bioRxiv (Cold Spring Harbor Laboratory),
Journal Year:
2023,
Volume and Issue:
unknown
Published: Oct. 13, 2023
SUMMARY
In
multinuclear
and
multicellular
filamentous
fungi
little
is
known
about
how
mRNAs
encoding
secreted
enzymes
are
transcribed
localized
spatiotemporally.
To
better
understand
this
process
we
visualized
mRNA
GlaA,
a
glucoamylase
in
large
amounts
by
the
industrial
fungus
Aspergillus
oryzae
,
living
cells
MS2
system.
We
found
that
glaA
was
significantly
near
hyphal
tip
septum,
which
sites
of
protein
secretion.
also
revealed
exhibits
long-range
dynamics
vicinity
endoplasmic
reticulum
(ER)
manner
dependent
on
microtubule
motor
proteins
kinesin-1
kinesin-3,
but
independent
early
endosomes.
Moreover,
elucidated
regulatory
mechanisms
stress
granules
processing
bodies
were
different
under
high
temperature
ER
stress.
Collectively,
study
uncovers
dynamic
mechanism
secretory
enzyme
fungi.
Microbiological Research,
Journal Year:
2024,
Volume and Issue:
282, P. 127653 - 127653
Published: Feb. 23, 2024
In
multinuclear
and
multicellular
filamentous
fungi
little
is
known
about
how
mRNAs
encoding
secreted
enzymes
are
transcribed
localized
spatiotemporally.
To
better
understand
this
process
we
analyzed
mRNA
GlaA,
a
glucoamylase
in
large
amounts
by
the
industrial
fungus
Aspergillus
oryzae,
MS2
system,
which
can
be
visualized
living
cells.
We
found
that
glaA
was
significantly
near
hyphal
tip
septum,
sites
of
protein
secretion,
polarity-dependent
expression
localization
manners.
also
revealed
exhibits
long-range
dynamics
vicinity
endoplasmic
reticulum
(ER)
manner
dependent
on
microtubule
motor
proteins
kinesin-1
kinesin-3,
but
independent
early
endosomes.
Moreover,
elucidated
although
to
stress
granules
(SGs)
processing
bodies
(PBs)
under
high
temperature,
not
seen
ER
stress,
suggesting
there
different
regulatory
mechanisms
SG
PB
temperature
stress.
Collectively,
study
uncovers
dynamic
mechanism
secretory
enzyme
fungi.
PLoS Biology,
Journal Year:
2024,
Volume and Issue:
22(3), P. e3002526 - e3002526
Published: March 1, 2024
Live
imaging
of
RNA
molecules
constitutes
an
invaluable
means
to
track
the
dynamics
mRNAs,
but
live
in
Caenorhabditis
elegans
has
been
difficult
achieve.
Endogenous
transcripts
have
observed
nuclei,
endogenous
mRNAs
not
detected
cytoplasm,
and
functional
generated.
Here,
we
adapted
methods
visualize
mRNA
embryonic
cells.
We
tagged
with
MS2
hairpins
3′
untranslated
region
(UTR)
visualized
them
after
adjusting
Coat
Protein
(MCP)
expression.
A
reduced
number
these
accumulates
leading
loss-of-function
phenotypes.
In
addition,
during
epithelial
morphogenesis,
MS2-tagged
for
dlg-1
fail
associate
adherens
junction,
as
untagged,
mRNAs.
These
defects
are
reversed
by
inactivating
nonsense-mediated
decay
pathway.
mutant
phenotypes
rescued,
associates
junction.
data
suggest
that
repeats
can
induce
degradation
RNAs
alter
their
cytoplasmic
distribution.
Although
our
focus
is
expressed
cells
find
this
method
be
applied
other
cell
types
stages.
The Journal of Cell Biology,
Journal Year:
2024,
Volume and Issue:
224(1)
Published: Oct. 14, 2024
Maximizing
cell
survival
under
stress
requires
rapid
and
transient
adjustments
of
RNA
protein
synthesis.
However,
capturing
these
dynamic
changes
at
both
single-cell
level
across
an
organism
has
been
challenging.
