title: Escherichia coli topoisomerase I is an iron and zinc binding protein.
abstract: Escherichia coli topoisomerase I (TopA) cleaves and rejoins one strand of double-stranded DNA to relax the negatively supercoiled DNA. Structurally, TopA contains an N-terminal catalytic fragment and a C-terminal zinc-binding region that is required for relaxation of the negatively supercoiled DNA. Here we report that E. coli TopA is an iron and zinc binding protein. The UV-Vis absorption measurements and metal content analyses reveal that TopA purified from E. coli cells grown in the rich LB medium contains both iron and zinc. However, TopA purified from E. coli cells grown in the M9 minimal medium has negligible amounts of zinc or iron and no topoisomerase activity. Nevertheless, supplement of exogenous zinc or iron in E. coli cells grown in the M9 minimal medium produces the zinc- or iron-bound TopA, respectively. Whereas the zinc-bound TopA is fully active to relax the negatively supercoiled DNA, the iron-bound TopA has little or no enzyme activity. Furthermore, excess iron in the M9 minimal medium is able to compete with the zinc binding in TopA in E. coli cells and attenuate the topoisomerase activity, suggesting that E. coli TopA may be modulated by iron and zinc binding in vivo.
authors.affiliation: Laboratory of Molecular Medicine, Wenzhou Medical College, Wenzhou 325035, Zhejiang, People's Republic of China.
authors.forename: Jianxin
authors.initials: J
authors.lastname: Lu
authors.forename: Wu
authors.initials: W
authors.lastname: Wang
authors.forename: Guoqiang
authors.initials: G
authors.lastname: Tan
authors.forename: Aaron P
authors.initials: AP
authors.lastname: Landry
authors.forename: Peng
authors.initials: P
authors.lastname: Yi
authors.forename: Fan
authors.initials: F
authors.lastname: Si
authors.forename: Yaguang
authors.initials: Y
authors.lastname: Ren
authors.forename: Huangen
authors.initials: H
authors.lastname: Ding
mesh_terms: D001665:Binding Sites; D004264:DNA Topoisomerases, Type I / Q000302:isolation & purification / Q000378:metabolism*; D004926:Escherichia coli / Q000166:cytology / Q000201:enzymology*; D007501:Iron / Q000378:metabolism*; D013056:Spectrophotometry, Ultraviolet; D015032:Zinc / Q000378:metabolism*
chemical_list: D007501:Iron; D004264:DNA Topoisomerases, Type I; D015032:Zinc