The Small Ubiquitin-like Modifiers: Established and emerging roles in diseases Mike Tatham Ron Hay...
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Transcript of The Small Ubiquitin-like Modifiers: Established and emerging roles in diseases Mike Tatham Ron Hay...
The Small Ubiquitin-like Modifiers: Established and emerging roles in diseases
Mike Tatham
Ron Hay labWellcome Trust Centre for Gene Regulation and Expression
University of Dundee
ELRIG/SLAS Drug Discovery Manchester 2012
ELRIG/SLAS Drug Discovery Manchester 2012
Phylogenetic relationship in the ubiquitin like modifier superfamily
• SUMO system is only found in Eukaryotes• Yeasts, flies and worms only express a single SUMO • Vertebrates express three paralogues SUMO-1, SUMO-
2, SUMO-3
ELRIG/SLAS Drug Discovery Manchester 2012
The SUMO conjugation system
Ubiquitination
Substrate
Ubiquitin E1 (2)Ubiquitin E2 (~20)Ubiquitin E3 (hundreds) UU
UU
Substrate
UbiquitinProtease (~100)
SS
S S
S
SUMOylation
SUMO E1 (1 – SAE1/2)SUMO E2 (1 – Ubc9)[SUMO E3 (10-20?)]
SUMOProtease (8)
SubstrateSubstrate
• SUMO system is only found in Eukaryotes• Yeasts, flies and worms only express a single SUMO • Vertebrates express three paralogues SUMO-1, SUMO-
2, SUMO-3 • Like ubiquitin SUMOs are conjugated to protein
substrates in a three step mechanism• Most SUMO conjugation occurs within a consensus
motif yKXE/D• SUMO conjugation can occur independent of E3s• SUMO-2 and SUMO-3 contain consensus motifs and
can modify themselves to form polySUMO chains• Deletion of yeast SUMO is lethal• Deletion of Ubc9 in mice is lethal
yKXE yKXD
Structural overview
• SUMO system is only found in Eukaryotes• Yeasts, flies and worms only express a single SUMO • Vertebrates express three paralogues SUMO-1, SUMO-
2, SUMO-3 • Like ubiquitin SUMOs are conjugated to protein
substrates in a three step mechanism• Most SUMO conjugation occurs within a consensus
motif yKXE/D• SUMO conjugation can occur independent of E3s• SUMO-2 and SUMO-3 contain consensus motifs and
can modify themselves to form polySUMO chains• Deletion of yeast SUMO is lethal• Deletion of Ubc9 in mice is lethal• SUMOs have low sequence homology to ubiquitin but
high 3D structural similarity
ELRIG/SLAS Drug Discovery Manchester 2012
Cellular characteristics
IB Ubiquitin
IB SUMO-2/3
IB SUMO-1
UU
UU
Substrate
U
SS
SS
Substrate
S
Substrate
S
S
Time (h)
+MG132
0 1 2 3 5 7
Total extracts
• SUMO system is only found in Eukaryotes• Yeasts, flies and worms only express a single SUMO • Vertebrates express three paralogues SUMO-1, SUMO-
2, SUMO-3 • Like ubiquitin SUMOs are conjugated to protein
substrates in a three step mechanism• Most SUMO conjugation occurs within a consensus
motif yKXE/D• SUMO conjugation can occur independent of E3s• SUMO-2 and SUMO-3 contain consensus motifs and
can modify themselves to form polySUMO chains• Deletion of yeast SUMO is lethal• Deletion of Ubc9 in mice is lethal• SUMOs have low sequence homology to ubiquitin but
high 3D structural similarity• SUMOs are predominantly nuclear proteins• SUMO-1 and SUMO-2/-3 have largely overlapping
protein targets with some distinctions
ELRIG/SLAS Drug Discovery Manchester 2012
Molecular functions of SUMO
SS
S S
S
SUMOylation
SUMO E1 (1 – SAE1/2)SUMO E2 (1 – Ubc9)[SUMO E3 (10-20?)]
