Studies on the initiation and elongation reactions in the simian virus 40 DNA replication system Journal Article


Authors: Matsumoto, T.; Eki, T.; Hurwitz, J.
Article Title: Studies on the initiation and elongation reactions in the simian virus 40 DNA replication system
Abstract: The synthesis of oligoribonucleotides by DNA primase in the presence of duplex DNA containing the simian virus 40 (SV40) origin of replication was examined. Small RNA chains (10-15 nucleotides) were synthesized in the presence of the four common ribonucleoside triphosphates, SV40 large tumor antigen (T antigen), the human DNA polymerase a (pol α)-DNA primase complex, the human single-stranded DNA-binding protein (HSSB), and topoisomerase I isolated from HeLa cells. The DNA primase-catalyzed reaction showed an absolute requirement for T antigen, HSSB, and pol α. The requirement for HSSB was not satisfied by other SSBs that can support the T-antigen-catalyzed unwinding of DNA containing the SV40 origin of replication. Oligoribonucleotide synthesis occurred with a lag that paralleled the lag observed in DNA synthesis. These results indicate that the specificity for the HSSB in the SV40 replication reaction is due to the pol α-primase-mediated synthesis of the Okazaki fragments. In contrast to this specificity, the elongation of Okazaki fragments can be catalyzed by a variety of different DNA polymerases, including high levels of pol α, the polymerase δ holoenzyme, T4 polymerase holoenzyme, the Escherichia coli polymerase III holoenzyme, and other polymerases. These observations suggest that leading-strand synthesis in the in vitro SV40 replication system can be nonspecific.
Keywords: dna-binding proteins; nonhuman; dna polymerase; dna replication; hela cells; kinetics; escherichia coli; simian virus 40; dna, viral; virus dna; dna-directed dna polymerase; dna protein complex; dna primase; oligoribonucleotides; simiae; dna polymerase ii; rna nucleotidyltransferases; simian virus; human; priority journal; article; support, u.s. gov't, p.h.s.; protein-dna complexes; leading-strand synthesis; dna polymerase α-dna primase complex; oligoribonucleotide primer; t-phages
Journal Title: Proceedings of the National Academy of Sciences of the United States of America
Volume: 87
Issue: 24
ISSN: 0027-8424
Publisher: National Academy of Sciences  
Date Published: 1990-12-01
Start Page: 9712
End Page: 9716
Language: English
DOI: 10.1073/pnas.87.24.9712
PUBMED: 2175912
PROVIDER: scopus
PMCID: PMC55243
DOI/URL:
Notes: Article -- Export Date: 27 January 2020 -- Source: Scopus
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  1. Jerard Hurwitz
    206 Hurwitz