O54949 (NLK_MOUSE) Reviewed, UniProtKB/Swiss-Prot

Names and origin

Protein names

Recommended name:
Serine/threonine-protein kinase NLK
EC=2.7.11.24

Alternative name(s):
Nemo-like kinase

Gene names

Name:

Nlk

Organism

Mus musculus (Mouse) [Reference proteome]

Taxonomic identifier

10090 [NCBI]

Taxonomic lineage

cellular organisms › Eukaryota › Opisthokonta › Metazoa › Eumetazoa › Bilateria › Deuterostomia › Chordata › Craniata › Vertebrata › Gnathostomata › Teleostomi › Euteleostomi › Sarcopterygii › Tetrapoda › Amniota › Mammalia › Theria › Eutheria › Euarchontoglires › Glires › Rodentia › Sciurognathi › Muroidea › Muridae › Murinae › Mus › Mus

Protein attributes

Sequence length	527 AA.
Sequence status	Complete.
Protein existence	Evidence at protein level

General annotation (Comments)

Function	Serine/threonine-protein kinase that regulates a number of transcription factors with key roles in cell fate determination. Positive effector of the non-canonical Wnt signaling pathway, acting downstream of WNT5A, MAP3K7/TAK1 and HIPK2. Activation of this pathway causes binding to and phosphorylation of the histone methyltransferase SETDB1. The NLK-SETDB1 complex subsequently interacts with PPARG, leading to methylation of PPARG target promoters at histone H3K9 and transcriptional silencing. The resulting loss of PPARG target gene transcription inhibits adipogenesis and promotes osteoblastogenesis in mesenchymal stem cells (MSCs). Negative regulator of the canonical Wnt/beta-catenin signaling pathway. Binds to and phosphorylates TCF7L2/TCF4 and LEF1, promoting the dissociation of the TCF7L2/LEF1/beta-catenin complex from DNA, as well as the ubiquitination and subsequent proteolysis of LEF1. Together these effects inhibit the transcriptional activation of canonical Wnt/beta-catenin target genes. Negative regulator of the Notch signaling pathway. Binds to and phosphorylates NOTCH1, thereby preventing the formation of a transcriptionally active ternary complex of NOTCH1, RBPJ/RBPSUH and MAML1. Negative regulator of the MYB family of transcription factors. Phosphorylation of MYB leads to its subsequent proteolysis while phosphorylation of MYBL1 and MYBL2 inhibits their interaction with the coactivator CREBBP. Other transcription factors may also be inhibited by direct phosphorylation of CREBBP itself. Acts downstream of IL6 and MAP3K7/TAK1 to phosphorylate STAT3, which is in turn required for activation of NLK by MAP3K7/TAK1. Ref.1 Ref.4 Ref.5 Ref.6 Ref.7 Ref.8 Ref.9 Ref.10 Ref.11 Ref.12 Ref.13 Ref.15 Ref.16 Ref.17 Ref.18 Ref.19
Catalytic activity	ATP + a protein = ADP + a phosphoprotein. Ref.9 Ref.13
Cofactor	Magnesium. Ref.1 Ref.8
Enzyme regulation	Activated by the non-canonical Wnt signaling pathway, in which WNT5A leads to activation of MAP3K7/TAK1 and HIPK2, which subsequently phosphorylates and activates this protein. Activated by dimerization and subsequent intermolecular autophosphorylation on Thr-298. Other cytokines such as IL6 may also activate this regulatory circuit. Ref.1 Ref.4 Ref.6 Ref.12 Ref.19
Subunit structure	Interacts with RNF138/NARF By similarity. Forms a complex with CHD7, PPARG and SETDB1 in response to WNT5A signaling By similarity. Interacts with DAPK3/ZIPK, and this interaction may disrupt interaction with transcription factors such as TCF7L2/TCF4 By similarity. Interacts with FOXO1 and FOXO3 By similarity. Homodimer. Homodimerization is required for intermolecular autophosphorylation, kinase activation and nuclear localization. Interacts with the upstream activating kinases HIPK2 and MAP3K7/TAK1. Interaction with MAP3K7/TAK1 seems to be indirect, and may be mediated by other proteins such as STAT3, TAB1 and TAB2. Interacts with and phosphorylates a number of transcription factors including FOXO4, LEF1, MYB, MYBL1, MYBL2, NOTCH1 and TCF7L2/TCF4. May interact with components of cullin-RING-based SCF (SKP1-CUL1-F-box protein) E3 ubiquitin-protein ligase complexes. Interacts with MEF2A. Ref.7 Ref.9 Ref.10 Ref.11 Ref.12 Ref.13 Ref.15 Ref.16 Ref.17 Ref.18 Ref.19
Subcellular location	Nucleus. Cytoplasm. Note: Predominantly nuclear. A smaller fraction is cytoplasmic. Ref.1 Ref.19
Tissue specificity	Expressed at high levels in the brain, and at lower levels in heart, kidney, lung and liver. Ref.1
Domain	Contains a TQE activation loop motif in which autophosphorylation of the threonine residue (Thr-298) is sufficient for kinase activation. This mode of activation contrasts with that of classical MAP kinases, which contain a TXY activation loop motif in which phosphorylation of both the threonine and tyrosine residues is required for kinase activation.
Post-translational modification	Phosphorylated on Thr-298. Intermolecular autophosphorylation on Thr-298 activates the enzyme. Ref.1 Ref.4 Ref.16 Ref.19
Sequence similarities	Belongs to the protein kinase superfamily. CMGC Ser/Thr protein kinase family. MAP kinase subfamily. Contains 1 protein kinase domain.
Sequence caution	The sequence AAC24499.1 differs from that shown. Reason: Erroneous initiation. Translation N-terminally extended.

