Journal of Experimental Botany,
Год журнала:
2024,
Номер
75(17), С. 5412 - 5427
Опубликована: Фев. 29, 2024
Abstract
Macroautophagy
is
often
quantified
by
live
imaging
of
autophagosomes
labeled
with
fluorescently
tagged
ATG8
protein
(FP–ATG8)
in
Arabidopsis
thaliana.
The
particles
are
then
counted
single
focal
planes.
This
approach
may
lead
to
inaccurate
results
as
the
actual
3D
distribution
not
taken
into
account
and
appropriate
sampling
Z-direction
performed.
To
overcome
this
issue,
we
developed
a
workflow
consisting
immunolabeling
an
anti-ATG8
antibody
followed
stereological
image
analysis
using
optical
disector
Cavalieri
principle.
Our
protocol
specifically
recognized
epidermal
cells
root.
Since
recognizes
multiple
AtATG8
isoforms,
were
able
detect
higher
number
immunolabeled
than
FP–AtATG8e
marker,
that
most
probably
does
recognize
all
cell.
per
tissue
volume
positively
correlated
intensity
autophagy
induction.
Compared
quantification
maximum
projections,
methods
present
given
accuracy.
novel
provides
powerful
toolkit
for
unbiased
reproducible
offers
convenient
alternative
standard
FP–ATG8
markers.
bioRxiv (Cold Spring Harbor Laboratory),
Год журнала:
2024,
Номер
unknown
Опубликована: Апрель 23, 2024
Vacuoles
are
essential
for
cellular
metabolism,
growth,
and
the
maintenance
of
internal
turgor
pressure.
They
sequester
lytic
enzymes,
ions,
secondary
metabolites
that,
if
leaked
into
cytosol,
could
lead
to
cell
death.
Despite
their
pivotal
roles,
quality
control
pathways
that
safeguard
vacuolar
integrity
remained
elusive
in
plants.
Here,
we
discovered
a
conserved
(VQC)
pathway
is
activated
upon
wall
damage
pressure
dependent
manner.
Cell
perturbations
induce
distinct
modification
-
ATG8ylation
on
membrane
(tonoplast)
regulated
by
V-ATPase
ATG8
conjugation
machinery.
Genetic
disruption
tonoplast
impairs
integrity,
leading
Together,
our
findings
reveal
homeostatic
preserves
damage.
International Journal of Molecular Sciences,
Год журнала:
2024,
Номер
25(20), С. 11160 - 11160
Опубликована: Окт. 17, 2024
Autophagosome
(AP)-lysosome/vacuole
fusion
is
one
of
the
hallmarks
macroautophagy.
Membrane
features
and
changes
during
process
have
mostly
been
described
using
two-dimensional
(2D)
models
with
AP
lysosome/vacuole.
The
outer
membrane
(OM)
a
closed
mature
has
suggested
to
fuse
lysosomal/vacuolar
membrane.
However,
descriptions
in
some
studies
for
fusion-related
issues
are
questionable
or
incomplete.
correct
APs
lysosomes/vacuoles
prerequisite
describing
process.
We
searched
literature
representative
AP-related
structures
based
on
electron
microscopy
(EM)
graphs
both
animal
yeast
cells
re-evaluated
findings.
also
summarized
main
2D
AP-lysosome/vacuole
literature.
used
three-dimensional
(3D)
characterize
known
unknown
after
most
plausible
models.
actual
situation
more
complex,
since
multiple
lysosomes
may
same
mammalian
cells,
vacuole
mutant
phagophores
(unclosed
APs)
lysosomes/vacuoles.
This
review
discusses
highly
dynamic
(phagophore)-lysosome/vacuole
fusion.
resulting
information
will
improve
understanding
direct
future
research
regeneration.
Frontiers in Plant Science,
Год журнала:
2023,
Номер
14
Опубликована: Март 15, 2023
Phosphatidylinositol
3-phosphate
(PI3P)
is
a
signaling
phospholipid
that
play
key
role
in
endomembrane
trafficking,
specifically
autophagy
and
endosomal
trafficking.
However,
the
mechanisms
underlying
contribution
of
PI3P
downstream
effectors
to
plant
remain
unknown.
Known
for
Arabidopsis
thaliana
include
ATG18A
(Autophagy-related
18A)
FYVE2
(Fab1p,
YOTB,
Vac1p,
EEA1
2),
which
are
implicated
autophagosome
biogenesis.
Here,
we
report
FYVE3,
paralog
plant-specific
FYVE2,
plays
FYVE2-dependent
autophagy.
Using
yeast
two-hybrid
bimolecular
fluorescence
complementation
assays,
determined
FYVE3
protein
was
associated
with
autophagic
machinery
containing
by
interacting
ATG8
isoforms.
The
transported
vacuole,
vacuolar
delivery
relies
on
biosynthesis
canonical
machinery.
Whereas
fyve3
mutation
alone
barely
affects
flux,
it
suppresses
defective
fyve2
mutants.
Based
molecular
genetics
cell
biological
data,
propose
regulates
FEBS Letters,
Год журнала:
2022,
Номер
596(17), С. 2305 - 2313
Опубликована: Май 20, 2022
Autophagy
fulfills
a
crucial
role
in
plant
cellular
homeostasis
by
recycling
diverse
components
ranging
from
protein
complexes
to
whole
organelles.
cargos
are
shuttled
the
vacuole
for
degradation,
thereby
completing
process.
Canonical
autophagy
requires
lipidation
and
insertion
of
ATG8
proteins
into
double-membrane
structures,
termed
autophagosomes,
which
engulf
cargo
be
degraded.
As
such,
pathway
actively
contributes
intracellular
membrane
trafficking.
Yet,
autophagic
process
is
not
fully
considered
bona
fide
component
canonical
trafficking
pathway.
However,
recent
findings
have
started
pinpoint
interconnection
between
classical
pathways
autophagy.
This
review
details
latest
advances
our
comprehension
interplay
these
two
pathways.
Understanding
overlap
important
illuminate
inner
workings
both
cells.
Journal of Experimental Botany,
Год журнала:
2024,
Номер
75(17), С. 5412 - 5427
Опубликована: Фев. 29, 2024
Abstract
Macroautophagy
is
often
quantified
by
live
imaging
of
autophagosomes
labeled
with
fluorescently
tagged
ATG8
protein
(FP–ATG8)
in
Arabidopsis
thaliana.
The
particles
are
then
counted
single
focal
planes.
This
approach
may
lead
to
inaccurate
results
as
the
actual
3D
distribution
not
taken
into
account
and
appropriate
sampling
Z-direction
performed.
To
overcome
this
issue,
we
developed
a
workflow
consisting
immunolabeling
an
anti-ATG8
antibody
followed
stereological
image
analysis
using
optical
disector
Cavalieri
principle.
Our
protocol
specifically
recognized
epidermal
cells
root.
Since
recognizes
multiple
AtATG8
isoforms,
were
able
detect
higher
number
immunolabeled
than
FP–AtATG8e
marker,
that
most
probably
does
recognize
all
cell.
per
tissue
volume
positively
correlated
intensity
autophagy
induction.
Compared
quantification
maximum
projections,
methods
present
given
accuracy.
novel
provides
powerful
toolkit
for
unbiased
reproducible
offers
convenient
alternative
standard
FP–ATG8
markers.
