title: The TetA(K) tetracycline/H(+) antiporter from Staphylococcus aureus: mutagenesis and functional analysis of motif C.
abstract: Conserved motif C, identified within members of the major facilitator superfamily (MFS) of transport proteins that mediate drug export, was examined in the tetracycline resistance efflux protein TetA(K) from Staphylococcus aureus; motif C is contained within transmembrane segment 5. Using site-directed mutagenesis, the importance of the conserved glycine (G151, G155, G159, and G160) and proline (P156) residues within this motif was investigated. Over 40 individual amino acid replacements were introduced; however, only alanine and serine substitutions for glycine at G151, G155, and G160 were found to retain significant levels of tetracycline resistance and transport activity in cells expressing mutant proteins. Notably, P156 and G159 appear to be crucial, as amino acid replacements at these positions either significantly reduced or abolished tetracycline/H(+) activity. The highly conserved nature of motif C and its distribution throughout drug exporters imply that the residues of motif C play a similar role in all MFS proteins that function as antiporters.
authors.affiliation: School of Biological Sciences, University of Sydney, Sydney, New South Wales 2006, Australia.
authors.forename: S L
authors.initials: SL
authors.lastname: Ginn
authors.forename: M H
authors.initials: MH
authors.lastname: Brown
authors.forename: R A
authors.initials: RA
authors.lastname: Skurray
mesh_terms: D020816:Amino Acid Motifs / Q000235:genetics*; D017920:Antiporters / Q000235:genetics* / Q000378:metabolism*; D001426:Bacterial Proteins / Q000235:genetics* / Q000378:metabolism*; D002462:Cell Membrane / Q000378:metabolism; D016297:Mutagenesis, Site-Directed; D013211:Staphylococcus aureus / Q000187:drug effects / Q000235:genetics* / Q000378:metabolism; D013752:Tetracycline / Q000378:metabolism; D013753:Tetracycline Resistance / Q000235:genetics*
chemical_list: D017920:Antiporters; D001426:Bacterial Proteins; C079051:tetA protein, Bacteria; D013752:Tetracycline