/j.ydbio.2014.05.003 . free .
19. Chung HY, Morita E, von Schwedler U, Müller B, Kräusslich HG, Sundquist WI . NEDD4L overexpression rescues the release and infectivity of human immunodeficiency virus type 1 constructs lacking PTAP and YPXL late domains . J. Virol. . 82 . 10 . 4884–97 . May 2008 . 18321968 . 10.1128/JVI.02667-07 . 2346761.
20. Mund T, Pelham HR . Control of the activity of WW-HECT domain E3 ubiquitin ligases by NDFIP proteins . EMBO Rep. . 10 . 5 . 501–7 . May 2009 . 19343052 . 10.1038/embor.2009.30 . 2680872.
21. Shearwin-Whyatt L, Dalton HE, Foot N, Kumar S . Regulation of functional diversity within the Nedd4 family by accessory and adaptor proteins . BioEssays . 28 . 6 . 617–28 . Jun 2006 . 16700065 . 10.1002/bies.20422 .
22. Snyder PM, Olson DR, Kabra R, Zhou R, Steines JC . cAMP and serum and glucocorticoid-inducible kinase (SGK) regulate the epithelial Na(+) channel through convergent phosphorylation of Nedd4-2 . J. Biol. Chem. . 279 . 44 . 45753–8 . Oct 29, 2004 . 15328345 . 10.1074/jbc.M407858200 . free .
23. Bhalla V, Daidié D, Li H, Pao AC, LaGrange LP, Wang J, Vandewalle A, Stockand JD, Staub O, Pearce D . Serum- and glucocorticoid-regulated kinase 1 regulates ubiquitin ligase neural precursor cell-expressed, developmentally down-regulated protein 4-2 by inducing interaction with 14-3-3 . Mol. Endocrinol. . 19 . 12 . 3073–84 . Dec 2005 . 16099816 . 10.1210/me.2005-0193 . 11498723 . free .
24. Lee IH, Dinudom A, Sanchez-Perez A, Kumar S, Cook DI . Akt mediates the effect of insulin on epithelial sodium channels by inhibiting Nedd4-2 . J. Biol. Chem. . 282 . 41 . 29866–73 . Oct 12, 2007 . 17715136 . 10.1074/jbc.M701923200 . free .
25. Nagaki K, Yamamura H, Shimada S, Saito T, Hisanaga S, Taoka M, Isobe T, Ichimura T . 14-3-3 Mediates phosphorylation-dependent inhibition of the interaction between the ubiquitin E3 ligase Nedd4-2 and epithelial Na+ channels . Biochemistry . 45 . 21 . 6733–40 . May 30, 2006 . 16716084 . 10.1021/bi052640q .
26. Bruce MC, Kanelis V, Fouladkou F, Debonneville A, Staub O, Rotin D . Regulation of Nedd4-2 self-ubiquitination and stability by a PY motif located within its HECT-domain . Biochem. J. . 415 . 1 . 155–63 . Oct 1, 2008 . 18498246 . 10.1042/BJ20071708 .
27. Oberfeld B, Ruffieux-Daidié D, Vitagliano JJ, Pos KM, Verrey F, Staub O . Ubiquitin-specific protease 2-45 (Usp2-45) binds to epithelial Na+ channel (ENaC)-ubiquitylating enzyme Nedd4-2 . Am. J. Physiol. Renal Physiol. . 301 . 1 . F189-96 . Jul 2011 . 21478478 . 10.1152/ajprenal.00487.2010 .
28. Boase NA, Rychkov GY, Townley SL, Dinudom A, Candi E, Voss AK, Tsoutsman T, Semsarian C, Melino G, Koentgen F, Cook DI, Kumar S . Respiratory distress and perinatal lethality in Nedd4-2-deficient mice . Nat Commun . 2 . 4 . 287 . 2011 . 21505443 . 10.1038/ncomms1284 . 3104547. 2011NatCo...2..287B .
29. Rotin D, Staub O . Nedd4-2 and the regulation of epithelial sodium transport . Front Physiol . 3 . 212 . 2012 . 22737130 . 10.3389/fphys.2012.00212 . 3380336. free .
30. Kimura T, Kawabe H, Jiang C, Zhang W, Xiang YY, Lu C, Salter MW, Brose N, Lu WY, Rotin D . Deletion of the ubiquitin ligase Nedd4L in lung epithelia causes cystic fibrosis-like disease . Proc. Natl. Acad. Sci. U.S.A. . 108 . 8 . 3216–21 . Feb 22, 2011 . 21300902 . 10.1073/pnas.1010334108 . 3044364. 2011PNAS..108.3216K . free .
31. Graham L, Padmanabhan S . NEDD4L in essential hypertension . J. Hypertens. . 32 . 2 . 230–2 . Feb 2014 . 24430119 . 10.1097/HJH.0000000000000105 . free .
32. Russo CJ, Melista E, Cui J, DeStefano AL, Bakris GL, Manolis AJ, Gavras H, Baldwin CT . Association of NEDD4L ubiquitin ligase with essential hypertension . Hypertension . 46 . 3 . 488–91 . Sep 2005 . 16103266 . 10.1161/01.HYP.0000178594.63193.c0 . free .
33. Hsia HE, Kumar R, Luca R, Takeda M, Courchet J, Nakashima J, Wu S, Goebbels S, An W, Eickholt BJ, Polleux F, Rotin D, Wu H, Rossner MJ, Bagni C, Rhee JS, Brose N, Kawabe H . Ubiquitin E3 ligase Nedd4-1 acts as a downstream target of PI3K/PTEN-mTORC1 signaling to promote neurite growth . Proc. Natl. Acad. Sci. U.S.A. . 111 . 36 . 13205–10 . Sep 9, 2014 . 25157163 . 10.1073/pnas.1400737111 . 4246980. 2014PNAS..11113205H . free .
34. Ekberg JA, Boase NA, Rychkov G, Manning J, Poronnik P, Kumar S . Nedd4-2 (NEDD4L) controls intracellular Na(+)-mediated activity of voltage-gated sodium channels in primary cortical neurons . Biochem. J. . 457 . 1 . 27–31 . Jan 1, 2014 . 24152020 . 10.1042/BJ20131275 .
35. Laedermann CJ, Cachemaille M, Kirschmann G, Pertin M, Gosselin RD, Chang I, Albesa M, Towne C, Schneider BL, Kellenberger S, Abriel H, Decosterd I . Dysregulation of voltage-gated sodium channels by ubiquitin ligase NEDD4-2 in neuropathic pain . J. Clin. Invest. . 123 . 7 . 3002–13 . Jul 1, 2013 . 23778145 . 10.1172/JCI68996 . 3696561.
36. Cachemaille M, Laedermann CJ, Pertin M, Abriel H, Gosselin RD, Decosterd I . Neuronal expression of the ubiquitin ligase Nedd4-2 in rat dorsal root ganglia: modulation in the spared nerve injury model of neuropathic pain . Neuroscience . 227 . 370–80 . Dec 27, 2012 . 23022218 . 10.1016/j.neuroscience.2012.09.044 . 26108834 .
37. Malbert-Colas L, Nicolas G, Galand C, Lecomte MC, Dhermy D . Identification of new partners of the epithelial sodium channel alpha subunit . Comptes Rendus Biologies . 326 . 7 . 615–24 . Jul 2003 . 14556380 . 10.1016/s1631-0691(03)00154-9 .