SUMOProtease (8)
SubstrateSubstrate
SUMO BP
SUMO BP
polySUMO BP
Substrate
S
SSubstrate
S
Substrate
SS
S S
Subcellular localisationEnzymatic activity
Complex formationFurther modificationBlock modifications
Altered function
• SUMO system is only found in Eukaryotes• Yeasts, flies and worms only express a single SUMO • Vertebrates express three paralogues SUMO-1, SUMO-
2, SUMO-3 • Like ubiquitin SUMOs are conjugated to protein
substrates in a three step mechanism• Most SUMO conjugation occurs within a consensus
motif yKXE/D• SUMO conjugation can occur independent of E3s• SUMO-2 and SUMO-3 contain consensus motifs and
can modify themselves to form polySUMO chains• Deletion of yeast SUMO is lethal• Deletion of Ubc9 in mice is lethal• SUMOs have low sequence homology to ubiquitin but
high 3D structural similarity• SUMOs are predominantly nuclear proteins• SUMO-1 and SUMO-2/-3 have largely overlapping
protein targets with some distinctions• SUMO conjugation does not have a common effect on
proteins, but has myriad of protein-specific consequences mediated by SUMO Interaction Motifs (SIMS)
Cellular functions of SUMO
TAP-SUMO-2 cells
•Purify SUMO from cells•Identify and quantify proteins by quantitative mass spectrometry-based proteomics•Identified a total of ~900 SUMO substrates
Num
ber
of S
UM
O s
ubst
rate
s
Year
SubstrateTAG SUMO
ELRIG/SLAS Drug Discovery Manchester 2012
~10% of cellular proteins are modified by SUMO
Golebiowski et al Sci Signal. 2009;2(72):ra24. Tatham et al Sci. Signal. 2011;4(178):rs4
SUMO
ELRIG/SLAS Drug Discovery Manchester 2012
SUMO and human diseases
Immunologicaldisorders
Centromere instability, and facial anomalies syndrome
Autoimmuneregulation
Dermatomyositis
Infectiousdiseases
DNA viruses Protozoa
Extra and Intra-cellular bacteria
RNA viruses
CirculatoryDiseases
Transient global and focal cerebral ischemia
Familial dilated cardiomyopathy
Congenital heart disease
Obesity
Others
Rheumatoid arthritis
Liver damage
Cystic fibrosis
Musculardystrophy
Heart failure
Neurologicaldisorders
Multiple System Atrophy
Alzheimer's disease
Frontotemporal dementia
Spinal and bulbar muscular atrophy
Huntington's disease
Amyotrophic lateral sclerosis
Frontotemporal lobar degeneration
Spinocerebellar ataxia type 1
Neuronal intranuclear inclusion disease Dementia with Lewy Bodies
Parkinson's disease
Amyotrophic lateral sclerosis
Cancers
Megakaryoblastic leukemia
Squamous cell carcinoma
Colon cancer
Acute Promyelocytic leukaemia
Breast cancer
Ovarian cancer Multiple myeloma
MelanomaProstate cancer
Atypical myeloproliferative disease
Renal cell carcinoma
ELRIG/SLAS Drug Discovery Manchester 2012
SUMO and human diseases
Types of evidence linking SUMO with diseases1. Disease protein x is modified by SUMO which alters its function.2. SUMO conjugation is altered in disease cells3. SUMO is abnormally distributed within disease cells4. Enzymes of the SUMO modification system are abnormally expressed in disease cells5. SUMO system modulation alters the disease phenotype in model cells
Alzheimer’s disease (Amyloid beta protein (derived from from APP))(tau)
Parkinson’s disease (a-synuclein, DJ-1)Prion disease (PrP)Polyglutamine diseases Huntington’s (Huntingtin)
Kennedy’s (Androgen Rec)Dentatorubro-pallidoluysian atrophy (Atrophin-1)Spinocerebellar ataxia (ATAXIN1, 7)Tauopathy (tau)Familial amyotrophic lateral sclerosis (SOD1) Fei et al. BBRC 347 (2006)
Riley et al. JBC 280 (2005), Janer et al. Hum. Mol. Gen. 19 (2010)
Li et al. PNAS 100 (2003)
Juanes et al. JBC 284 (2009)
Mukherjee et al. JBC (2009)Steffan et al. Science (2004)
Dorval & Fraser. JBC 281 (2005), Shinbo et al. Cell. Death Diff. (2006)Dorval & Fraser. JBC 281 (2005)
Dorval & Fraser. JBC 281 (2005)
Terashima et al. Neuroreport. 13 (2002)
Immunolabelling of NII in the hippocampal subiculum of patients with NIID. (Takahashi-Fujigasaki et al. Neuropathology and Applied Neurobiology (2006), 32 , 92–100)
Immunostaining of aggregates in glioma cell models for multiple system atrophy (MSA).
