Proceedings of the National Academy of Sciences,
Год журнала:
2025,
Номер
122(14)
Опубликована: Апрель 1, 2025
Lenacapavir
(GS-6207;
LEN)
is
a
potent
HIV-1
capsid
inhibitor
approved
for
treating
multidrug-resistant
infection.
LEN
binds
to
hydrophobic
pocket
between
neighboring
(CA)
proteins
in
hexamers
and
stabilizes
the
lattice,
but
its
effect
on
capsids
not
fully
understood.
Here,
we
labeled
with
green
fluorescent
protein
fused
CA
(GFP-CA)
or
fluid-phase
GFP
content
marker
(cmGFP)
assess
LEN’s
impact
capsids.
cores
GFP-CA,
cmGFP,
could
be
immunostained
an
anti-GFP
antibody
were
less
sensitive
capsid-binding
host
restriction
factor
MX2,
demonstrating
that
GFP-CA
incorporated
into
lattice
stability,
whereas
cmGFP
indicator
of
core
integrity.
treatment
isolated
resulted
dose-dependent
loss
signal
while
preserving
signal,
indicating
disrupts
integrity
lattice.
In
contrast,
PF-3450074
(PF74)
induced
Electron
microscopy
LEN-
PF74-treated
viral
revealed
frequent
breakage
at
narrow
end
other
morphological
changes.
Our
results
suggest
does
prevent
nuclear
envelope
docking
inhibits
import
without
PF74
blocks
by
inhibiting
cores,
highlighting
their
different
mechanisms
inhibition.
Journal of Molecular Cell Biology,
Год журнала:
2022,
Номер
14(11)
Опубликована: Окт. 31, 2022
ABSTRACT
HIV
integration
occurs
in
chromatin
sites
that
favor
the
release
of
high
levels
viral
progeny;
alternatively,
virus
is
also
able
to
discreetly
coexist
with
host.
The
infection
perturbs
cellular
environment
inducing
remodelling
nuclear
landscape.
Indeed,
HIV-1
triggers
clustering
host
factor
CPSF6,
but
underlying
mechanism
poorly
understood.
Our
data
indicate
usurps
a
recently
discovered
biological
phenomenon,
called
liquid–liquid
phase
separation,
hijack
cell.
We
observed
CPSF6
clusters
as
part
HIV-induced
membraneless
organelles
(HIV-1
MLOs)
macrophages,
one
main
target
cell
types.
describe
MLOs
follow
phase-separation
rules
and
represent
functional
biomolecular
condensates.
highlight
hubs
reverse
transcription,
while
double-stranded
DNA,
once
formed,
rapidly
migrates
outside
these
structures.
Transcription-competent
proviruses
localize
near
LEDGF-abundant
regions,
known
be
active
sites.
Therefore,
orchestrate
events
prior
step
create
favorable
for
replication.
This
study
uncovers
single
host–viral
complexes
their
landscape,
which
markedly
restructured
by
HIV-1.
Abstract
HIV‐1
uses
inositol
hexakisphosphate
(IP6)
to
build
a
metastable
capsid
capable
of
delivering
its
genome
into
the
host
nucleus.
Here,
we
show
that
viruses
are
unable
package
IP6
lack
protection
and
detected
by
innate
immunity,
resulting
in
activation
an
antiviral
state
inhibits
infection.
Disrupting
enrichment
results
defective
capsids
trigger
cytokine
chemokine
responses
during
infection
both
primary
macrophages
T‐cell
lines.
Restoring
with
single
mutation
rescues
ability
infect
cells
without
being
detected.
Using
combination
mutants
CRISPR‐derived
knockout
cell
lines
for
RNA
DNA
sensors,
immune
sensing
is
dependent
upon
cGAS–STING
axis
independent
detection.
Sensing
requires
synthesis
viral
prevented
reverse
transcriptase
inhibitors
or
active‐site
mutation.
These
demonstrate
required
can
successfully
transit
avoid
sensing.
Cell Reports,
Год журнала:
2024,
Номер
43(3), С. 113941 - 113941
Опубликована: Март 1, 2024
Resting
CD4
T
cells
resist
productive
HIV-1
infection.
