What Are the bona fide GSK3 Substrates?

Table 1

(a) A list of proteins reported to be substrates for GSK3. Where the phosphorylation site, priming mechanism and functional outcome of phosphorylation have been reported, this information is included. ND : not determined. (b) A list of those substrates from (a) that meet at least two out of the three criteria for confidence detailed in Section 1.3 of text, including cellular process likely to be regulated. These are the substrates discussed in more detail in the review.

(a)

Proposed substrate	Target residue(s)	Priming residue (kinase)	Effect of phosphate	Ref
Amyloid precursor protein	Thr743 (APP770)	ND	Regulates trafficking	[27–29]
	Thr668 (APP695)
APC	1501	1505 (CK1)	Regulates degradation	[30, 31]
	1503	1507 (CK1)
ATP-citrate lyase	Thr446, Ser450	Ser454 (unknown)	May regulate activity	[32, 33]
Axin	Ser322/Ser326 (putative)	Ser330	Regulates stability	[34, 35]
Axil	Not Determined		Not reported	[36]
BCL-3	Ser394,	Ser398 (ERK putative)	Regulates degradation	[37]
	Ser398 (putative)
β-catenin	Ser33, 37, Thr41	Ser45 (CK1)	Regulates degradation	[34]
δ-catenin	Thr1078 (putative)	ND	Regulates degradation	[38]
C/EBPalpha	Thr222, Thr226 (Questioned)	NONE	NONE	[39, 40]
C/EBPbeta	Ser184, Thr179	Thr188 (MAPK)	Induces DNA binding; Reduces DNA binding	[41]
	Thr189, Ser185, Ser181, Ser177	NONE		[42]
Ci-155	Ser852,	Ser856 (PKA)	Regulates Degradation	[43, 44]
	Ser884, 888	Ser892 (PKA)
CLASP	Residues between 594 and 614		Alters affinity for microtubules	[45, 46]
CLASP2	Ser533 and Ser537 (others)	Ser541 (CDK5)	Affects protein-protein interaction	[47]
CRMP2	Thr509, Thr514, Ser518	Ser522 (CDK5)	Regulates axon growth, growth cone collapse, and neuronal polarity	[48, 49]
CRMP4	Thr509, Thr514, Ser518	Ser522	Regulates axon outgrowth and chromosomal alignment	[48, 50]
CREB	Ser129	Ser133 (PKA)	Kinase activation	[51, 52]
CRY2	Ser553	Ser557	Promotes nuclear localisation and degradation	[53]
CTP: phosphocholine cytidylyltransferase	Multiple, within C-term 52 residues	?	No effect on activity	[54]
Cytidine triphosphate synthetase (CTPS)	Ser571	Ser575	Phosphorylation may reduce activity	[55]
Cyclin D1	Thr286	NONE	Nuclear export and degradation	[56]
			Questioned	[57]
Dynamin I	Thr774	Thr778 (CDK5)	Required for activity dependent bulk endocytosis	[58]
Dystrophin	ND	CKII ?	Not reported	[59]
eIF2B	Ser535	Ser539 (DYRK)	Inhibits activity	[60–62]
FAK	Ser722	Ser726	Inhibits activity	[63]
Gephyrin	Ser270	ND	Modulates GABAergic transmission	[64]
Glycogen Synthase	Ser640, 644, 648, 652	Ser656 (CKII)	Reduces activity	[65, 66]
Glucocorticoid receptor	Thr171 (Not conserved in human protein)	NONE	Inhibits GR activity towards some genes	[67]
Heat shock factor 1	Ser303	Ser307 (MAPK)	Reduces DNA binding	[68]
HIF1alpha	Ser551, Thr555, Ser589	ND	Induces proteosomal degradation	[69]
Histone H1.5	Thr10	NONE	Coincides with chromosome condensation	[70]
hnRNP D	Ser83	Ser87	Inhibits transactivation	[71]
IRS1	Ser332	Ser336	Promotes degradation	[72]
c-jun, Jun B, Jun D	Thr239	Thr243	Reduces DNA binding	[73–75]
K-casein	ND	ND	Not reported	[76]
KRP (telokin)	Ser15	Unknown site but ERK2 proposed	Not reported	[77]
LRP6	C-terminal PPPT/SP motifs (Ser1490, Thr1572, Ser1590)	NONE	Not clear	[78, 79]
MafA	Multiple sites, not identified	ND	Phosphorylation induces MafA degradation and prevents insulin gene expression	[80]
MAP1B	Ser1260, Thr1265	NONE	Regulates stability (lithium promotes degradation)	[81–83]
	Ser1388	Ser1392 (DYRK)
MAP2C	Thr1620, Thr1623	ND	Reduces microtubule binding	[84]
MARK2/PAR1	Ser212	NONE	Regulates activity	[85, 86]
