Journal of the American Chemical Society,
Год журнала:
2024,
Номер
146(27), С. 18722 - 18729
Опубликована: Июнь 29, 2024
Methylation,
a
widely
occurring
natural
modification
serving
diverse
regulatory
and
structural
functions,
is
carried
out
by
myriad
of
S-adenosyl-l-methionine
(AdoMet)-dependent
methyltransferases
(MTases).
The
AdoMet
cofactor
produced
from
l-methionine
(Met)
ATP
family
multimeric
methionine
adenosyltransferases
(MAT).
To
advance
mechanistic
functional
studies,
strategies
for
repurposing
the
MAT
MTase
reactions
to
accept
extended
versions
transferable
group
corresponding
precursors
have
been
exploited.
Here,
we
used
structure-guided
engineering
mouse
MAT2A
enable
biocatalytic
production
an
analogue,
Ado-6-azide,
synthetic
S-(6-azidohex-2-ynyl)-l-homocysteine
(N3-Met).
Three
engineered
variants
showed
catalytic
proficiency
with
analogues
supported
DNA
derivatization
in
cascade
M.TaqI
variant
DNMT1
both
absence
presence
competing
Met.
We
then
installed
two
as
MAT2A-DNMT1
cascades
embryonic
stem
cells
using
CRISPR-Cas
genome
editing.
resulting
cell
lines
maintained
normal
viability
methylation
levels
Dnmt1-dependent
azide
tags
upon
exposure
N3-Met
physiological
This
first
time
demonstrates
genetically
stable
system
biosynthetic
which
enables
mild
metabolic
labeling
DNMT-specific
methylome
live
mammalian
cells.
Journal of Virology,
Год журнала:
2024,
Номер
98(5)
Опубликована: Апрель 19, 2024
Alphaherpesvirus
pseudorabies
virus
(PRV)
causes
severe
economic
losses
to
the
global
pig
industry
and
has
garnered
increasing
attention
due
its
broad
host
range
including
humans.
PRV
developed
a
variety
of
strategies
antagonize
antiviral
innate
immunity.
However,
underlying
mechanisms
have
not
been
fully
elucidated.
In
our
previous
work,
we
demonstrated
that
non-muscle
myosin
heavy
chain
IIA
(NMHC-IIA),
multifunctional
cytoskeleton
protein,
attenuates
immune
responses
triggered
by
RNA
viruses.
current
study,
reported
previously
unrecognized
role
NMHC-IIA
in
counteracting
PRV-induced
cyclic
GMP-AMP
synthase
(cGAS)-dependent
type
I
interferon
(IFN-I)
production.
Mechanistically,
infection
led
an
elevation
NMHC-IIA,
strengthening
interaction
between
poly
(ADP-ribose)
polymerase
1
(PARP1)
cGAS.
This
impeded
cGAS
recognition
DNA
hindered
downstream
signaling
activation.
Conversely,
inhibition
Blebbistatin
enhanced
resistance
proliferation
both
Journal of the American Chemical Society,
Год журнала:
2024,
Номер
146(27), С. 18722 - 18729
Опубликована: Июнь 29, 2024
Methylation,
a
widely
occurring
natural
modification
serving
diverse
regulatory
and
structural
functions,
is
carried
out
by
myriad
of
S-adenosyl-l-methionine
(AdoMet)-dependent
methyltransferases
(MTases).
The
AdoMet
cofactor
produced
from
l-methionine
(Met)
ATP
family
multimeric
methionine
adenosyltransferases
(MAT).
To
advance
mechanistic
functional
studies,
strategies
for
repurposing
the
MAT
MTase
reactions
to
accept
extended
versions
transferable
group
corresponding
precursors
have
been
exploited.
Here,
we
used
structure-guided
engineering
mouse
MAT2A
enable
biocatalytic
production
an
analogue,
Ado-6-azide,
synthetic
S-(6-azidohex-2-ynyl)-l-homocysteine
(N3-Met).
Three
engineered
variants
showed
catalytic
proficiency
with
analogues
supported
DNA
derivatization
in
cascade
M.TaqI
variant
DNMT1
both
absence
presence
competing
Met.
We
then
installed
two
as
MAT2A-DNMT1
cascades
embryonic
stem
cells
using
CRISPR-Cas
genome
editing.
resulting
cell
lines
maintained
normal
viability
methylation
levels
Dnmt1-dependent
azide
tags
upon
exposure
N3-Met
physiological
This
first
time
demonstrates
genetically
stable
system
biosynthetic
which
enables
mild
metabolic
labeling
DNMT-specific
methylome
live
mammalian
cells.