Ontologies

Keywords
Biological process	Transcription Transcription regulation Wnt signaling pathway
Cellular component	Cytoplasm Nucleus
Ligand	ATP-binding Magnesium Metal-binding Nucleotide-binding
Molecular function	Kinase Serine/threonine-protein kinase Transferase
PTM	Phosphoprotein
Technical term	Complete proteome Reference proteome
Gene Ontology (GO)
Biological_process	Wnt receptor signaling pathway Inferred from electronic annotation. Source: UniProtKB-KW negative regulation of Wnt receptor signaling pathway Inferred from mutant phenotype Ref.10. Source: UniProtKB peptidyl-threonine phosphorylation Inferred from direct assay Ref.13. Source: UniProtKB protein autophosphorylation Inferred from direct assay Ref.1. Source: MGI regulation of transcription, DNA-dependent Inferred from mutant phenotype Ref.8. Source: UniProtKB serine phosphorylation of STAT3 protein Inferred from direct assay Ref.9. Source: UniProtKB transcription, DNA-dependent Inferred from electronic annotation. Source: UniProtKB-KW transforming growth factor beta receptor signaling pathway Inferred from sequence or structural similarity. Source: UniProtKB
Cellular_component	cytoplasm Inferred from electronic annotation. Source: UniProtKB-SubCell nucleus Inferred from direct assay Ref.1. Source: UniProtKB
Molecular_function	ATP binding Inferred from direct assay Ref.1. Source: UniProtKB MAP kinase activity Inferred from direct assay Ref.1. Source: UniProtKB magnesium ion binding Inferred from direct assay Ref.1. Source: UniProtKB transcription factor binding Inferred from sequence or structural similarity. Source: UniProtKB ubiquitin protein ligase binding Inferred from sequence or structural similarity. Source: UniProtKB
Complete GO annotation...

Binary interactions

With	Entry	#Exp.	IntAct	Notes
Hipk2	Q9QZR5	2	EBI-366894,EBI-366905

Sequence annotation (Features)

