A single inactivating amino acid change in the SARS-CoV-2 NSP3 Mac1 domain attenuates viral replication in vivo
PLoS Pathogens,
Год журнала:
2023,
Номер
19(8), С. e1011614 - e1011614
Опубликована: Авг. 31, 2023
Despite
unprecedented
efforts,
our
therapeutic
arsenal
against
SARS-CoV-2
remains
limited.
The
conserved
macrodomain
1
(Mac1)
in
NSP3
is
an
enzyme
exhibiting
ADP-ribosylhydrolase
activity
and
a
possible
drug
target.
To
determine
the
role
of
Mac1
catalytic
viral
replication,
we
generated
recombinant
viruses
replicons
encoding
catalytically
inactive
domain
by
mutating
critical
asparagine
active
site.
While
substitution
to
alanine
(N40A)
reduced
~10-fold,
mutations
aspartic
acid
(N40D)
~100-fold
relative
wild-type.
Importantly,
N40A
mutation
rendered
unstable
vitro
lowered
expression
levels
bacterial
mammalian
cells.
When
incorporated
into
molecular
clones,
N40D
mutant
only
modestly
affected
fitness
immortalized
cell
lines,
but
replication
human
airway
organoids
10-fold.
In
mice,
replicated
at
>1000-fold
lower
compared
wild-type
virus
while
inducing
robust
interferon
response;
all
animals
infected
with
survived
infection.
Our
data
validate
as
promising
target
develop
antivirals.
Язык: Английский
Exploring pyrazolines as potential inhibitors of NSP3-macrodomain of SARS-CoV-2: synthesis and in silico analysis
Scientific Reports,
Год журнала:
2025,
Номер
15(1)
Опубликована: Янв. 4, 2025
COVID-19
has
proved
to
be
a
global
health
crisis
during
the
pandemic,
and
emerging
JN.1
variant
is
potential
threat.
Therefore,
finding
alternative
antivirals
of
utmost
priority.
In
current
report,
we
present
synthesis
new
anti-viral
pyrazoline
compounds.
Here
report
chemical
scheme
where
β-aryl
β-anilino
ketones
react
with
phenyl
hydrazine
in
potassium
hydroxide
give
corresponding
3,5-diarylpyrazoline.
The
protocol
applicable
variety
β-amino
tolerates
several
functional
groups.
This
method
efficient
proceeds
regioselectivity
since
β-Anilino
group
acts
as
protecting
for
alkenes
chalcones.
We
identified
NSP3-microdomain
(Mac-1)
SARS-CoV-2
putative
target
newly
synthesized
triaryl-2-pyrazoline
molecular
dynamics
simulation-based
free
energy
estimation
suggests
compounds
7a,
7d,
7
g,
7i,
7k,
L
promising
Mac-1
inhibitors.
detailed
structural
inspection
MD
simulation
trajectories
sheds
light
on
involved
Mac-1.
data
presented
here
expected
guide
design
development
better
anti-SARS-CoV-2
therapies.
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
PARPs and ADP-Ribosylation in Chronic Inflammation: A Focus on Macrophages
Pathogens,
Год журнала:
2023,
Номер
12(7), С. 964 - 964
Опубликована: Июль 23, 2023
Aberrant
adenosine
diphosphate-ribose
(ADP)-ribosylation
of
proteins
and
nucleic
acids
is
associated
with
multiple
disease
processes
such
as
infections
chronic
inflammatory
diseases.
The
poly(ADP-ribose)
polymerase
(PARP)/ADP-ribosyltransferase
(ART)
family
members
promote
mono-
or
poly-ADP-ribosylation.
Although
evidence
has
linked
PARPs/ARTs
macrophages
in
the
context
inflammation,
underlying
mechanisms
remain
incompletely
understood.
This
review
provides
an
overview
literature
focusing
on
roles
PARP1/ARTD1,
PARP7/ARTD14,
PARP9/ARTD9,
PARP14/ARTD8
macrophages.
regulate
changes
during
not
only
via
catalytic
modifications
but
also
non-catalytic
mechanisms.
Untangling
complex
mechanisms,
by
which
modulate
macrophage
phenotype,
providing
molecular
bases
for
development
new
therapeutics
require
implementation
innovative
technologies.
Язык: Английский
A single inactivating amino acid change in the SARS-CoV-2 NSP3 Mac1 domain attenuates viral replication and pathogenesisin vivo
bioRxiv (Cold Spring Harbor Laboratory),
Год журнала:
2023,
Номер
unknown
Опубликована: Апрель 18, 2023
Despite
unprecedented
efforts,
our
therapeutic
arsenal
against
SARS-CoV-2
remains
limited.
The
conserved
macrodomain
1
(Mac1)
in
NSP3
is
an
enzyme
exhibiting
ADP-ribosylhydrolase
activity
and
a
possible
drug
target.
To
determine
the
potential
of
Mac1
inhibition,
we
generated
recombinant
viruses
replicons
encoding
catalytically
inactive
domain
by
mutating
critical
asparagine
active
site.
While
substitution
to
alanine
(N40A)
reduced
catalytic
~10-fold,
mutations
aspartic
acid
(N40D)
~100-fold
relative
wildtype.
Importantly,
N40A
mutation
rendered
unstable
Язык: Английский
CoVEx: SARS-CoV-2 Mutation Explorer for genomic surveillance
Infection Genetics and Evolution,
Год журнала:
2023,
Номер
116, С. 105521 - 105521
Опубликована: Окт. 28, 2023
Effective
management
of
emerging
diseases
relies
on
timely
pathogen
identification
and
monitoring.
The
emergence
COVID-19
in
December
2019,
rapidly
evolved
into
a
global
pandemic,
with
millions
cases
deaths
reported
worldwide.
accumulation
SARS-CoV-2
genomes
provided
unprecedented
opportunities
for
studying
the
virus's
evolutionary
dynamics,
understanding
impact
mutations,
identifying
Variants
Interest
(VOIs)
Concern
(VOCs).
During
health
systems
faced
challenges
promptly
detecting
such
variants
notifying.
To
facilitate
continuous
monitoring
various
initiatives
open-source
pipelines
have
been
established.
However,
these
platforms
often
lack
integration
conducting
user
sequence
analysis
comparing
it
publicly
data
like
GISAID.
Here,
we
present
CoVEx,
an
easy-to-use
tool
analyzing
visualizing
variant
sequences
obtained
using
Illumina
sequencing
technology.
CoVEx
integrates
quality
control,
alignment,
genome
annotation,
lineage
designation,
mutation
tools.
Implemented
Python,
also
has
explorer
feature
that
generates
interactive
graphs
summarising
identified
mutations
intuitive
manner.
Similarly,
leverages
Outbreak.info
package
to
create
heatmaps
highlighting
associated
designated
Pangolin
lineages.
Furthermore,
by
profiles
against
GISAID
data,
offers
valuable
insights
prevalence
distribution
We
validated
raw
(n
=
108)
demonstrated
its
accuracy
assembling
predicting
Nextclade
Pango
Notably,
revealed
previously
unreported
mutation,
ORF1a:I2501T,
within
Costa
Rica
GN.1
lineage.
This
finding
highlights
CoVEx's
capability
identify
novel
different
lineages,
providing
information
researchers
public
decision
makers.
documentation
are
freely
available
GitLab:
https://gitlab.com/CNCA_CeNAT/covex.
Язык: Английский