(D.L. Pountney et al. Neuroscience Letters 381 (2005) 74–79)
SUMO BP
SUMO BP
ELRIG/SLAS Drug Discovery Manchester 2012
The SUMO-SIM interaction
SUMO
SIM peptide
Q. How do we take advantage of the SUMO system therapeutically?A. It depends on what you want to do!
SS
S S
SSUMO E1 (1 – SAE1/2)SUMO E2 (1 – Ubc9)[SUMO E3 (10-20?)]
SUMOProtease (8)
SubstrateSubstrateyKXE yKXD
Wimmer P, et al. J Virol. 2012 Jan;86(2):642-54.
To be or not to be specific: What can parasites tell us?
Infectiousdiseases
DNA viruses Protozoa
Extra and Intra-cellular bacteria
RNA viruses
ELRIG/SLAS Drug Discovery Manchester 2012
Q. How do we take advantage of the SUMO system therapeutically?A. It depends on how specific you want to be!
SUMO
ELRIG/SLAS Drug Discovery Manchester 2012
SUMO and human diseases
Immunologicaldisorders
Centromere instability, and facial anomalies syndrome
Autoimmuneregulation
Dermatomyositis
Infectiousdiseases
DNA viruses Protozoa
Extra and Intra-cellular bacteria
RNA viruses
CirculatoryDiseases
Transient global and focal cerebral ischemia
Familial dilated cardiomyopathy
Congenital heart disease
Obesity
Others
Rheumatoid arthritis
Liver damage
Cystic fibrosis
Musculardystrophy
Heart failure
Neurologicaldisorders
Multiple System Atrophy
Alzheimer's disease
Frontotemporal dementia
Spinal and bulbar muscular atrophy
Huntington's disease
Amyotrophic lateral sclerosis
Frontotemporal lobar degeneration
Spinocerebellar ataxia type 1
Neuronal intranuclear inclusion disease Dementia with Lewy Bodies
Parkinson's disease
Amyotrophic lateral sclerosis
Cancers
Megakaryoblastic leukemia
Squamous cell carcinoma
Colon cancer
Acute Promyelocytic leukaemia
Breast cancer
Ovarian cancer Multiple myeloma
MelanomaProstate cancer
Atypical myeloproliferative disease
Renal cell carcinoma
ELRIG/SLAS Drug Discovery Manchester 2012
Example of a successful drug therapy involving SUMO - APL
De The et al J. Cell. Biol. 2012. 198 No.1 11-21 Liu et al Curr. Op. Chem. Biol. 2012. 16 92-98
Acute Promyelocytic Leukaemia (APL)
•A rare condition driven by a chromosomal translocation resulting in the fusion of the PML and retinoic acid receptor a proteins (PML-RARa)
•Very malignant and charaterised by sudden hemorrhages and accumulation of promyelocytes in blood
•Retinoic acid and arsenic trioxide treatment induce differentiation of promyelocytes and clinical remission.
•PML-RARa and PML are known to be SUMOylated and degraded in response to arsenic
ELRIG/SLAS Drug Discovery Manchester 2012
Example of a successful drug therapy involving SUMO - APL
Ring Finger protein 4 (RNF4) aka SNURF
Tatham et al 2008. Nat. Cell. Biol. 10. 5. 538-546
ELRIG/SLAS Drug Discovery Manchester 2012
Example of a successful drug therapy involving SUMO - APL
Tatham et al 2008. Nat. Cell. Biol. 10. 5. 538-546
ELRIG/SLAS Drug Discovery Manchester 2012
A model for SUMO-dependent disease remission
PML
SUMOconjugation
SUMOdeconjugation
Ubiquitinconjugation
Ubiquitindeconjugation
U
PML
SS
S S S
PML
SS
S S SU
UUU
UUU
U
ARSENIC RNF4
Tatham et al 2008. Nat. Cell. Biol. 10. 5. 538-546
ELRIG/SLAS Drug Discovery Manchester 2012
The SUMO system as a therapeutic target - Summary
•SUMO is functionally highly pleiotropic affecting many important cellular pathways
•There is a range of evidence linking SUMO to significant human diseases.
•The precise role of SUMO in many diseases is not determined and so its potential as a therapeutic target is largely unclear
•The best approach to modulating SUMO function for individual disease therapy is unclear
•There is an academic and clinical argument for small molecule effectors of the SUMO system to help clarify these issues.
Ron HayFilip Golebiowski (Glasgow)Ellis JaffrayMarie-Claude Geoffroy (Paris)Ivan MaticAmit Garg
Jurgen CoxMatthias Mann
ELRIG/SLAS Drug Discovery Manchester 2012
Acknowledgements