	Feature key	Position(s)	Length	Description	Graphical view	Feature identifier
Molecule processing
	Chain	1 – 527	527	Serine/threonine-protein kinase NLK		PRO_0000186337
Regions
	Domain	138 – 427	290	Protein kinase
	Nucleotide binding	144 – 152	9	ATP By similarity
	Region	1 – 304	304	Required for interaction with TAB2
	Region	1 – 125	125	Sufficient for interaction with DAPK3 By similarity
	Region	124 – 416	293	Sufficient for interaction with DAPK3 By similarity
	Region	428 – 527	100	Required for homodimerization and kinase activation and localization to the nucleus
	Region	434 – 527	94	Required for interaction with TAB2
	Motif	298 – 300	3	TQE
	Compositional bias	22 – 25	4	Poly-Ala
	Compositional bias	27 – 34	8	Poly-His
	Compositional bias	42 – 48	7	Poly-His
	Compositional bias	71 – 83	13	Poly-Ala
	Compositional bias	106 – 111	6	Poly-Ala
	Compositional bias	115 – 119	5	Poly-Ala
Sites
	Active site	264	1	Proton acceptor By similarity
	Binding site	167	1	ATP Probable
Amino acid modifications
	Modified residue	298	1	Phosphothreonine; by autocatalysis Ref.14 Ref.19
	Modified residue	522	1	Phosphoserine By similarity
Experimental info
	Mutagenesis	167	1	K → M: Abrogates kinase activity and autophosphorylation. Retains ability to homodimerize. Abrogates ability to induce ubiquitination and degradation of LEF1 and repress canonical Wnt/beta-catenin signaling. Abrogates ability to induce MYB degradation, and reduces ability to repress MYBL1 and MYBL2. Ref.1 Ref.4 Ref.7 Ref.10 Ref.12 Ref.16 Ref.17 Ref.19
	Mutagenesis	298	1	T → D or E: Abrogates autophosphorylation. Ref.1 Ref.10 Ref.19
	Mutagenesis	298	1	T → V: Abrogates autophosphorylation. Retains ability to homodimerize. Abrogates ability to induce MYB degradation. Ref.1 Ref.10 Ref.19
	Mutagenesis	437	1	C → S: Retains kinase activity. Ref.4 Ref.19
	Mutagenesis	437	1	C → Y: Abrogates homodimerization and intermolecular autophosphorylation, with consequent loss of kinase activity. Loss of nuclear localization. Ref.4 Ref.19
	Sequence conflict	69	1	S → P in AAC24499. Ref.1

Sequences

	Sequence		Length	Mass (Da)
	O54949 [UniParc]. Last modified November 3, 2009. Version 2. Checksum: CE6D5DCCB9133989	FASTA	527	58,313
	10 20 30 40 50 60 MSLCGTRANA KMMAAYNGGT SAAAAGHHHH HHHHLPHLPP PHLHHHHHPQ HHLHPGSAAA 70 80 90 100 110 120 VHPVQQHTSS AAAAAAAAAA AAAMLNPGQQ QPYFPSPAPG QAPGPAAAAP AQVQAAAAAT 130 140 150 160 170 180 VKAHHHQHSH HPQQQLDIEP DRPIGYGAFG VVWSVTDPRD GKRVALKKMP NVFQNLVSCK 190 200 210 220 230 240 RVFRELKMLC FFKHDNVLSA LDILQPPHID YFEEIYVVTE LMQSDLHKII VSPQPLSSDH 250 260 270 280 290 300 VKVFLYQILR GLKYLHSAGI LHRDIKPGNL LVNSNCVLKI CDFGLARVEE LDESRHMTQE 310 320 330 340 350 360 VVTQYYRAPE ILMGSRHYSN AIDIWSVGCI FAELLGRRIL FQAQSPIQQL DLITDLLGTP 370 380 390 400 410 420 SLEAMRTACE GAKAHILRGP HKQPSLPVLY TLSSQATHEA VHLLCRMLVF DPSKRISAKD 430 440 450 460 470 480 ALAHPYLDEG RLRYHTCMCK CCFSTSTGRV YTSDFEPVTN PKFDDTFEKN LSSVRQVKEI 490 500 510 520 IHQFILEQQK GNRVPLCINP QSAAFKSFIS STVAQPSEMP PSPLVWE

