title: The deviant ATP-binding site of the multidrug efflux pump Pdr5 plays an active role in the transport cycle.
abstract: Pdr5 is the founding member of a large subfamily of evolutionarily distinct, clinically important fungal ABC transporters containing a characteristic, deviant ATP-binding site with altered Walker A, Walker B, Signature (C-loop), and Q-loop residues. In contrast to these motifs, the D-loops of the two ATP-binding sites have similar sequences, including a completely conserved aspartate residue. Alanine substitution mutants in the deviant Walker A and Signature motifs retain significant, albeit reduced, ATPase activity and drug resistance. The D-loop residue mutants D340A and D1042A showed a striking reduction in plasma membrane transporter levels. The D1042N mutation localized properly had nearly WT ATPase activity but was defective in transport and was profoundly hypersensitive to Pdr5 substrates. Therefore, there was a strong uncoupling of ATPase activity and drug efflux. Taken together, the properties of the mutants suggest an additional, critical intradomain signaling role for deviant ATP-binding sites.
authors.affiliation: From the Departments of Biology and.
authors.forename: Christopher
authors.initials: C
authors.lastname: Furman
authors.affiliation: From the Departments of Biology and.
authors.forename: Jitender
authors.initials: J
authors.lastname: Mehla
authors.affiliation: From the Departments of Biology and.
authors.forename: Neeti
authors.initials: N
authors.lastname: Ananthaswamy
authors.affiliation: From the Departments of Biology and.
authors.forename: Nidhi
authors.initials: N
authors.lastname: Arya
authors.affiliation: From the Departments of Biology and.
authors.forename: Bridget
authors.initials: B
authors.lastname: Kulesh
authors.affiliation: From the Departments of Biology and.
authors.forename: Ildiko
authors.initials: I
authors.lastname: Kovach
authors.affiliation: Laboratory of Cell Biology, Center for Cancer Research, NCI, National Institutes of Health, Bethesda, Maryland 20892; Chemistry, Catholic University of America, Washington, D. C. 20064.
authors.forename: Suresh V
authors.initials: SV
authors.lastname: Ambudkar
authors.affiliation: From the Departments of Biology and. Electronic address: golin@cua.edu.
authors.forename: John
authors.initials: J
authors.lastname: Golin
mesh_terms: D018528:ATP-Binding Cassette Transporters / Q000235:genetics / Q000378:metabolism*; D000251:Adenosine Triphosphatases / Q000235:genetics / Q000378:metabolism*; D019943:Amino Acid Substitution; D001665:Binding Sites; D001693:Biological Transport, Active / Q000502:physiology; D026141:Drug Resistance, Multiple, Fungal / Q000502:physiology*; D020125:Mutation, Missense; D017433:Protein Structure, Secondary; D017434:Protein Structure, Tertiary; D012441:Saccharomyces cerevisiae / Q000201:enzymology* / Q000235:genetics; D029701:Saccharomyces cerevisiae Proteins / Q000235:genetics / Q000378:metabolism*
chemical_list: D018528:ATP-Binding Cassette Transporters; C092438:PDR5 protein, S cerevisiae; D029701:Saccharomyces cerevisiae Proteins; D000251:Adenosine Triphosphatases
keywords: ABC Transporter; ATPases; Drug Transport; Membrane Proteins; Multidrug Transporters
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