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Journal of Virology, February 2001, p. 1751-1760, Vol. 75, No. 4
Abteilung Biochemie, Institut für
Molekulare Biotechnologie e.V., D-07745 Jena,1
and Genzentrum, Ludwig-Maximilians-Universität
München, D-81377 Munich,2 Germany
Received 20 July 2000/Accepted 17 November 2000
DNA polymerase
0022-538X/01/$04.00+0 DOI: 10.1128/JVI.75.4.1751-1760.2001
Copyright © 2001, American Society for Microbiology. All rights reserved.
Different Regions of Primase Subunit p48 Control
Mouse Polyomavirus and Simian Virus 40 DNA Replication In
Vitro


-primase (pol-prim), a complex consisting of four
subunits, is the major species-specific factor for mouse polyomavirus
(PyV) and simian virus 40 (SV40) DNA replication. Although p48 is the
most conserved subunit of pol-prim, it is required for in vitro PyV DNA
replication but can inhibit cell-free SV40 DNA replication. Production
of chimeric human-mouse p48 revealed that different regions of p48 are
involved in supporting PyV DNA replication and inhibiting SV40 DNA
replication. The N and C-terminal parts of p48 do not have
species-specific functions in cell-free PyV DNA replication, but the
central part (amino acids [aa] 129 to 320) controls PyV DNA
replication in vitro. However, PyV T antigen physically binds to mouse,
human, and chimeric pol-prim complexes independently, whether they
support PyV DNA replication or not. In contrast to the PyV system, the
inhibitory effects of mouse p48 on SV40 DNA replication are mediated by
N- and C-terminal regions of p48. Thus, a chimeric p48 containing human
aa 1 to 128, mouse aa 129 to 320, and human aa 321 to 418 is active in both PyV and SV40 DNA replication in vitro.
*
Corresponding author. Mailing address: Institut
für Molekulare Biotechnologie e.V., Abt. Biochemie,
Beutenbergstr. 11, D-07745 Jena, Germany. Phone: 49-3641-656290. Fax:
49-3641-656288. E-mail: nasheuer{at}imb-jena.de.
Present address: Institut für Biochemie, Universität
Erlangen-Nürnberg, D-91054 Erlangen, Germany.
Present address: Carl Zeiss Jena GmbH, D-07745 Jena, Germany.
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