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Kaposi's sarcoma-associated herpesvirus ORF6 gene is essential in viral lytic replication.
[primary effusion lymphoma]
Kaposi
's
sarcoma
associated
herpesvirus
(
KSHV
)
is
associated
with
Kaposis
's
sarcoma
(
KS
)
,
primary
effusion
lymphoma
and
multicentric
Castleman
's
disease
.
KSHV
encodes
at
least
8
open
reading
frames
(
ORFs
)
that
play
important
roles
in
its
lytic
DNA
replication
.
Among
which
,
ORF
6
of
KSHV
encodes
an
ssDNA
binding
protein
that
has
been
proved
to
participate
in
origin-dependent
DNA
replication
in
transient
assays
.
To
define
further
the
function
of
ORF
6
in
the
virus
life
cycle
,
we
constructed
a
recombinant
virus
genome
with
a
large
deletion
within
the
ORF
6
locus
by
using
a
bacterial
artificial
chromosome
(
BAC
)
system
.
Stable
293
T
cells
carrying
the
BAC
36
(
wild
type
)
and
BACΔ
6
genomes
were
generated
.
When
monolayers
of
293
T
-BAC
36
and
293
T
-BACΔ
6
cells
were
induced
with
12
-
O-
tetradecanoylphorbol-
13
-
acetate
(
TPA
)
and
sodium
butyrate
,
infectious
virus
was
detected
from
the
293
T
-BAC
36
cell
supernatants
only
and
not
from
the
293
T
-
BACΔ
6
cell
supernatants
.
DNA
synthesis
was
defective
in
293
T
-BACΔ
6
cells
.
Expression
of
ORF
6
in
trans
in
BACΔ
6
-
containing
cells
was
able
to
rescue
both
defects
.
Our
results
provide
genetic
evidence
that
ORF
6
is
essential
for
KSHV
lytic
replication
.
The
stable
293
T
cells
carrying
the
BAC
36
and
BACΔ
6
genomes
could
be
used
as
tools
to
investigate
the
detailed
functions
of
ORF
6
in
the
lytic
replication
of
KSHV
.