Mcl1	Ser140	Thr144 (JNK)	Permits degradation of Mcl1 in response to UV stress	[87–89]
Mdm2	Ser240, Ser254	Ser244, Ser258 (CK1)	Promotes activity towards p53, reducing p53 levels	[90]
MITF	Ser298	NONE	Increases transactivation	[91]
MLK3	Ser789, Ser793	ND	Activates MLK3 Induces apoptosis in PC2 cells	[92]
MUC1/DF3	Ser40 (possible)	Ser44 (possible)	Inhibits formation of b-catenin-E-cadherin complex	[93]
c-myb	Thr572	ND	Not clear	[94, 95]
c-myc, L-myc	Thr58, Thr62 (c-myc)	Ser62 (ERK1/2)	Promotes degradation	[96–98]
Myocardin	8 serines in two blocks, Ser455—467 and Ser624—636	Yes but kinase not reported	Phosphorylation inhibits myocardin induced transcription	[99]
αNAC (nascent polypeptide associated complex)	Thr159	NONE	Induces transactivation, maybe stability	[100]
NDRG1	Thr342, Ser352, Thr362	Thr346, Thr356, Thr366 (SGK)	Not reported	[101]
neurofilament L	Ser502, 506, 603, 666 (M)	ND	Not reported	[102]
neurofilament M	Ser493 (H)			[103] (M)
neurofilament H				[104] (H)
NFAT	SRR domain	ND	Induces nuclear exclusion, inhibits DNA binding	[105–107]
	SP-2 domain	PKA or DYRK
	SP-3 domain	PKA
Ngn2	231 and 234	ND	Facilitates interaction with LIM TFs, involved in motor neuron determination	[108]
Notch 1C	ND		Stabilises protein	[109]
Nrf2	ND		Inhibits activity by nuclear exclusion	[110, 111]
OMA1	Thr339	Thr239 (MBK-2)	Induces degradation	[112]
p130Rb	Ser948	Ser952	Regulates stability
	Ser962	Ser966		[113]
	Ser982	Thr986 all CDK putative
p21 CIP1	Thr57	ND	Induces degradation	[114]
p27Kip1	Not fully established	ND	Regulates stability	[115]
p53	Ser33 (GSK3beta only)	Ser37 (DNA-PK)	Increases transcriptional activity	[116]
	Ser315, Ser376	ND	Increases cytoplasmic localisation, degradation, inhibits apoptosis	[117]
p65 RelA	Multiple, including Ser468	ND	Negatively regulates basal activity	[118, 119]
PITK	Ser1013	Ser1017 (CAMKII)	Induces nuclear localization and possibly interaction with PP1C	[120]
Polycystin-2	Ser76	Ser80	Regulates localisation, enhanced in polycystic kidney disease	[121]
PSF- Polypyrimidine tract-binding protein-associated-splicing factor	Thr687	NONE	Regulates interaction with TRAP-150, and CD45 alternative splicing in T cells	[122]
Presenilin-1	Ser397, Ser401	NONE	Reduces interaction with β-catenin	[123]
	Ser353, Ser357			[124]
Protein phosphatase1 G-subunit	Ser38, 42 (human)	Ser46 (PKA or p90RSK)	Not clear	[125]
Protein phosphatase inhibitor 2	Thr72	Ser86 (CKII)	Inhibits inhibitor, thereby activating PP1	[11, 126, 127]
PTEN	Ser362, Thr366	Ser370 (CK2)	Possibly inhibits activity	[128]
Pyruvate Dehydrogenase	ND		Inhibits activity	[129]
RCN1 (yeast calcineurn regulatory protein-calcipressin)	Ser113	Ser117	Regulates cacineurin signaling	[130]
SC35	ND	Probably	Redistributes this splicing factor	[131]
SKN-1	Ser393, (maybe Ser389 and Thr385)	Ser397	Inhibits activity	[132]
SMAD3	Thr66	ND	Regulates stability	[133]
Snail	Ser97, 101, 108, 112, 116, 120	ND	Regulates degradation and nuclear exclusion (antitumourogenic)	[134]
SREBP1c (processed fragment)	Thr426, Ser430	ND	Promotes degradation	[135]
Stathmin	Ser31	ND	Slight induction of depolymerisation of microtubules	[136]
Tau	Multiple including Ser208, Thr231, 235	Thr212 (DYRK)	Some phosphorylation sites regulate microtubule binding	[62, 137, 138]
	Ser396
	Ser404, others?			[139]
TSC2	Ser1341, Ser1337	Ser1345 (AMPK)	Activates TSC2 to inhibit mTOR	[140]
VDAC	Thr51	Thr55	Modulates interaction with HKII in mitochondrial membrane	[141]
von Hippel-Lindau (VHL)	Ser68	Ser72 (CKI)	Regulation of MT stabilization	[142]
Zcchc8	Thr492	ND	ND	[143]

