Ubiquitin is directly linked via an ester to protein-conjugated mono-ADP-ribose
The EMBO Journal,
Год журнала:
2025,
Номер
unknown
Опубликована: Фев. 25, 2025
Abstract
The
prevailing
view
on
post-translational
modifications
(PTMs)
is
that
a
single
amino
acid
modified
with
PTM
at
any
given
time.
However,
recent
work
has
demonstrated
crosstalk
between
different
PTMs,
some
occurring
the
same
residue.
Such
interplay
seen
ADP-ribosylation
and
ubiquitylation.
For
example,
DELTEX
E3
ligases
were
reported
to
ubiquitylate
hydroxyl
group
free
NAD
+
ADP-ribose
in
vitro,
generating
noncanonical
ubiquitin
ester-linked
species.
In
this
report,
we
show,
for
first
time,
dual
occurs
cells
mono-ADP-ribosylated
(MARylated)
PARP10
Glu/Asp
sites
form
MAR
ester.
We
call
process
mono-ADP-ribosyl
ubiquitylation
or
MARUbylation.
Using
chemical
enzymatic
treatments,
including
newly
characterized
bacterial
deubiquitinase
esterase-specific
activity,
discovered
multiple
PARPs
are
MARUbylated
extended
K11-linked
polyubiquitin
chains
when
exogenously
expressed.
Finally,
show
response
type
I
interferon
stimulation,
MARUbylation
can
occur
endogenously
PARP
targets.
Thus,
represents
new
broadens
our
understanding
of
function
PARP-mediated
cells.
Язык: Английский
PARPs and ADP-ribosylation-mediated biomolecular condensates: determinants, dynamics, and disease implications
Trends in Biochemical Sciences,
Год журнала:
2025,
Номер
unknown
Опубликована: Фев. 1, 2025
Язык: Английский
Mutation of a highly conserved isoleucine residue in loop 2 of several 𝛽-coronavirus macrodomains indicates that enhanced ADP-ribose binding is detrimental to infection
bioRxiv (Cold Spring Harbor Laboratory),
Год журнала:
2024,
Номер
unknown
Опубликована: Янв. 4, 2024
All
coronaviruses
(CoVs)
encode
for
a
conserved
macrodomain
(Mac1)
located
in
nonstructural
protein
3
(nsp3).
Mac1
is
an
ADP-ribosylhydrolase
that
binds
and
hydrolyzes
mono-ADP-ribose
from
target
proteins.
Previous
work
has
shown
important
virus
replication
pathogenesis.
Within
Mac1,
there
are
several
regions
highly
across
CoVs,
including
the
GIF
(glycine-isoleucine-phenylalanine)
motif.
To
determine
how
biochemical
activities
of
these
residues
impact
CoV
replication,
isoleucine
phenylalanine
were
mutated
to
alanine
(I-A/F-A)
both
recombinant
proteins
murine
hepatitis
(MHV),
Middle
East
respiratory
syndrome
coronavirus
(MERS-CoV),
severe
acute
2
(SARS-CoV-2).
The
F-A
mutant
had
ADP-ribose
binding
and/or
hydrolysis
defects
led
attenuated
pathogenesis
cell
culture
mice.
In
contrast,
I-A
mutations
normal
enzyme
activity
enhanced
binding.
Despite
increased
binding,
MERS-CoV
SARS-CoV-2
mice,
indicating
this
residue
acts
as
gate
controls
efficient
replication.
These
results
highlight
function
provide
unique
insight
into
macrodomains
control
promote
viral
Язык: Английский
Exacerbated salmonellosis in poly(ADP-ribose) polymerase 14 deficient mice
bioRxiv (Cold Spring Harbor Laboratory),
Год журнала:
2025,
Номер
unknown
Опубликована: Янв. 2, 2025
ABSTRACT
Salmonella
enterica
subspecies
serovar
Typhimurium
(
S.
Typhimurium)
is
an
enteropathogen
annually
causing
millions
of
acute
infections
ranging
from
gastroenteritis
to
life-threatening
invasive
systemic
disease.
Strong
mucosal
inflammation
characteristic
for
the
infection
driven
by
innate
immune
receptor
activation
but
also
directly
invading
bacterium.
It
remains
unknown
how
and
at
which
point
turns
anti-bacterial
tissue
homeostasis
restored.
Here,
we
investigated
expression
function
poly(ADP-ribose)polymerase
(Parp14),
a
known
cytoplasmic
nuclear
regulatory
protein
cells,
in
mouse
streptomycin-pretreatment
model
infection.
