The initiation of SV40 (simian virus 40) DNA replication requires the co-operative interactions between the viral Tag (large T-antigen), RPA (replication protein A) and Pol (DNA polymerase α-primase) on the template DNA. Binding interfaces mapped on these enzymes and expressed as peptides competed with the mutual interactions of the native proteins. Prevention of the genuine interactions was accomplished only prior to the primer synthesis step and blocked the assembly of a productive initiation complex. Once the complex was engaged in the synthesis of an RNA primer and its extension, the interfering effects of the peptides ceased, suggesting a stable association of the replication factors during the initiation phase. Specific antibodies were still able to disrupt preformed interactions and inhibited primer synthesis and extension activities, underlining the crucial role of specific protein–protein contacts during the entire initiation process.
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Research Article|
September 25 2007
Timed interactions between viral and cellular replication factors during the initiation of SV40 in vitro DNA replication
Poonam Taneja;
Poonam Taneja
1
*Department of Biological Sciences, Vanderbilt University, 465 21st Avenue South, Nashville, TN 37235, U.S.A.
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Heinz-Peter Nasheuer;
Heinz-Peter Nasheuer
†Department of Biochemistry, National University of Ireland, Galway, University Road, Galway, Ireland
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Hella Hartmann;
Hella Hartmann
2
‡Leibniz Institute for Age Research, Fritz Lipmann Institute (formerly Institute for Molecular Biotechnology), Beutenbergstrasse 11, 07745 Jena, Germany
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Frank Grosse;
Frank Grosse
‡Leibniz Institute for Age Research, Fritz Lipmann Institute (formerly Institute for Molecular Biotechnology), Beutenbergstrasse 11, 07745 Jena, Germany
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Ellen Fanning;
Ellen Fanning
*Department of Biological Sciences, Vanderbilt University, 465 21st Avenue South, Nashville, TN 37235, U.S.A.
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Klaus Weisshart
Klaus Weisshart
3
‡Leibniz Institute for Age Research, Fritz Lipmann Institute (formerly Institute for Molecular Biotechnology), Beutenbergstrasse 11, 07745 Jena, Germany
3To whom correspondence should be sent. Present address: Carl Zeiss MicroImaging GmbH, Carl-Zeiss-Promenade 10, 07745 Jena, Germany (email weisshart@zeiss.de).
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Publisher: Portland Press Ltd
Received:
June 14 2007
Revision Received:
July 23 2007
Accepted:
August 01 2007
Accepted Manuscript online:
August 01 2007
Online ISSN: 1470-8728
Print ISSN: 0264-6021
© The Authors Journal compilation © 2007 Biochemical Society
2007
Biochem J (2007) 407 (2): 313–320.
Article history
Received:
June 14 2007
Revision Received:
July 23 2007
Accepted:
August 01 2007
Accepted Manuscript online:
August 01 2007
Citation
Poonam Taneja, Heinz-Peter Nasheuer, Hella Hartmann, Frank Grosse, Ellen Fanning, Klaus Weisshart; Timed interactions between viral and cellular replication factors during the initiation of SV40 in vitro DNA replication. Biochem J 15 October 2007; 407 (2): 313–320. doi: https://doi.org/10.1042/BJ20070794
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