References

	« Hide 'large scale' referencesShow 'large scale' references »
[1]	"Nlk is a murine protein kinase related to Erk/MAP kinases and localized in the nucleus." Brott B.K., Pinsky B.A., Erikson R.L. Proc. Natl. Acad. Sci. U.S.A. 95:963-968(1998) [PubMed] [Europe PMC] [Abstract] Cited for: NUCLEOTIDE SEQUENCE [MRNA], FUNCTION, ENZYME REGULATION, SUBCELLULAR LOCATION, TISSUE SPECIFICITY, AUTOPHOSPHORYLATION, MUTAGENESIS OF LYS-167 AND THR-298. Strain: BALB/c. Tissue: Brain.
[2]	"Lineage-specific biology revealed by a finished genome assembly of the mouse." Church D.M., Goodstadt L., Hillier L.W., Zody M.C., Goldstein S., She X., Bult C.J., Agarwala R., Cherry J.L., DiCuccio M., Hlavina W., Kapustin Y., Meric P., Maglott D., Birtle Z., Marques A.C., Graves T., Zhou S. , Teague B., Potamousis K., Churas C., Place M., Herschleb J., Runnheim R., Forrest D., Amos-Landgraf J., Schwartz D.C., Cheng Z., Lindblad-Toh K., Eichler E.E., Ponting C.P. PLoS Biol. 7:E1000112-E1000112(2009) [PubMed] [Europe PMC] [Abstract] Cited for: NUCLEOTIDE SEQUENCE [LARGE SCALE GENOMIC DNA]. Strain: C57BL/6J.
[3]	"The status, quality, and expansion of the NIH full-length cDNA project: the Mammalian Gene Collection (MGC)." The MGC Project Team Genome Res. 14:2121-2127(2004) [PubMed] [Europe PMC] [Abstract] Cited for: NUCLEOTIDE SEQUENCE [LARGE SCALE MRNA]. Strain: C57BL/6 and FVB/N. Tissue: Brain and Kidney.
[4]	"The TAK1-NLK-MAPK-related pathway antagonizes signalling between beta-catenin and transcription factor TCF." Ishitani T., Ninomiya-Tsuji J., Nagai S., Nishita M., Meneghini M., Barker N., Waterman M., Bowerman B., Clevers H., Shibuya H., Matsumoto K. Nature 399:798-802(1999) [PubMed] [Europe PMC] [Abstract] Cited for: FUNCTION, AUTOPHOSPHORYLATION, ENZYME REGULATION, MUTAGENESIS OF LYS-167 AND CYS-437.
[5]	"Abnormal bone marrow stroma in mice deficient for nemo-like kinase, Nlk." Kortenjann M., Nehls M., Smith A.J., Carsetti R., Schuler J., Kohler G., Boehm T. Eur. J. Immunol. 31:3580-3587(2001) [PubMed] [Europe PMC] [Abstract] Cited for: FUNCTION.
[6]	"The TAK1-NLK mitogen-activated protein kinase cascade functions in the Wnt-5a/Ca(2+) pathway to antagonize Wnt/beta-catenin signaling." Ishitani T., Kishida S., Hyodo-Miura J., Ueno N., Yasuda J., Waterman M., Shibuya H., Moon R.T., Ninomiya-Tsuji J., Matsumoto K. Mol. Cell. Biol. 23:131-139(2003) [PubMed] [Europe PMC] [Abstract] Cited for: FUNCTION, ENZYME REGULATION.
[7]	"Regulation of lymphoid enhancer factor 1/T-cell factor by mitogen-activated protein kinase-related Nemo-like kinase-dependent phosphorylation in Wnt/beta-catenin signaling." Ishitani T., Ninomiya-Tsuji J., Matsumoto K. Mol. Cell. Biol. 23:1379-1389(2003) [PubMed] [Europe PMC] [Abstract] Cited for: FUNCTION, INTERACTION WITH LEF1 AND TCF7L2, MUTAGENESIS OF LYS-167.
[8]	"Nemo-like kinase suppresses a wide range of transcription factors, including nuclear factor-kappaB." Yasuda J., Yokoo H., Yamada T., Kitabayashi I., Sekiya T., Ichikawa H. Cancer Sci. 95:52-57(2004) [PubMed] [Europe PMC] [Abstract] Cited for: FUNCTION.
[9]	"Role of the TAK1-NLK-STAT3 pathway in TGF-beta-mediated mesoderm induction." Ohkawara B., Shirakabe K., Hyodo-Miura J., Matsuo R., Ueno N., Matsumoto K., Shibuya H. Genes Dev. 18:381-386(2004) [PubMed] [Europe PMC] [Abstract] Cited for: FUNCTION, CATALYTIC ACTIVITY, INTERACTION WITH STAT3.