In
infected
mice,
Parp14
expressing
some
were
macrophages,
detected
throughout
gastrointestinal
tract.
However,
most
strongly
cells
epithelial
cells.
Based
on
small
intestine
single
cell
RNA-Seq
analysis
different
types,
parp14
was
pronounced
enterocytes
Tuft
Mice
with
body-wide
genetic
deficiency
suffered
exacerbated
colitis.
The
histopathological
large
revealed
increased
infiltration,
Goblet
loss,
erosion.
At
same
time
lower
numbers
viable
bacteria
detected.
bulk
analysis,
transcriptional
signatures
either
missing
or
down-regulated
deficient
mice
These
enriched
genes
related
adhesion,
division
cytoskeletal
rearrangements,
responses.
appears
that
has
potential
functions
regulation
architecture
mice.
Язык: Английский
Zinc-finger PARP proteins ADP-ribosylate alphaviral proteins and are required for interferon-γ–mediated antiviral immunity
Science Advances,
Год журнала:
2025,
Номер
11(5)
Опубликована: Янв. 31, 2025
Viral
manipulation
of
posttranslational
modifications
(PTMs)
is
critical
to
enable
control
over
host
defenses.
Evidence
suggests
that
one
such
PTM,
adenosine
5′-diphosphate
(ADP)–ribosylation,
important
for
viral
replication,
but
the
and
components
involved
are
poorly
understood.
Here,
we
demonstrate
several
human
poly(ADP-ribose)
polymerase
(PARP)
proteins,
including
zinc-finger
domain
containing
PARP7
(TiPARP)
PARP12,
directly
ADP-ribosylate
alphaviral
nonstructural
proteins
(nsPs),
nsP3
nsP4.
These
same
PARP
inhibit
alphavirus
replication
in
a
manner
can
be
antagonized
by
ADP-ribosylhydrolase
activity
virally
encoded
macrodomain.
Last,
find
knockdown
any
three
CCCH
PARPs,
PARP7,
or
enzymatically
inactive
PARP13
(ZAP/ZC3HAV1),
attenuates
antiviral
effects
interferon-γ
on
replication.
Combined
with
evolutionary
analyses,
these
data
suggest
PARPs
share
an
ancestral
function
macrodomains,
indicative
ongoing
conflict
between
ADP-ribosylation
viruses.
Язык: Английский
Application of Pseudoinfectious Viruses in Transient Gene Expression in Mammalian Cells: Combining Efficient Expression with Regulatory Compliance
Biomolecules,
Год журнала:
2025,
Номер
15(2), С. 274 - 274
Опубликована: Фев. 13, 2025
Transient
gene
expression
(TGE)
is
commonly
employed
for
protein
production,
but
its
reliance
on
plasmid
transfection
makes
it
challenging
to
scale
up.
In
this
paper,
an
alternative
TGE
method
presented,
utilizing
pseudoinfectious
alphavirus
as
vector.
Pseudoinfectious
viruses
(PIV)
and
a
replicable
helper
construct
were
derived
from
the
genome
of
Venezuelan
equine
encephalitis
virus.
The
PIV
carries
mutant
capsid
that
prevents
packaging
into
infectious
particles,
while
encodes
wild-type
lacks
other
viral
structural
proteins.
Although
cannot
independently
spread
infection,
their
combination
results
in
increased
titers
cell
cultures,
enabling
easier
scale-up
producing
cultures.
PIV-driven
production
model
outperforms
replicon
vectors
or
simple
vectors.
Another
described
feature
system
modification
immobilized
metal
affinity
chromatography
(IMAC),
allowing
purification
His-tagged
recombinant
proteins
conditioned
medium
presence
substances
can
strip
IMAC
columns.
PIV-based
allows
milligram
quantities
static
without
need
complex
equipment
such
bioreactors,
complies
with
regulatory
requirements
due
distinction
common
viruses.
Язык: Английский
Interferon-induced PARP14-mediated ADP-ribosylation in p62 bodies requires the ubiquitin-proteasome system
The EMBO Journal,
Год журнала:
2025,
Номер
unknown
Опубликована: Апрель 7, 2025
Abstract
Biomolecular
condensates
are
cellular
compartments
without
enveloping
membranes,
enabling
them
to
dynamically
adjust
their
composition
in
response
environmental
changes
through
post-translational
modifications.
Recent
work
has
revealed
that
interferon-induced
ADP-ribosylation
(ADPr),
which
can
be
reversed
by
a
SARS-CoV-2-encoded
hydrolase,
is
enriched
within
condensate.
However,
the
identity
of
condensate
and
responsible
host
ADP-ribosyltransferase
remain
elusive.
