Biochemical complementation studies in vitro of gyrase subunits from different species

FEBS Letters

Roth, Martin¹  Simon, Hannelore²  Zimmer, Christoph² 

[1] Hans-Knöll-Institut für Naturstoff-Forschung, e. V., Beutenbergstraβe 11, D-07745 Jena, Germany;Institut für Molekularbiologie, Friedrich-Schiller-Universität Jena, Winzerlaer Straβe 10, D-07745 Jena, Germany

关键词: DNA gyrase;    Subunit;    Gyrase hybrid;    DNA binding;    DNA cleavage;    Immunology;   

DOI  :  10.1016/0014-5793(95)01007-2

学科分类：生物化学/生物物理

来源: John Wiley & Sons Ltd.

【 摘 要 】

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.

【 授权许可】

Unknown