Here,
we
developed
a
system
named
MONITTR
(MS2-embedded
mCherry-based
monitoring
transcription)
for
real-time
simultaneous
measurement
nascent
transcripts
endogenous
levels
in
C.
elegans.
Utilizing
this
system,
monitored
the
transcriptional
bursting
fasting-induced
genes
found
that
epidermis
responds
to
fasting
by
modulating
proportion
actively
transcribing
nuclei
kinetics
individual
alleles.
Additionally,
our
findings
revealed
essential
roles
transcription
factors
NHR-49
HLH-30
governing
fasting.
Furthermore,
tracked
dynamics
during
heat-shock
response
ER
unfolded
observed
conditions.
Collectively,
study
provides
foundation
quantitatively
investigating
how
animals
spatiotemporally
modulate
various
physiological
pathological
Development,
Journal Year:
2023,
Volume and Issue:
150(19)
Published: Oct. 1, 2023
The
distribution
of
mRNA
in
tissue
is
determined
by
the
balance
between
transcription
and
decay.
Understanding
control
RNA
decay
during
development
has
been
somewhat
neglected
compared
with
transcriptional
control.
Here,
we
explore
potential
for
to
trigger
rapid
cell-state
transitions
development,
comparing
a
bistable
switch
model
conversion
experimental
evidence
from
different
developmental
systems.
We
also
consider
another
role
large-scale
that
emerged
studies
stress-induced
transitions,
which
removal
unblocks
translation
machinery
prioritise
synthesis
proteins
establish
new
cell
state.
bioRxiv (Cold Spring Harbor Laboratory),
Journal Year:
2023,
Volume and Issue:
unknown
Published: June 13, 2023
Abstract
Live
imaging
of
RNA
molecules
constitutes
an
invaluable
means
to
track
the
dynamics
mRNAs,
but
live
in
Caenorhabditis
elegans
has
been
difficult
achieve.
Endogenous
transcripts
have
observed
nuclei,
endogenous
mRNAs
not
detected
cytoplasm,
and
functional
generated.
Here,
we
adapted
methods
visualize
mRNA
embryonic
epithelial
cells.
We
tagged
with
MS2
hairpins
3’
Untranslated
Region
(UTR)
visualized
them
after
adjusting
Coat
Protein
(MCP)
expression.
A
reduced
number
these
accumulate
leading
loss-of-function
phenotypes.
In
addition,
for
dlg-1
fail
associate
adherens
junction,
as
mRNA.
These
defects
are
reversed
by
inactivating
nonsense-mediated
decay
pathway.
accumulates
associates
mutant
phenotypes
rescued.
data
suggest
that
repeats
can
induce
degradation
targets
alter
cytoplasmic
distribution.
Although
our
focus
is
RNAs
expressed
cells
during
morphogenesis,
this
method
likely
be
applied
other
cell
types
stages.
Summary
statement
An
MS2-MCP
tag
C.
embryos
without
affecting
stability.
bioRxiv (Cold Spring Harbor Laboratory),
Journal Year:
2024,
Volume and Issue:
unknown
Published: Nov. 21, 2024
RNA
localization
and
regulation
are
critical
for
cellular
function,
yet
many
live
imaging
tools
suffer
from
limited
sensitivity
due
to
background
emissions
unbound
probes.
Here,
we
introduce
conditionally
stable
variants
of
MS2
PP7
coat
proteins
(which
name
dMCP
dPCP)
designed
decrease
in
live-cell
imaging.
Using
a
protein
engineering
approach
that
combines
circular
permutation
degron
masking,
generated
dPCP
rapidly
degrade
except
when
bound
cognate
ligands.
These
enhancements
enabled
the
sensitive
visualization
single
mRNA
molecules
undergoing
differential
within
various
sub-compartments
cells.
We
further
demonstrate
dual-color
with
orthogonal
motifs,
allowing
simultaneous
low-background
distinct
species
same
cell.
Overall,
this
work
provides
versatile,
probes
imaging,
which
should
have
broad
utility
biotechnological
utilization
MS2-
PP7-containing
RNAs.
