Sumoylation and virus infection

1. Viral Interaction with Host cell
SUMOylation system
R. SANTHAMANIR. SANTHAMANI
I. SOPHIAI. SOPHIA
Ph.D. StudentPh.D. Student
Division of VirologyDivision of Virology
Indian Veterinary Research InstituteIndian Veterinary Research Institute
MukteswarMukteswar
Uttarakhand.Uttarakhand.

2. Contents
1. What is sumoylation?
2. Discovery
3. Sumoylation pathway
4. Importance of sumoylation
5. Immunity and sumoylation
6. Viruses and sumoylation
7. What we gain?
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3. .
Phosphorylation
Glycosylation
Ubiquitinatination
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What is sumoylation?
 SUMO – Small Ubiquitin like Modifier
 101 amino acid length
 Post translational modification
 Location: nucleus - eukaryotes
3(Saitoh et al., 1997)

4. DISCOVERY
Discovered independently
 Study of Pro-Pro and Pro-NA interactions - Yeast 2-hybrid screens
(Boddy et al., 1996; Okura et al., 1996; Shen et al., 1996)
 Biochemical analysis of nuclear transport component, RanGAP1
(Matunis et al., 1996; Mahajan et al., 1997)
 cDNA mapping of human chromosome 21
(Lapenta et al., 1997)
known by
UBL1 (Shen et al., 1996)
Sentrin (Kamitani et al., 1997)
GMP1 (Matunis et al., 1996)
hSMT3C (Lapenta etal., 1997)
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known to be proteins involved in
Sumoylation - SUMO proteins
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5. Sumoylation pathway
Between lysine residue of
substrate and C- terminal GG motif
of SUMO
Consensus motif Ψ-K-x-D/E
Between lysine residue of
substrate and C- terminal GG motif
of SUMO
Consensus motif Ψ-K-x-D/E Sumoylat
ed
protein
(Desterro et al., 1997)05/08/14 Viruses and SUMOylation 5

6. Why Sumoylation system is
needed??
 Sumolytion of vital cell
proteins pRB, PML and
RepA protein
 Targeted transport
between nucleus and
cytoplasm
 Gives stability to many
labile proteins – highly
folded structure
(Bayer et al., 1998)
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7. 05/08/14 Viruses and SUMOylation
Immunity and sumoylation…
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8.  Sumoylation of RIG 1, MDA 5 and Pellino 1
 Upregulation of IFN production
 Represses viral replication
 Sumoylation of RIG 1, MDA 5 and Pellino 1
 Upregulation of IFN production
 Represses viral replication
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Immunity and sumoylation …
(Zeng et al., 2010 & Wilson et al., 2012)
Innate immunity
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9. 05/08/14 Viruses and SUMOylation
TLR PATHWAY
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Immunity and sumoylation…
(Kim et al., 2011)

10. RLR PATHWAY
(Zeng et al., 2010)
Sumoylation of RIG 1 and MDA 5
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Immunity and sumoylation…
Type
I IFN

11.  Genetically encoded proteins - target the invading
viruses
 Expressed at a constant level
 Allows the viral infection to be halted quickly
 Genetically encoded proteins - target the invading
viruses
 Expressed at a constant level
 Allows the viral infection to be halted quickly
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Immunity and sumoylation …
Intrinsic immunity
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(Everett et al., 2009)

12.  Members of TRIM family of
proteins
 Sumoylation of TRIM family proteins play
a role in antiviral activity
 TRIM5α – retrovirus infections
TRIM5α has SIM motif to bind
with sumoylated CA of MLV
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(Nakayama et al., 2012)
Intrinsic immunity
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Immunity and sumoylation…

13. 05/08/14
Viruses and SUMOylation
(Geoffroy et al., 2011)
PML protein – Antiviral
activity
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Immunity and sumoylation…
Eg. HSV

14. 05/08/14 Viruses and SUMOylation
Viruses and Sumoylation
 Sumoylation process is exploited by viral pathogen to regulate their
activity
 Many of the DNA viruses use sumoylation for targeting their vital
proteins to the nucleus to exploit host cell nuclear machinery
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15. Virus Protein Function References
Adenovirus E1B-55K Transformation Endter et al., 2001
Adeno-associated virus Rep 78 Latency Weger et al., 2004
Epstein-Barr virus Rta (FRLF1)
EBNA3C
Lytic activation
Transcription
Chang et al., 2004
Rosendorff et al., 2004
Moloney murine
leukemia virus
CA Early stage of infection Yeuh et al., 2006
Human T cell leukemia
virus
Tax Induction of cancer Lamsoul et al., 2005
Papillomavirus E1
E2
Replication
Transcription of oncogenes
E6 , E7
Rangasamy et al., 2000
Marusic et al., 2010
Parainfluenza virus 5 P Replication Sun et al., 2011
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Sumoylation of viral proteins
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Viruses and sumoylation…