[10]	"Wnt-1 signal induces phosphorylation and degradation of c-Myb protein via TAK1, HIPK2, and NLK." Kanei-Ishii C., Ninomiya-Tsuji J., Tanikawa J., Nomura T., Ishitani T., Kishida S., Kokura K., Kurahashi T., Ichikawa-Iwata E., Kim Y., Matsumoto K., Ishii S. Genes Dev. 18:816-829(2004) [PubMed] [Europe PMC] [Abstract] Cited for: FUNCTION, INTERACTION WITH MYB AND HIPK2, MUTAGENESIS OF LYS-167 AND THR-298.
[11]	"Differential sensitivity of v-Myb and c-Myb to Wnt-1-induced protein degradation." Kanei-Ishii C., Nomura T., Tanikawa J., Ichikawa-Iwata E., Ishii S. J. Biol. Chem. 279:44582-44589(2004) [PubMed] [Europe PMC] [Abstract] Cited for: FUNCTION, INTERACTION WITH MYB.
[12]	"The Wnt-NLK signaling pathway inhibits A-Myb activity by inhibiting the association with coactivator CBP and methylating histone H3." Kurahashi T., Nomura T., Kanei-Ishii C., Shinkai Y., Ishii S. Mol. Biol. Cell 16:4705-4713(2005) [PubMed] [Europe PMC] [Abstract] Cited for: FUNCTION, ENZYME REGULATION, INTERACTION WITH MYBL1 AND MYBL2, MUTAGENESIS OF LYS-167.
[13]	"Nemo-like kinase-myocyte enhancer factor 2A signaling regulates anterior formation in Xenopus development." Satoh K., Ohnishi J., Sato A., Takeyama M., Iemura S., Natsume T., Shibuya H. Mol. Cell. Biol. 27:7623-7630(2007) [PubMed] [Europe PMC] [Abstract] Cited for: FUNCTION, CATALYTIC ACTIVITY, INTERACTION WITH MEF2A.
[14]	"Qualitative and quantitative analyses of protein phosphorylation in naive and stimulated mouse synaptosomal preparations." Munton R.P., Tweedie-Cullen R., Livingstone-Zatchej M., Weinandy F., Waidelich M., Longo D., Gehrig P., Potthast F., Rutishauser D., Gerrits B., Panse C., Schlapbach R., Mansuy I.M. Mol. Cell. Proteomics 6:283-293(2007) [PubMed] [Europe PMC] [Abstract] Cited for: PHOSPHORYLATION [LARGE SCALE ANALYSIS] AT THR-298, MASS SPECTROMETRY. Tissue: Brain cortex.
[15]	"Fbxw7 acts as an E3 ubiquitin ligase that targets c-Myb for nemo-like kinase (NLK)-induced degradation." Kanei-Ishii C., Nomura T., Takagi T., Watanabe N., Nakayama K.I., Ishii S. J. Biol. Chem. 283:30540-30548(2008) [PubMed] [Europe PMC] [Abstract] Cited for: FUNCTION, INTERACTION WITH E3 UBIQUITIN-PROTEIN LIGASE COMPLEXES.
[16]	"TAB2 scaffolds TAK1 and NLK in repressing canonical Wnt signaling." Li M., Wang H., Huang T., Wang J., Ding Y., Li Z., Zhang J., Li L. J. Biol. Chem. 285:13397-13404(2010) [PubMed] [Europe PMC] [Abstract] Cited for: FUNCTION, AUTOPHOSPHORYLATION, INTERACTION WITH MAP3K7 AND TAB2, MUTAGENESIS OF LYS-167.
[17]	"Nemo-like kinase suppresses Notch signalling by interfering with formation of the Notch active transcriptional complex." Ishitani T., Hirao T., Suzuki M., Isoda M., Ishitani S., Harigaya K., Kitagawa M., Matsumoto K., Itoh M. Nat. Cell Biol. 12:278-285(2010) [PubMed] [Europe PMC] [Abstract] Cited for: FUNCTION, INTERACTION WITH NOTCH1, MUTAGENESIS OF LYS-167.
[18]	"Oxidative stress-dependent regulation of Forkhead box O4 activity by nemo-like kinase." Szypowska A.A., de Ruiter H., Meijer L.A.T., Smits L.M.M., Burgering B.M.T. Antioxid. Redox Signal. 14:563-578(2011) [PubMed] [Europe PMC] [Abstract] Cited for: FUNCTION, INTERACTION WITH FOXO4.
[19]	"Homodimerization of Nemo-like kinase is essential for activation and nuclear localization." Ishitani S., Inaba K., Matsumoto K., Ishitani T. Mol. Biol. Cell 22:266-277(2011) [PubMed] [Europe PMC] [Abstract] Cited for: FUNCTION, AUTOPHOSPHORYLATION, ENZYME REGULATION, HOMODIMERIZATION, SUBCELLULAR LOCATION, PHOSPHORYLATION AT THR-298, MUTAGENESIS OF LYS-167; THR-298 AND CYS-437.
+	Additional computationally mapped references.