The
HIV-2/simian
immunodeficiency
virus
protein
viral
accessory
X
(Vpx)
renders
these
permissive
to
infection,
presumably
by
alleviating
blocks
at
cytoplasmic
reverse
transcription
and
subsequent
nuclear
import
of
reverse-transcription/pre-integration
complexes
(RTC/PICs).
Here,
spatial
analyses
using
quantitative
imaging
techniques
reveal
that
capsids
containing
RTC/PICs
are
readily
imported
into
the
nucleus,
recruit
host
dependency
factor
CPSF6,
translocate
speckles
in
resting
cells.
Reverse
transcription,
however,
remains
incomplete,
impeding
proviral
integration
gene
expression.
Vpx
or
pharmacological
inhibition
deoxynucleotide
triphosphohydrolase
(dNTPase)
activity
restriction
SAM
domain
HD
domain-containing
1
(SAMHD1)
increases
levels
reverse-transcribed
cDNA
facilitates
integration.
Nuclear
intranuclear
transport
therefore
do
not
pose
important
cells,
limitation
SAMHD1's
dNTPase
constitutes
main
pre-integration
block
ABSTRACT
A
critical
determinant
for
early
post-entry
events,
the
HIV-1
capsid
(CA)
protein
forms
conical
core
when
it
rearranges
around
dimeric
RNA
genome
and
associated
viral
proteins.
Although
mutations
in
CA
have
been
reported
to
alter
innate
immune
sensing
of
HIV-1,
a
direct
link
between
stability
nucleic
acids
has
not
established.
Herein,
we
assessed
how
manipulating
lattice
through
chemical
genetic
approaches
affects
recognition
HIV-1.
We
found
that
destabilization
resulted
potent
reverse
transcription
products
per
se
does
completely
block
transcription.
Surprisingly,
due
combined
effects
enhanced
defects
nuclear
entry,
two
separate
mutants
form
hyperstable
cores
induced
more
potently
than
destabilizing
mutations.
At
low
concentrations
allowed
accumulation
products,
CA-targeting
compounds
GS-CA1
lenacapavir
measurably
impacted
cells
modestly
HIV.
Interestingly,
activation
observed
with
viruses
containing
unstable
was
abolished
by
doses
lenacapavir.
Innate
both
dependent
on
cGAS-STING
DNA-sensing
pathway
Overall,
our
findings
demonstrate
are
finely
balanced
support
minimize
cGAS-STING-mediated
resulting
DNA.
IMPORTANCE
In
particles,
proteins
enzymes
encased
proteinaceous
composed
protein.
altering
this
orthogonal
impacts
induction
responses.
Specifically,
decreasing
results
but
genomic
RNA,
cGAS-STING-dependent
manner.
The
recently
developed
inhibitors
Unexpectedly,
increased
levels
cytosolic
cDNA,
also
type
I
interferon-mediated
immunity.
Our
suggest
exposure
cytosol
host
cells.
Proceedings of the National Academy of Sciences,
Год журнала:
2025,
Номер
122(14)
Опубликована: Апрель 1, 2025
Lenacapavir
(GS-6207;
LEN)
is
a
potent
HIV-1
capsid
inhibitor
approved
for
treating
multidrug-resistant
infection.
LEN
binds
to
hydrophobic
pocket
between
neighboring
(CA)
proteins
in
hexamers
and
stabilizes
the
lattice,
but
its
effect
on
capsids
not
fully
understood.
Here,
we
labeled
with
green
fluorescent
protein
fused
CA
(GFP-CA)
or
fluid-phase
GFP
content
marker
(cmGFP)
assess
LEN’s
impact
capsids.
cores
GFP-CA,
cmGFP,
could
be
immunostained
an
anti-GFP
antibody
were
less
sensitive
capsid-binding
host
restriction
factor
MX2,
demonstrating
that
GFP-CA
incorporated
into
lattice
stability,
whereas
cmGFP
indicator
of
core
integrity.
treatment
isolated
resulted
dose-dependent
loss
signal
while
preserving
signal,
indicating
disrupts
integrity
lattice.
In
contrast,
PF-3450074
(PF74)
induced
Electron
microscopy
LEN-
PF74-treated
viral
revealed
frequent
breakage
at
narrow
end
other
morphological
changes.
Our
results
suggest
does
prevent
nuclear
envelope
docking
inhibits
import
without
PF74
blocks
by
inhibiting
cores,
highlighting
their
different
mechanisms
inhibition.