(b)

Proposed substrate	Effect of phosphate	Cellular process
Amyloid precursor protein	Regulates Trafficking	Neurobiology
BCL-3	Degradation	Growth and Survival
β-catenin	Degradation	Development
C/EBPbeta	Regulates DNA binding	Endocrine control
		Growth and survival
Ci-155	Regulates degradation	Development
CLASP2	Affects protein-protein interaction	Neurobiology/cell migration
CRMP2	Regulates axon growth, growth cone collapse and neuronal polarity.	Neurobiology
CRMP4	Regulates axon outgrowth	Neurobiology
CREB	Activation	Neurobiology
		Endocrine control
Cytidine triphosphate synthetase (CTPS)	Reduces activity	Cell growth
Dynamin I	Required for activity dependent bulk endocytosis	Neurobiology
eIF2B	Inhibits activity	Cell Growth
FAK	Inhibits activity	Growth and Survival
Glycogen Synthase	Reduces activity	Endocrine control
heat shock factor 1	Reduces DNA binding	Growth and Survival
HIF1alpha	Induces proteosomal degradation	Growth and Survival
Histone H1.5	Coincides with chromosome condensation	Cell division
IRS1	Promotes degradation	Endocrine control
c-jun, Jun B, Jun D	Reduces DNA binding	Growth and survival
MAP1B	Regulates stability	Neurobiology
MAP 2C	Reduces microtubule binding	Neurobiology
MARK2/PAR1	Regulates activity	Neurobiology
Mcl1	Permits degradation of Mcl1 in response to UV stress	Growth and survival
Mdm2	Promotes activity towards p53, reduces p53 levels	Growth and survival
c-myc, L-myc	Promotes degradation	Growth and survival
Myocardin	Inhibits myocardin induced transcription	Development
NDRG1	Not reported	Ion control
NFAT	Regulates nuclear exclusion, Inhibits DNA binding	Immunology
Ngn2	Facilitates interaction with LIM TFs, for motor neuron determination	Development
p130Rb	Promotes stability.	Growth and survival
protein phosphatase 1 G-subunit	Not clear	Endocrine control
protein phosphatase inhibitor 2	Inhibits inhibitor, thereby activating PP1	Endocrine control
Polycystin-2	Regulates localisation, induced in polycystic kidney disease	Growth and survival
PTEN	Inhibits activity	Growth and survival
RCN1 (yeast calcineurn regulatory protein)	Stimulates cacineurin signaling	Growth and survival
		Neurobiology
Snail	Induces degradation and nuclear exclusion (antitumourogenic)	Growth and survival
Tau	Modulates interaction with tubulin Increased in AD	Neurobiology
VDAC	Modulates interaction with HKII in mitochondrial membrane	Growth and survival
von Hippel-Lindau (VHL)	Regulation of MT stabilization	Neurobiology