Roles of DNA topoisomerases in simian virus 40 DNA replication in vitro

Liu Yang; M. S WOLD; Joachim J LI; Thomas J Kelly; Leroy F Liu

doi:10.1073/pnas.84.4.950

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Roles of DNA topoisomerases in simian virus 40 DNA replication in vitro

Journal article

Open access

Peer reviewed

Roles of DNA topoisomerases in simian virus 40 DNA replication in vitro

Liu Yang, M. S WOLD, Joachim J LI, Thomas J Kelly and Leroy F Liu

Proceedings of the National Academy of Sciences - PNAS, Vol.84(4), pp.950-954

1987

DOI: 10.1073/pnas.84.4.950

PMID: 3029765

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Abstract

We examined the roles of DNA topoisomerases in the replication of simian virus 40 (SV40) DNA in a cell-free system composed of an extract from HeLa cells supplemented with purified SV40 tumor antigen. When the activities of both topoisomerase I (EC 5.99.1.2) and topoisomerase II (EC 5.99.1.3) in the extract were blocked with specific inhibitors or antibodies, DNA synthesis was decreased by a factor of 15-20. Addition of purified HeLa DNA topoisomerase II to extracts immunologically depleted of both topoisomerases completely restored replication, and the replication products consisted largely of monomeric daughter molecules. Addition of purified HeLa DNA topoisomerase I to depleted extracts restored DNA synthesis, but the primary products were multiply intertwined, catenated daughter molecules. We conclude that DNA topoisomerases have at least two roles in the replication of SV40 DNA. Either topoisomerase I or topoisomerase II is sufficient to provide the unlinking activity necessary for fork propagation during SV40 DNA replication. However, topoisomerase II is uniquely required for the segregation of newly synthesized daughter molecules.

Molecular Genetics

Fundamental and applied biological sciences. Psychology

Biological and medical sciences

Replication

Molecular and cellular biology

Details

Title: Subtitle: Roles of DNA topoisomerases in simian virus 40 DNA replication in vitro
Creators: Liu Yang - Johns Hopkins University School of Medicine
M. S WOLD - Johns Hopkins univ. school medicine, dep. biological chemistry, Baltimore MD 21205, United States
Joachim J LI - Johns Hopkins univ. school medicine, dep. biological chemistry, Baltimore MD 21205, United States
Thomas J Kelly - Johns Hopkins univ. school medicine, dep. biological chemistry, Baltimore MD 21205, United States
Leroy F Liu - Johns Hopkins univ. school medicine, dep. biological chemistry, Baltimore MD 21205, United States
Resource Type: Journal article
Publication Details: Proceedings of the National Academy of Sciences - PNAS, Vol.84(4), pp.950-954
DOI: 10.1073/pnas.84.4.950
PMID: 3029765
NLM abbreviation: Proc Natl Acad Sci U S A
ISSN: 0027-8424
eISSN: 1091-6490
Publisher: National Academy of Sciences; Washington, DC
Language: English
Date published: 1987
Academic Unit: Radiation Oncology; Biochemistry and Molecular Biology
Record Identifier: 9984024419002771

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