Cross-references

Sequence databases
EMBL GenBank DDBJ	AF036332 mRNA. Translation: AAC24499.1. Different initiation. AL591177, AL591376 Genomic DNA. Translation: CAI25549.2. AL591376, AL591177 Genomic DNA. Translation: CAI25603.2. BC057667 mRNA. Translation: AAH57667.2. BC058652 mRNA. Translation: AAH58652.2.
IPI	IPI00556914.
RefSeq	NP_032728.3. NM_008702.3.
UniGene	Mm.9001.
3D structure databases
ProteinModelPortal	O54949.
SMR	O54949. Positions 143-468.
ModBase	Search...
Protein-protein interaction databases
IntAct	O54949. 2 interactions.
PTM databases
PhosphoSite	O54949.
Proteomic databases
PaxDb	O54949.
PRIDE	O54949.
Protocols and materials databases
StructuralBiologyKnowledgebase	Search...
Genome annotation databases
Ensembl	ENSMUST00000142739; ENSMUSP00000119345; ENSMUSG00000017376.
GeneID	18099.
KEGG	mmu:18099.
UCSC	uc007kjw.1. mouse.
Organism-specific databases
CTD	51701.
MGI	MGI:1201387. Nlk.
Phylogenomic databases
eggNOG	COG0515.
GeneTree	ENSGT00550000074298.
HOGENOM	HOG000233024.
HOVERGEN	HBG014652.
InParanoid	Q5SYE6.
KO	K04468.
OMA	CKCCYTT.
Gene expression databases
ArrayExpress	O54949.
Bgee	O54949.
CleanEx	MM_NLK.
Genevestigator	O54949.
GermOnline	ENSMUSG00000017376. Mus musculus.
Family and domain databases
InterPro	IPR011009. Kinase-like_dom. IPR003527. MAP_kinase_CS. IPR000719. Prot_kinase_cat_dom. IPR017441. Protein_kinase_ATP_BS. IPR002290. Ser/Thr_dual-sp_kinase_dom. IPR008271. Ser/Thr_kinase_AS. [Graphical view]
Pfam	PF00069. Pkinase. 1 hit. [Graphical view]
SMART	SM00220. S_TKc. 1 hit. [Graphical view]
SUPFAM	SSF56112. Kinase_like. 1 hit.
PROSITE	PS01351. MAPK. 1 hit. PS00107. PROTEIN_KINASE_ATP. 1 hit. PS50011. PROTEIN_KINASE_DOM. 1 hit. PS00108. PROTEIN_KINASE_ST. 1 hit. [Graphical view]
ProtoNet	Search...
Other
ChiTaRS	NLK. mouse.
NextBio	293259.
SOURCE	Search...

Entry information

Entry name

NLK_MOUSE

Accession

Primary (citable) accession number: O54949
Secondary accession number(s): Q5SYE6, Q6PF98

Entry history

Integrated into UniProtKB/Swiss-Prot:	July 19, 2004
Last sequence update:	November 3, 2009
Last modified:	May 1, 2013
This is version 112 of the entry and version 2 of the sequence. [Complete history]

Entry status

Reviewed (UniProtKB/Swiss-Prot)

Annotation program

Chordata Protein Annotation Program