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Biochemical complementation studies in vitro of gyrase subunits from different species
Author(s) -
Simon Hannelore,
Roth Martin,
Zimmer Christoph
Publication year - 1995
Publication title -
febs letters
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.593
H-Index - 257
eISSN - 1873-3468
pISSN - 0014-5793
DOI - 10.1016/0014-5793(95)01007-2
Subject(s) - dna gyrase , dna supercoil , dna , pbr322 , circular bacterial chromosome , heterologous , biology , complementation , protein subunit , cleavage (geology) , escherichia coli , biochemistry , plasmid , microbiology and biotechnology , topoisomerase , recbcd , nuclease , recombinant dna , dna repair , dna replication , phenotype , gene , paleontology , fracture (geology)
To investigate the functional equivalence of DNA gyrase subunits from different bacterial sources hybrid enzymes were formed using purified A and B subunits from three species of Streptomycetes, E. coli and B. subtilis . The activity of gyrase hybrids composed of heterologous gyr A and gyr B proteins and of the gyrases containing homologous subunits was characterized by binding studies and a cleavage assay with two different DNA fragments. Likewise the enzyme activity was monitored by the super‐coiling and relaxation assay with pBR322 DNA. We found that cleavage reactions are largely determined by the source of the gyr B subunits whereas DNA supercoiling and relaxation reactions of pBR322 catalyzed by DNA gyrase are limited to a combination of homologous A and B subunits or of heterologous A and B subunits from the taxonomically related bacteria Streptomycetes .