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Mol Cell Biol, June 1998, p. 3552-3562, Vol. 18, No. 6
The Institute of Physical and Chemical
Research,
Received 23 June 1997/Returned for modification 15 December
1997/Accepted 2 March 1998
DNA polymerase
0270-7306/98/$04.00+0
Copyright © 1998, American Society for Microbiology. All rights reserved.
The Second-Largest Subunit of the Mouse DNA
Polymerase
-Primase Complex Facilitates Both Production and Nuclear
Translocation of the Catalytic Subunit of DNA Polymerase
-primase is a replication enzyme necessary for
DNA replication in all eukaryotes examined so far. Mouse DNA polymerase
is made up of four subunits, the largest of which is the catalytic
subunit with a molecular mass of 180 kDa (p180). This subunit exists as
a tight complex with the second-largest subunit (p68), whose
physiological role has remained unclear up until now. We set out to
characterize these subunits individually or in combination by using a
cDNA expression system in cultured mammalian cells. Coexpression of p68
markedly increased the protein level of p180, with the result that
ectopically generated DNA polymerase activity was dramatically
increased. Immunofluorescence analysis showed that while either singly
expressed p180 or p68 was localized in the cytoplasm, cotransfection of
both subunits resulted in colocalization in the nucleus. We identified
a putative nuclear localization signal for p180 (residues 1419 to 1437)
and found that interaction with p68 is essential for p180 to
translocate into the nucleus. These results indicate that association
of p180 with p68 is important for both protein synthesis of p180 and
translocation into the nucleus, implying that p68 plays a pivotal role
in the newly synthesized DNA polymerase
complex.
*
Corresponding author. Mailing address: Institute for
Molecular and Cellular Biology, Osaka University, 1-3 Yamada-oka,
Suita, Osaka 565, Japan. Phone: 81-6-879-7975. Fax:
81-6-877-9382. E-mail: fhanaoka{at}imcb.osaka-u.ac.jp.
Mol Cell Biol, June 1998, p. 3552-3562, Vol. 18, No. 6
0270-7306/98/$04.00+0
Copyright © 1998, American Society for Microbiology. All rights reserved.
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