Generating Symmetry in the Asymmetric ATP-binding Cassette (ABC) Transporter Pdr5 from Saccharomyces cerevisiae

Gupta, Rakeshkumar P.; Kueppers, Petra; Hanekop, Nils; Schmitt, Lutz

doi:10.1074/jbc.m114.553065

Cited by 34 publications

(44 citation statements)

References 35 publications

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“…Thus, their observations indicate that Snq2p and Pdr5p share common substrates, but also have specificities. The relationships between Pdr5p structure and activity have been extensively studied, 20,[28][29][30][31][32] but the structure-function relationships of Snq2p have not been reported. Therefore, it is interesting and important to compare Snq2p and Pdr5p for improving our understanding of ABC transporters.…”

Section: Discussionmentioning

confidence: 99%

Multidrug resistance transporters Snq2p and Pdr5p mediate caffeine efflux in Saccharomyces cerevisiae

Tsujimoto

Shimizu

Otake

et al. 2015

Bioscience, Biotechnology, and Biochemistry

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SNQ2 was identified as a caffeine-resistance gene by screening a genomic library of Saccharomyces cerevisiae in a multicopy vector YEp24. SNQ2 encodes an ATP-binding cassette transporter and is highly homologous to PDR5. Multicopy of PDR5 also conferred resistance to caffeine, while its resistance was smaller than that of SNQ2. Residual caffeine contents were analyzed after transiently exposing cells to caffeine. The ratios of caffeine contents were 21.3 ± 8.8% (YEp24-SNQ2) and 81.9 ± 8.7% (YEp24-PDR5) relative to control (YEp24, 100%). In addition, multicopies of SNQ2 or PDR5 conferred resistance to rhodamine 6G (R6G), which was widely used as a substrate for transport assay. R6G was exported by both transporters, and their efflux activities were inhibited by caffeine with half-maximal inhibitory concentrations of 5.3 ± 1.9 (YEp24-SNQ2) and 17.2 ± 9.6 mM (YEp24-PDR5). These results demonstrate that Snq2p is a more functional transporter of caffeine than Pdr5p in yeast cells.

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Section: Discussionmentioning

confidence: 99%

Multidrug resistance transporters Snq2p and Pdr5p mediate caffeine efflux in Saccharomyces cerevisiae

Tsujimoto

Shimizu

Otake

et al. 2015

Bioscience, Biotechnology, and Biochemistry

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“…The deviant Walker A substitution C199A was phenotypically WT. These results strongly suggested that the deviant ATP site is non-catalytic, an observation clearly supported by further work with Pdr5 [23,24]. The big surprise, however, was the behaviour of the alanine substitution in the highly conserved His 1068 of the H-loop.…”

Section: Important Features Of the Pdr5 Transportermentioning

confidence: 64%

“…Why such a substitution would appear in the N242K suppressor hunt was a mystery. However, subsequent work demonstrated that the atypical ATP site is heavily involved in signal transmission [23,24]. …”

Section: The Signal Interface Of Pdr5 Shares Some Features With Its Smentioning

confidence: 99%

“…The Tap1/Tap2 transporter appears to be an example of this phenomenon [53]. Alternatively, if substitution of canonical residues for atypical ones causes significant loss of function, which is certainly the case with Pdr5 [24], the atypical ATP-binding site is actually deviant.…”

Section: The Function Of Atypical Atp-binding Sites In Asymmetric Abcmentioning

confidence: 99%

“…Pdr5 research has made major original contributions to the study of ABC transporters, including the basis of substrate specificity [15–17], the path of the transmission interface between the ATP-and drug-binding sites [18–21], the biochemical role of deviant ATP-binding sites [22–24], and the probable gating or diode function [25]. In the present review, we focus on these important contributions.…”

Section: Introduction: Discovery Of Pdr5mentioning

confidence: 99%

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The multidrug transporter Pdr5 on the 25th anniversary of its discovery: an important model for the study of asymmetric ABC transporters

Golin

Ambudkar

2015

Biochemical Journal

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Asymmetric ABC (ATP-binding cassette) transporters make up a significant proportion of this important superfamily of integral membrane proteins. These proteins contain one canonical (catalytic) ATP-binding site and a second atypical site with little enzymatic capability. The baker’s yeast (Saccharomyces cerevisiae) Pdr5 multidrug transporter is the founding member of the Pdr subfamily of asymmetric ABC transporters, which exist only in fungi and slime moulds. Because these organisms are of considerable medical and agricultural significance, Pdr5 has been studied extensively, as has its medically important homologue Cdr1 from Candida albicans. Genetic and biochemical analyses of Pdr5 have contributed important observations that are likely to be applicable to mammalian asymmetric ABC multidrug transporter proteins, including the basis of transporter promiscuity, the function of the non-catalytic deviant ATP-binding site, the most complete description of an in vivo transmission interface, and the recent discovery that Pdr5 is a molecular diode (one-way gate). In the present review, we discuss the observations made with Pdr5 and compare them with findings from clinically important asymmetric ABC transporters, such as CFTR (cystic fibrosis transmembrane conductance regulator), Cdr1 and Tap1/Tap2.

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The role of the degenerate nucleotide binding site in type I ABC exporters

2020

Self Cite

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ATP-binding cassette (ABC) transporters are fascinating molecular machines that are capable of transporting a large variety of chemically diverse compounds. The energy required for translocation is derived from binding and hydrolysis of ATP. All ABC transporters share a basic architecture and are composed of two transmembrane domains and two nucleotide binding domains (NBDs). The latter harbor all conserved sequence motifs that hallmark the ABC transporter superfamily. The NBDs form the nucleotide binding sites (NBSs) in their interface. Transporters with two active NBSs are called canonical transporters, while ABC exporters from eukaryotic organisms, including humans, frequently have a degenerate NBS1 containing noncanonical residues that strongly impair ATP hydrolysis. Here, we summarize current knowledge on degenerate ABC transporters. By integrating structural information with biophysical and biochemical evidence of asymmetric function, we develop a model for the transport cycle of degenerate ABC transporters. We will elaborate on the unclear functional advantages of a degenerate NBS.

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Generating Symmetry in the Asymmetric ATP-binding Cassette (ABC) Transporter Pdr5 from Saccharomyces cerevisiae

Cited by 34 publications

References 35 publications

Multidrug resistance transporters Snq2p and Pdr5p mediate caffeine efflux in Saccharomyces cerevisiae

Multidrug resistance transporters Snq2p and Pdr5p mediate caffeine efflux in Saccharomyces cerevisiae

The multidrug transporter Pdr5 on the 25th anniversary of its discovery: an important model for the study of asymmetric ABC transporters

The role of the degenerate nucleotide binding site in type I ABC exporters

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