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PMID: 11544226 Published · ppublish English Journal Article Research Support, Non-U.S. Gov't

Novel macrolide-specific ABC-type efflux transporter in Escherichia coli.

Journal of bacteriology ·Vol. 183 ·No. 19 ·2001-10-00 ·Pages 5639-44

Kobayashi N, Nishino K, Yamaguchi A

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.

MeSH Terms
ATP-Binding Cassette Transporters/chemistry,genetics,metabolism Amino Acid Sequence Anti-Bacterial Agents/pharmacology Bacterial Outer Membrane Proteins/genetics,metabolism Drug Resistance, Microbial Erythromycin/metabolism,pharmacology Escherichia coli/drug effects,metabolism Escherichia coli Proteins Membrane Transport Proteins Microbial Sensitivity Tests Molecular Sequence Data Open Reading Frames/genetics
Chemicals
ATP-Binding Cassette Transporters Anti-Bacterial Agents Bacterial Outer Membrane Proteins Escherichia coli Proteins MacA protein, E coli MacB protein, E coli Membrane Transport Proteins tolC protein, E coli Erythromycin
Authors & Affiliations
3 authors, click to expand affiliations / ORCID
Kobayashi N
Department of Cell Membrane Biology, Institute of Scientific and Industrial Research, Osaka University, Ibaraki, Japan.
Nishino K
Yamaguchi A
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Article Info
Journal
Journal of bacteriology
Abbr.
J Bacteriol
ISSN
0021-9193
DOI
10.1128/JB.183.19.5639-5644.2001
Published
2001-10-00
Pages
5639-44
Language
English
Region
United States
NLM ID
2985120R
PMCID
PMC95455
Subset
IM
External Links
PubMed source DOI: 10.1128/JB.183.19.5639-5644.2001 PMC Full Text View journal details
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