title: A Mycobacterium smegmatis gyrase B specific monoclonal antibody reveals association of gyrase A and B subunits in the cell.
abstract: DNA gyrase is a unique topoisomerase, which plays important roles in macromolecular events like DNA replication, transcription and genetic recombination. In this study a high affinity monoclonal antibody to the gyrase B (GyrB) subunit of Mycobacterium smegmatis was characterized, which did not cross-react with either the Escherichia coli GyrB subunit or with GyrB subunits from other mycobacterial species. The antibody recognized an epitope in the N-terminus, novobiocin-binding domain of GyrB. Immunoprecipitation of gyrase from M. smegmatis cell lysate revealed an association, mediated by ionic interactions, of gyrase A and GyrB subunits in the cell. This antibody is a valuable tool for structure-function analysis and immunocytological studies of mycobacterial DNA gyrase.
authors.affiliation: Department of Microbiology and Cell Biology, Indian Institute of Science, Bangalore 560 012, India.
authors.forename: U H
authors.initials: UH
authors.lastname: Manjunatha
authors.forename: B P
authors.initials: BP
authors.lastname: Somesh
authors.forename: V
authors.initials: V
authors.lastname: Nagaraja
authors.forename: S S
authors.initials: SS
authors.lastname: Visweswariah
mesh_terms: D000595:Amino Acid Sequence; D000911:Antibodies, Monoclonal / Q000276:immunology*; D027081:DNA Gyrase; D004250:DNA Topoisomerases, Type II / Q000737:chemistry / Q000235:genetics / Q000276:immunology* / Q000378:metabolism*; D018604:Epitope Mapping; D008969:Molecular Sequence Data; D020102:Mycobacterium smegmatis / Q000201:enzymology* / Q000276:immunology; D009675:Novobiocin / Q000378:metabolism; D016415:Sequence Alignment; D020349:Surface Plasmon Resonance / Q000379:methods
chemical_list: D000911:Antibodies, Monoclonal; D009675:Novobiocin; D027081:DNA Gyrase; D004250:DNA Topoisomerases, Type II