Species specificity of simian virus 40 DNA replication in vitro requires multiple functions of human DNA polymerase alpha

The Journal of Biological Chemistry
R W P SmithHeinz-Peter Nasheuer

Abstract

Human cell extracts support the replication of SV40 DNA, whereas mouse cell extracts do not. Species specificity is determined at the level of initiation of DNA replication, and it was previously found that this requires the large subunit, p180, of DNA polymerase alpha-primase to be of human origin. Furthermore, a functional interaction between SV40 large T antigen (TAg) and p180 is essential for viral DNA replication. In this study we determined that the N-terminal regions of human p180, which contain the TAg-binding sites, can be replaced with those of murine origin without losing the ability to support SV40 DNA replication in vitro. The same substitutions do not prevent SV40 TAg from stimulating the activity of DNA polymerase alpha-primase on single-stranded DNA in the presence of replication protein A. Furthermore, biophysical studies show that the interactions of human and murine DNA polymerase alpha-primase with SV40 TAg are of a similar magnitude. These studies strongly suggest that requirement of SV40 DNA replication for human DNA polymerase alpha depends neither on the TAg-binding site being of human origin nor on the strength of the binary interaction between SV40 TAg and DNA polymerase alpha-primase but rather on seq...Continue Reading

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Citations

Dec 31, 2003·Experimental Gerontology·Vinod K Srivastava, David L Busbee
Apr 24, 2003·European Journal of Biochemistry·Richard W P Smith, Heinz-Peter Nasheuer
Nov 21, 2008·Proceedings of the National Academy of Sciences of the United States of America·Tina T SaxowskyPaul W Doetsch
Mar 20, 2009·Journal of Virology·Cathal MahonWilliam R Folk
May 6, 2011·Journal of Virology·Irina TikhanovichHeinz Peter Nasheuer
Jun 12, 2010·Virology Journal·Daraporn Pittayakhajonwut, Peter C Angeletti
Jan 21, 2004·Journal of Molecular Recognition : JMR·Rebecca L Rich, David G Myszka
Jan 30, 2008·Current Protocols in Cell Biology·Christopher B Buck, Cynthia D Thompson

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