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Viruses have evolved to exploit further this intricate mechanism for
their own benefit
vProteins
 Mimic sumoylation enzymes
 Act as STUbLs
 Targets sumoylation enzymes
 Modulate sumoylation of specific host SUMO
substrates
Can viruses exploit more …? If so…Can viruses exploit more …? If so…
how???how???
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(Chang et al., 2010)
Viruses and sumoylation…

17. 05/08/14
Viral interaction with host cell sumoylation
system
vProteins that mimic
sumoylation enzymes
KSHV – KbZIP
Binds with
SUMO
Sumoylate the binding
partners - p53 & pRB
SIM
motif
SIM
motif
SUMO
ligase
(Pennella et al., 2010)
Viruses and sumoylation…
Favourable for virus?

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Viral interaction with host cell sumoylation
system
vProteins that act as STUbLs
HSV – ICP0 protein
Bind to sumoylated PML
Ubiquitination
and
proteasomal
degradation
of
PML
Act as
Ubiquitin
ligase
SIM
like
motif
SIM
like
motif
(Perry et al., 2008)
Viruses and sumoylation…

19. 05/08/14
Viral interaction with host cell sumoylation
system
vProteins that targets
sumoylation enzymes
AdenovirusAdenovirus
Sumoylation
reduced
Eg. PML
(Boggio et al., 2004)
Viruses and sumoylation…

20. 05/08/14
Viral interaction with host cell sumoylation
system
HPV L2HPV E6
Viral cycle
(Heaton et al., 2011)
Sumo ligase
Sumoylating pRB
Maintains cell
in
quiescent
Ebola
virus
VP35
Sumoylation of
IRF7
Transcriptin of
IFN reduced
(Chang et al., 2009)
Viruses and sumoylation…

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Viral interaction with host cell sumoylation
system
vProteins that modulatevProteins that modulate
sumoylation of specific hostsumoylation of specific host
SUMO substratesSUMO substrates
Adenovirus E1A,
Papilloma virus E7
pRB
Maintains cell at definite stage of cell cycle
Cell enters
S-Phase
*favourable
for virus
replication
E2F
(Ledl et al., 2005)
Viruses and sumoylation…

22. 05/08/14
Viral interaction with host cell sumoylation
system
p53 - remains in nucleus
Cell cycle arrest, apoptotic signalling
in
viral infected cells
Adenovirus E1B
Sumoylates p53 &
targets it to cytoplasm,
makes it non-
functional
Contd…
Sumo
ligase
Sumo
ligase
(Lazo et al., 2011)
Viruses and sumoylation…

23. (Wilson, 2012)05/08/14 Viruses and SUMOylation
Viruses and sumoylation :sumoylation :
In total!!In total!!

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What we gain??..
Better understanding of
 Current knowledge on virus cell interaction
 Nuclear targeting system
 Signalling systems are regulated by sumoylation
 Cellular regulatory functions
 Some aspects of pathogenesis of Cancer
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25. 05/08/14 Viruses and SUMOylation
OnlyOnly
knowledge??..knowledge??..
 SUMO gene fusion technology for expressing labile proteins like
granzyme
 If attaching SUMO to the N-terminus of poorly expressed proteins
dramatically enhances the level of expression
(Butt et al., 2005)
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 Sumoylation is an essential PTM system in eukaryotes
 Plays vital role in PTM of critical protein involved in overall cell signalling
systems including signalling pathways of intrinsic and innate immunity
 Vital for transport of proteins between cytosol and nucleus
There are established intricate connections between sumoylation, viral
infection and cancer pathogenesis
 SUMO fusion technology is a promising technique for bulk expression of
difficult to express proteins
 May be having important role in prion pathogenesis but needs to be
investigated
To conclude…To conclude…

27. 05/08/14 Viruses and SUMOylation
Future….????Future….????
 Role of Sumoylation in prion diseases?
 For bulk production of recombinant antibodies ?
Targeted delivery to nucleus by sumoylation?
 Drugs/chemicals/therapeutics with property of
controlled regulation of sumoylation for treating
cancer and viral infections?
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28. THANK
YOU
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