Eukaryotic topoisomerase II preferentially cleaves alternating purine-pyrimidine repeats

J. R. Spitzner, I. K. Chung, M. T. Muller

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124 Citations (Scopus)

Abstract

Alternating purine-pyrimidine sequences (RY repeats) demonstrate considerable homology to the consensus sequence for vertebrate topoisomerase II (Spitzner and Muller (1988) Nucleic Acids Res. 16: 1533-1556). This is shown below and positions that can match are underscored. (R is purine, Y is pyrimidine, K is G or T.) Topoisomerase II cleavage reactions were performed (in the absence of inhibitors) on a plasmid containing a 54 base RY repeat and the single strong cleavage site mapped to the RY repeat. Analysis of this DNA on sequencing gels showed that the enzyme cleaved a number of sites, all within the 54 base pair RY repeat. Topoisomerase II also made clustered cleavages within other RY repeats that were examined. Quantitative analysis of homology to the consensus sequence, as measured by the match of a site to a matrix of base proportions from the consensus data base (the matrix mean), showed that both the locations and the frequencies of cleavage sites within RY repeats were proportional to homology scores. However, topoisomerase II cleaved RY repeats preferentially in comparison to non-RY sites with similar homology scores. The activity of the enzyme at RY repeats appears to be proportional to the length of the repeat; additionally, GT, AC and AT repeats were better substrates for cleavage than GC repeats.

Original languageEnglish
Pages (from-to)1-11
Number of pages11
JournalNucleic acids research
Volume18
Issue number1
DOIs
Publication statusPublished - 1990 Jan 11

Bibliographical note

Funding Information:
This work was supported by Public Health Service Grant GM31640 from the NIH. The authors are grateful to the following individuals who provided plasmids: L. Johnson (mouse thymidylate synthetase intron 5 clone), E. Vanin (human globin gene region), R.R. Sinden (pCGTA-C). We thank R.R. Sinden for critical reading of the paper and for helpful suggestions.

All Science Journal Classification (ASJC) codes

  • Genetics

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