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Staphylococcus aureus Nuc2 is a functional, surface-attached extracellular nuclease.
[pyomyositis]
Staphylococcus
aureus
is
a
prominent
bacterial
pathogen
that
causes
a
diverse
range
of
acute
and
chronic
infections
.
Recently
,
it
has
been
demonstrated
that
the
secreted
nuclease
(
Nuc
)
enzyme
is
a
virulence
factor
in
multiple
models
of
infection
,
and
in
vivo
expression
of
nuc
has
facilitated
the
development
of
an
infection
imaging
approach
based
on
Nuc-activatable
probes
.
Interestingly
,
S
.
aureus
strains
encode
a
second
nuclease
(
Nuc
2
)
that
has
received
limited
attention
.
With
the
growing
interest
in
bacterial
nucleases
,
we
sought
to
characterize
Nuc
2
in
more
detail
through
localization
,
expression
,
and
biochemical
studies
.
Fluorescence
microscopy
and
alkaline
phosphatase
localization
approaches
using
Nuc
2
-
GFP
and
Nuc
2
-
PhoA
fusions
,
respectively
,
demonstrated
that
Nuc
2
is
membrane
bound
with
the
C-
terminus
facing
the
extracellular
environment
,
indicating
it
is
a
signal-anchored
Type
II
membrane
protein
.
Nuc
2
enzyme
activity
was
detectable
on
the
S
.
aureus
cell
surface
using
a
fluorescence
resonance
energy
transfer
(
FRET
)
assay
,
and
in
time
courses
,
both
nuc
2
transcription
and
enzyme
activity
peaked
in
early
logarithmic
growth
and
declined
in
stationary
phase
.
Using
a
mouse
model
of
S
.
aureus
pyomyositis
,
Nuc
2
activity
was
detected
with
activatable
probes
in
vivo
in
nuc
mutant
strains
,
demonstrating
that
Nuc
2
is
produced
during
infections
.
To
assess
Nuc
2
biochemical
properties
,
the
protein
was
purified
and
found
to
cleave
both
single
-
and
double
-stranded
DNA
,
and
it
exhibited
thermostability
and
calcium
dependence
,
paralleling
the
properties
of
Nuc
.
Purified
Nuc
2
prevented
biofilm
formation
in
vitro
and
modestly
decreased
biomass
in
dispersal
experiments
.
Altogether
,
our
findings
confirm
that
S
.
aureus
encodes
a
second
,
surface-attached
and
functional
DNase
that
is
expressed
during
infections
and
displays
similar
biochemical
properties
to
the
secreted
Nuc
enzyme
.
Diseases
Validation
Diseases presenting
"and in vivo expression of nuc has facilitated the development of an infection imaging approach based on nuc-activatable probes"
symptom
pyomyositis
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