Mapping the active-site tyrosine of vaccinia virus DNA topoisomerase I

S. Shuman, E. M. Kane, S. G. Morham

Research output: Contribution to journalArticle

50 Citations (Scopus)

Abstract

Site-directed mutagenesis of the vaccinia virus gene encoding a type I DNA topoisomerase implicates Tyr-274 as the active-site residue that forms a covalent adduct with DNA during cycles of DNA-strand breakage and reunion. Replacement of Tyr-274 by phenylalanine results in loss of the ability of the enzyme to relax negatively supercoiled DNA as well as to form the covalent DNA-protein intermediate. Substitution of phenylalanine for tyrosine at nine other sites in the protein has no apparent effect on enzyme activity. Amino acid sequence alignment reveals Tyr-274 to be homologous to Tyr-727 and Tyr-771, respectively, of the type I topoisomerases from Saccharomyces cerevisiae and Saccharomyces pombe; Tyr-727 and Tyr-771 have been shown to represent the active-site tyrosines of those enzymes. Sequence comparison of the active-site regions defines a motif Ser-Lys-Xaa-Xaa-Tyr common to the viral and cellular type I topoisomerases, including the human enzyme.

Original languageEnglish (US)
Pages (from-to)9793-9797
Number of pages5
JournalProceedings of the National Academy of Sciences of the United States of America
Volume86
Issue number24
DOIs
StatePublished - Dec 1 1989

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Type I DNA Topoisomerase
Vaccinia virus
Tyrosine
Catalytic Domain
Enzymes
Phenylalanine
Reunion
Superhelical DNA
Saccharomyces
DNA Adducts
Sequence Alignment
DNA
Site-Directed Mutagenesis
Saccharomyces cerevisiae
Amino Acid Sequence
Proteins
Genes

All Science Journal Classification (ASJC) codes

  • General

Cite this

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Mapping the active-site tyrosine of vaccinia virus DNA topoisomerase I. / Shuman, S.; Kane, E. M.; Morham, S. G.

In: Proceedings of the National Academy of Sciences of the United States of America, Vol. 86, No. 24, 01.12.1989, p. 9793-9797.

Research output: Contribution to journalArticle

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AB - Site-directed mutagenesis of the vaccinia virus gene encoding a type I DNA topoisomerase implicates Tyr-274 as the active-site residue that forms a covalent adduct with DNA during cycles of DNA-strand breakage and reunion. Replacement of Tyr-274 by phenylalanine results in loss of the ability of the enzyme to relax negatively supercoiled DNA as well as to form the covalent DNA-protein intermediate. Substitution of phenylalanine for tyrosine at nine other sites in the protein has no apparent effect on enzyme activity. Amino acid sequence alignment reveals Tyr-274 to be homologous to Tyr-727 and Tyr-771, respectively, of the type I topoisomerases from Saccharomyces cerevisiae and Saccharomyces pombe; Tyr-727 and Tyr-771 have been shown to represent the active-site tyrosines of those enzymes. Sequence comparison of the active-site regions defines a motif Ser-Lys-Xaa-Xaa-Tyr common to the viral and cellular type I topoisomerases, including the human enzyme.

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