eLife,
Journal Year:
2024,
Volume and Issue:
unknown
Published: Nov. 6, 2024
DNA
base
lesions,
such
as
incorporation
of
uracil
into
or
mismatches,
can
be
mutagenic
and
toxic
to
replicating
cells.
To
discover
factors
in
repair
genomic
uracil,
we
performed
a
CRISPR
knockout
screen
the
presence
floxuridine,
chemotherapeutic
agent
that
incorporates
fluoro-uracil
DNA.
We
identified
known
factors,
N-glycosylase
(UNG),
unknown
N6-adenosine
methyltransferase,
METTL3,
required
overcome
floxuridine-driven
cytotoxicity.
Visualized
with
immunofluorescence,
product
METTL3
activity,
N6-methyladenosine,
formed
nuclear
foci
cells
treated
floxuridine.
The
observed
N6-methyladenosine
was
embedded
DNA,
called
6mA,
these
results
were
confirmed
using
an
orthogonal
approach,
liquid
chromatography
coupled
tandem
mass
spectrometry
(LC-MS/MS).
6mA
for
lesions
driven
by
additional
damaging
agents,
including
raltitrexed,
gemcitabine,
hydroxyurea.
Our
establish
role
promoting
genome
stability
mammalian
cells,
especially
response
damage.
International Journal of Cancer,
Journal Year:
2025,
Volume and Issue:
unknown
Published: April 12, 2025
Enhancers
are
critical
regulators
of
gene
expression.
Structural
variations
in
cancer
genomes
can
lead
to
enhancer
hijacking,
where
oncogenes
activated
by
mistargeted
activity.
Novel
enhancer-promoter
interactions
may
also
arise
through
chromosomal
rearrangements
that
create
extrachromosomal
DNA
elements.
Additionally,
fusion
proteins
and
other
mutation-induced
alterations
protein
properties
the
aberrant
assembly
into
large
complexes
on
size
scale
0.1-1
μm
termed
onco-condensates.
Transcription
factors
co-activators
accumulate
with
cis-regulatory
elements
these
structures,
driving
oncogenic
programs.
Here,
we
review
current
evidence
how
altered
genome
architecture
macromolecular
result
deregulated
communication.
We
discuss
emerging
strategies
exploit
mechanisms
for
clinical
applications.
DNA
base
lesions,
such
as
incorporation
of
uracil
into
or
mismatches,
can
be
mutagenic
and
toxic
to
replicating
cells.
To
discover
factors
in
repair
genomic
uracil,
we
performed
a
CRISPR
knockout
screen
the
presence
floxuridine,
chemotherapeutic
agent
that
incorporates
fluoro-uracil
DNA.
We
identified
known
factors,
N-glycosylase
(UNG),
unknown
N6-adenosine
methyltransferase,
METTL3,
required
overcome
floxuridine-driven
cytotoxicity.
Visualized
with
immunofluorescence,
product
METTL3
activity,
N6-methyladenosine,
formed
nuclear
foci
cells
treated
floxuridine.
The
observed
N6-methyladenosine
was
embedded
DNA,
called
6mA,
these
results
were
confirmed
using
an
orthogonal
approach,
liquid
chromatography
coupled
tandem
mass
spectrometry
(LC-MS/MS).
6mA
for
lesions
driven
by
additional
damaging
agents,
including
raltitrexed,
gemcitabine,
hydroxyurea.
Our
establish
role
promoting
genome
stability
mammalian
cells,
especially
response
damage.
ACS Sensors,
Journal Year:
2025,
Volume and Issue:
unknown
Published: Feb. 23, 2025
Biomolecular
condensates
are
membraneless
compartments
with
enigmatic
roles
across
intracellular
phenomena.
Intrinsically
disordered
proteins
(IDPs)
often
function
as
condensate
scaffolds,
fueled
by
liquid–liquid
phase
separation
(LLPS)
dynamics.
Intracellular
probing
of
relies
on
live-cell
imaging
IDP-scaffolds
tagged
fluorescent
proteins.
Conformational
heterogeneity
in
IDPs,
however,
renders
them
uniquely
susceptible
to
artifacts
from
tagging.
Probing
epidermal
skin,
we
recently
introduced
genetically-encoded
LLPS-sensors
that
circumvent
the
need
for
molecular-level
tagging
skin
IDPs.
Departing
subcellular
tracking
IDP-scaffolds,
report
assembly
and
liquid-like
dynamics
their
condensates.
Here,
demonstrate
biomolecular
approaches
evolution
tunability
assess
impact
early
late
stages
separation.
Benchmarking
against
scaffold-bound
reporters,
discovered
tunable
ultraweak
scaffold–sensor
interactions
enable
sensitive
innocuous
nascent
established
Our
LLPS-sensitive
tools
pave
way
high-fidelity
IDP-governed
biological
systems.
DNA
base
lesions,
such
as
incorporation
of
uracil
into
or
mismatches,
can
be
mutagenic
and
toxic
to
replicating
cells.
To
discover
factors
in
repair
genomic
uracil,
we
performed
a
CRISPR
knockout
screen
the
presence
floxuridine,
chemotherapeutic
agent
that
incorporates
fluorouracil
DNA.
We
identified
known
factors,
N-glycosylase
(UNG),
unknown
N6-adenosine
methyltransferase,
METTL3,
required
overcome
floxuridine-driven
cytotoxicity.
Visualized
with
immunofluorescence,
product
METTL3
activity,
N6-methyladenosine,
formed
nuclear
foci
cells
treated
floxuridine.
The
observed
N6-methyladenosine
was
embedded
DNA,
called
6mA,
these
results
were
confirmed
using
an
orthogonal
approach,
liquid
chromatography
coupled
tandem
mass
spectrometry.
6mA
for
lesions
driven
by
additional
base-damaging
agents,
including
raltitrexed,
gemcitabine,
hydroxyurea.
Our
establish
role
promoting
genome
stability
mammalian
cells,
especially
response
damage.
