Novel Macrolide-Specific ABC-Type Efflux Transporter in Escherichia coli
Open Access
- 1 October 2001
- journal article
- Published by American Society for Microbiology in Journal of Bacteriology
- Vol. 183 (19), 5639-5644
- https://doi.org/10.1128/jb.183.19.5639-5644.2001
Abstract
In the Escherichia coli genome, five putative open reading frame (ORF) clusters, mdlAB , ybjYZ , yddA , yojHI , and yhiH , have been assumed to be possible genes for ABC drug efflux transporters (I. T. Paulsen, M. K. Sliwinski, and M. H. Saier, Jr., J. Mol. Biol. 277:573–592, 1998). We cloned all of these ORFs in multicopy plasmids and investigated the drug resistance of drug-supersensitive host cells lacking constitutive multidrug efflux transporter genes acrAB . Among them, only ybjYZ gave significant erythromycin resistance and significantly decreased the accumulation of [ 14 C]erythromycin. Therefore, ybjYZ was renamed macAB (macrolide-specific ABC-type efflux carrier). Plasmids carrying both the macA and - B genes conferred resistance against macrolides composed of 14- and 15-membered lactones but no or weak resistance against 16-membered ones. Neither of the two genes produced resistance alone. The DNA sequence suggests that MacB is an integral membrane protein with four transmembrane segments and one nucleotide-binding domain, while MacA belongs to a membrane fusion protein (MFP) family with a signal-like sequence at its N terminus. The expression of the histidine-tagged proteins confirmed that MacB is an integral membrane protein and MacA is a peripheral membrane protein. In addition, MacAB required TolC for its function in a way similar to that of most of the MFP-dependent transporters in E. coli . MacB is thus a novel ABC-type macrolide efflux transporter which functions by cooperating with the MFP MacA and the multifunctional outer membrane channel TolC. This is the first case of an experimentally identified ABC antibiotic efflux transporter in gram-negative organisms.Keywords
This publication has 34 references indexed in Scilit:
- Overexpression of the Response Regulator evgA of the Two-Component Signal Transduction System Modulates Multidrug Resistance Conferred by Multidrug Resistance TransportersJournal of Bacteriology, 2001
- Complete DNA sequence of a serogroup A strain of Neisseria meningitidis Z2491Nature, 2000
- Complete Genome Sequence of Neisseria meningitidis Serogroup B Strain MC58Science, 2000
- ABC-ATPases, adaptable energy generators fuelling transmembrane movement of a variety of molecules in organisms from bacteria to humansJournal of Molecular Biology, 1999
- A plasmid that encodes three genes for resistance to macrolide antibiotics inStaphylococcus aureusFEMS Microbiology Letters, 1998
- Microbial genome analyses: global comparisons of transport capabilities based on phylogenies, bioenergetics and substrate specificitiesJournal of Molecular Biology, 1998
- The Complete Genome Sequence of Escherichia coli K-12Science, 1997
- Topology prediction of membrane proteinsProtein Science, 1996
- BIOCHEMISTRY OF MULTIDRUG RESISTANCE MEDIATED BY THE MULTIDRUG TRANSPORTERAnnual Review of Biochemistry, 1993
- A new subfamily of bacterial ABC‐type transport systems catalyzing export of drugs and carbohydratesProtein Science, 1992