title: Characterization of P55, a multidrug efflux pump in Mycobacterium bovis and Mycobacterium tuberculosis.
abstract: The Mycobacterium bovis P55 gene, located downstream from the gene that encodes the immunogenic lipoprotein P27, has been characterized. The gene was identical to the open reading frame of the Rv1410c gene in the genome of Mycobacterium tuberculosis H37Rv, annotated as a probable drug efflux protein. Genes similar to P55 were present in all species of the M. tuberculosis complex and other mycobacteria such as Mycobacterium leprae and Mycobacterium avium. By Western blotting, P55 was located in the membrane fraction of M. bovis. When transformed into Mycobacterium smegmatis after cloning, P55 conferred aminoglycoside and tetracycline resistance. The levels of resistance to streptomycin and tetracycline conferred by P55 were decreased in the presence of the protonophore carbonyl cyanide m-chlorophenylhydrazone and the pump inhibitors verapamil and reserpine. M. smegmatis cells expressing the plasmid-encoded P55 accumulated less tetracycline than the control cells. We conclude that P55 is a membrane protein implicated in aminoglycoside and tetracycline efflux in mycobacteria.
authors.affiliation: Departamento de Microbiología, Facultad de Medicina, Universidad de Zaragoza, 50009-Zaragoza, Spain.
authors.forename: P E
authors.initials: PE
authors.lastname: Silva
authors.forename: F
authors.initials: F
authors.lastname: Bigi
authors.forename: M P
authors.initials: MP
authors.lastname: Santangelo
authors.forename: M I
authors.initials: MI
authors.lastname: Romano
authors.forename: C
authors.initials: C
authors.lastname: Martín
authors.forename: A
authors.initials: A
authors.lastname: Cataldi
authors.forename: J A
authors.initials: JA
authors.lastname: Aínsa
mesh_terms: D020816:Amino Acid Motifs; D000900:Anti-Bacterial Agents / Q000494:pharmacology; D001426:Bacterial Proteins / Q000302:isolation & purification / Q000378:metabolism*; D002352:Carrier Proteins / Q000037:antagonists & inhibitors / Q000302:isolation & purification / Q000378:metabolism*; D016208:Databases, Factual; D018432:Drug Resistance, Multiple; D008074:Lipoproteins / Q000502:physiology; D026901:Membrane Transport Proteins; D008826:Microbial Sensitivity Tests; D009163:Mycobacterium bovis / Q000187:drug effects / Q000378:metabolism*; D009169:Mycobacterium tuberculosis / Q000187:drug effects / Q000378:metabolism*; D013307:Streptomycin / Q000494:pharmacology; D013347:Subcellular Fractions; D013753:Tetracycline Resistance / Q000502:physiology*; D013754:Tetracyclines; D014700:Verapamil / Q000494:pharmacology
chemical_list: D000900:Anti-Bacterial Agents; D001426:Bacterial Proteins; D002352:Carrier Proteins; D008074:Lipoproteins; D026901:Membrane Transport Proteins; C110121:P27 protein, Mycobacterium; C425143:P55 protein, Mycobacterium; D013754:Tetracyclines; D014700:Verapamil; D013307:Streptomycin