Life Science Alliance,
Journal Year:
2024,
Volume and Issue:
8(1), P. e202402899 - e202402899
Published: Dec. 2, 2024
Multisubunit
protein
complexes
are
central
to
many
cellular
processes,
and
studying
their
activities
structures
in
vitro
requires
reconstitution
via
recombinant
expression
purification.
Obtaining
targets
at
sufficient
purity
scale
typically
involves
screening
several
variants
hosts.
Existing
cloning
strategies
enable
co-expression
but
often
time-consuming,
labor-intensive,
host-specific,
or
involve
error-prone
steps.
We
present
a
novel
vector
set
assembly
strategy
overcome
these
limitations,
enabling
construct
generation
for
multisubunit
single
step.
This
modular
system
can
be
extended
additional
hosts
include
new
tags.
demonstrate
its
utility
by
constructing
vectors
structural
maintenance
of
chromosomes
various
hosts,
streamlining
workflows,
improving
productivity.
Nucleic Acids Research,
Journal Year:
2024,
Volume and Issue:
unknown
Published: Nov. 26, 2024
Bacteria
encode
a
wide
array
of
immune
systems
to
protect
themselves
against
ubiquitous
bacteriophages
and
foreign
DNA
elements.
While
these
systems'
molecular
mechanisms
are
becoming
increasingly
well
known,
their
regulation
remains
poorly
understood.
Here,
we
show
that
an
system-associated
transcriptional
repressor
the
wHTH-WYL-WCX
family,
CapW,
directly
binds
single-stranded
sense
damage
activate
expression
its
associated
system.
We
CapW
mediates
increased
reporter
gene
in
response
host
cell.
by-products
repair
through
WYL
domain,
causing
conformational
change
releases
protein
from
double-stranded
DNA.
In
Escherichia
coli
CBASS
system
with
integrated
capW
gene,
find
CapW-mediated
activation
is
important
for
this
system's
ability
prevent
induction
λ
prophage.
Overall,
our
data
reveal
WYL-domain
repressors,
provide
example
how
bacteria
can
balance
protective
benefits
carrying
anti-phage
inherent
risk
aberrant
activation.
Life Science Alliance,
Journal Year:
2024,
Volume and Issue:
8(1), P. e202402899 - e202402899
Published: Dec. 2, 2024
Multisubunit
protein
complexes
are
central
to
many
cellular
processes,
and
studying
their
activities
structures
in
vitro
requires
reconstitution
via
recombinant
expression
purification.
Obtaining
targets
at
sufficient
purity
scale
typically
involves
screening
several
variants
hosts.
Existing
cloning
strategies
enable
co-expression
but
often
time-consuming,
labor-intensive,
host-specific,
or
involve
error-prone
steps.
We
present
a
novel
vector
set
assembly
strategy
overcome
these
limitations,
enabling
construct
generation
for
multisubunit
single
step.
This
modular
system
can
be
extended
additional
hosts
include
new
tags.
demonstrate
its
utility
by
constructing
vectors
structural
maintenance
of
chromosomes
various
hosts,
streamlining
workflows,
improving
productivity.
