Nucleic Acids Research, 1992, Vol. 20, No. 21 5587-5592
© 1992
MOLECULAR BIOLOGY |
Isolation of cDNA clones encoding the ß isozyme of human DNA topoisomerase II and localisation of the gene to chromosome 3p24
Imperial Cancer Research Fund Laboratories, Institute of Molecular Medicine, University of Oxford John Radcliffe Hospital, Oxford 0X3 9DU 1Imperial Cancer Research Fund Laboratories, Institute of Molecular Medicine, University of Oxford 44 Lincolns Inn Fields, London WC2A 3PX, UK
* To whom correspondence should be addressed
Received September 1, 1992. Revised October 9, 1992. Accepted October 9, 1992.
Topoisomerases catalyse the interconversion of topological isomers of DNA and have key roles in nucleic acid metabolism. Human cells express two distinct type II topoisomerase isozymes, designatec topoisomerase II
(170 kDa form) and topoisomerase IIß (180 kDa form). We have isolated cDNA clones encoding the ß isozyme from a human B-cell library. The proposed coding region for the topoisomerase IIß protein is 4,863 nucleotides long and would encode i polypeptide with a calculated M, of 182,705. The predicted topoisomerase IIß protein sequence show; striking similarity (72% identical residues) to that of the human isozyme, and homology to topoisomerase II proteins from Drosophila, yeast and bacteria. Region of greatest amino acid sequence divergence lie at the extreme N-terminus and over a C-terminal domain comprising –25% of the total protein. We have quantified the level of topoisomerase IIß mRNA in a panel of human tumour cell lines of different origin using an RNase protection assay, and compared the level to that of topoisomerase II mRNA. Topoisomerase IIß mRNA was expressed inhaemopoietic, epithelial and fibroblast cell lines, although of different extents, with U937 cells (promonocytic leukaemia) showing a particularly high level. There was no obvious relationship in terms of level of expression between the topoisomerase II and ß genes. We have localised the gene encoding topoisomerase IIß protein to chromosome 3p24 in the human genome.
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