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PMID: 6267594 Published · ppublish English Journal Article Research Support, Non-U.S. Gov't Research Support, U.S. Gov't, P.H.S.

Eukaryotic DNA topoisomerases: two forms of type I DNA topoisomerases from HeLa cell nuclei.

Proceedings of the National Academy of Sciences of the United States of America ·Vol. 78 ·No. 6 ·1981-06-00 ·Pages 3487-91

Liu LF, Miller KG

Abstract

Two type I DNA topoisomerases have been purified to homogeneity from the nuclei of HeLa cells. One topoisomerase has a peptide molecular weight of 100,000 and the other, a molecular weight of 67,000. Several lines of evidence indicate that these two topoisomerases are closely related, (a) Both exhibit similar enzymatic activities on DNA. (b) The chromatographic properties of the two topoisomerases during purification are similar. (c) Mild proteolysis of the purified molecular weight 100,000 topoisomerase in vitro generates a group of protein bands of molecular weight approximately 67,000, and these bands retain topoisomerase activity. (d) The peptides formed by partial proteolysis of the 67,000 topoisomerase in the presence of NaDodSO4 form a subset of those produced from the 100,000 enzyme. The 100,000 topoisomerase is the major type I enzyme in the cell. The 67,000 topoisomerase, which may be identical to the previously identified "nicking-closing" enzyme [Champoux, J. J. & Dulbecco, R. (1972) Proc. Natl. Acad. Sci. USA 69, 143-146], is probably formed by proteolysis of the 100,000 enzyme.

MeSH Terms
Cell Nucleus/enzymology DNA Topoisomerases, Type I/isolation & purification,metabolism Eukaryotic Cells/enzymology HeLa Cells/enzymology Humans Molecular Weight Peptide Fragments/analysis Trypsin/metabolism
Chemicals
Peptide Fragments Trypsin DNA Topoisomerases, Type I
Authors & Affiliations
2 authors, click to expand affiliations / ORCID
Liu L F
Miller K G
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33 references, click to expand
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Article Info
Journal
Proceedings of the National Academy of Sciences of the United States of America
Abbr.
Proc Natl Acad Sci U S A
ISSN
0027-8424
DOI
10.1073/pnas.78.6.3487
Published
1981-06-00
Pages
3487-91
Language
English
Region
United States
NLM ID
7505876
PMCID
PMC319594
Subset
IM
Grants
NIGMS NIH HHS · GM 27731 · United States
External Links
PubMed source DOI: 10.1073/pnas.78.6.3487 PMC Full Text View journal details
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