Here,
we
demonstrate
interferon
induces
ADPr
transcriptional
activation
PARP14,
requiring
both
physical
presence
catalytic
activity
PARP14
for
formation.
Interferon-induced
colocalizes
with
its
associated
E3
ligase,
DTX3L.
These
PARP14/ADPr
contain
key
components
p62
bodies—including
selective
autophagy
receptor
p62,
binding
partner
NBR1
protein
TAX1BP1,
along
K48-linked
K63-linked
polyubiquitin
chains—but
lack
autophagosome
marker
LC3B.
Knockdown
disrupts
formation
these
condensates.
Importantly,
structures
unaffected
inhibition,
but
depend
on
ubiquitination
proteasome
activity.
Taken
together,
findings
triggers
PARP14-mediated
bodies,
requires
an
active
ubiquitin-proteasome
system.
Язык: Английский
Mutation of a highly conserved isoleucine residue in loop 2 of several β-coronavirus macrodomains indicates that enhanced ADP-ribose binding is detrimental for replication
Journal of Virology,
Год журнала:
2024,
Номер
98(11)
Опубликована: Окт. 10, 2024
ABSTRACT
All
coronaviruses
(CoVs)
encode
for
a
conserved
macrodomain
(Mac1)
located
in
non-structural
protein
3.
Mac1
is
an
ADP-ribosylhydrolase
that
binds
and
hydrolyzes
mono-ADP-ribose
from
target
proteins.
Previous
work
has
shown
important
virus
replication
pathogenesis.
Within
Mac1,
there
are
several
regions
highly
across
CoVs,
including
the
glycine-isoleucine-phenylalanine
motif.
While
we
previously
demonstrated
importance
of
glycine
residue
CoV
pathogenesis,
impact
isoleucine
phenylalanine
residues
remains
unknown.
To
determine
how
biochemical
activities
these
replication,
were
mutated
to
alanine
(I-A/F-A)
both
recombinant
proteins
murine
hepatitis
virus,
Middle
East
respiratory
syndrome
coronavirus
(MERS-CoV),
severe
acute
2
(SARS-CoV-2).
The
F-A
mutant
had
ADP-ribose
binding
and/or
hydrolysis
defects
correlated
with
attenuated
pathogenesis
MERS-CoV
SARS-CoV-2
viruses
cell
culture
mice.
In
contrast,
I-A
normal
enzyme
activity
enhanced
binding.
Despite
only
demonstrating
increased
binding,
mice,
indicating
this
acts
as
gate
controls
efficient
replication.
These
results
highlight
function
provide
unique
insight
into
macrodomains
control
promote
viral
IMPORTANCE
(CoV)
counters
host
ADP-ribosyltransferases
critical
As
such,
potential
therapeutic
CoV-induced
disease.
However,
lack
basic
knowledge
its
pocket
contribute
virological
functions.
We
engineered
mutations
two
(MERS-CoV)
Interestingly,
isoleucine-to-alanine
which
proved
be
detrimental
beneficial
will
development
novel
inhibitors
targeting
could
used
treat
Язык: Английский
PARP14 mediated SQSTM1/p62 cysteine ADP-ribosylation is counteracted by the SARS-CoV-2 macrodomain
bioRxiv (Cold Spring Harbor Laboratory),
Год журнала:
2024,
Номер
unknown
Опубликована: Июль 1, 2024
ABSTRACT
Several
ADP-ribosyltransferases
are
upregulated
during
viral
infections
and
crucial
for
the
cellular
immune
response.
While
interferon-induced
PARP14
ADP-ribosylates
various
substrates,
viruses
such
as
SARS-CoV-2
counteract
this
by
reversing
ADP-ribosylation.
The
exact
mechanism
of
PARP14’s
antiviral
activity
targets
macrodomains
remain
unknown.
Here,
we
observe
that
mono-ADP-ribosylates
selective
autophagy
adaptor
SQSTM1/p62
at
cysteine
residues
113,
289/90,
331
following
interferon
treatment.
This
correlates
with
ADP-ribosylation
cytoplasmic
p62
foci
colocalize
ubiquitin
but
not
LC3,
thereby
distinguishing
them
from
classical
autophagosomes.
Moreover,
macrodomain
effectively
prevented
modification,
suggesting
an
function
ADP-ribosylated
target.
Furthermore,
our
results
indicate
TRIM21
prevents
autophagic
degradation
p62,
identified
may
have
autophagy-independent
roles.
study
contributes
to
understanding
molecular
dynamics
involved
in
host-virus
interactions
highlights
potential
role
regulation
innate
immunity.
Язык: Английский