Review: Subcellular traffic of the plasma membrane H+-ATPase in Saccharomyces cerevisiae

d’Exaerde, Alban de Kerchove; Supply, Philip; Goffeau, André

doi:10.1002/(sici)1097-0061(199608)12:10<907::aid-yea10>3.0.co;2-2

Cited by 29 publications

(20 citation statements)

References 59 publications

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“…Traditional overexpression of plasma membrane proteins using strong promoters and multicopy vectors often causes mistargeting and stimulates accumulation of intracellular membranes (30). These problems have been overcome in a new approach, which has allowed dramatically enhanced overexpression of Pdr5p and Snq2p ABC transporters in the yeast plasma membrane (6 -8).…”

Section: Resultsmentioning

confidence: 99%

ATPase and Multidrug Transport Activities of the Overexpressed Yeast ABC Protein Yor1p

Decottignies¹,

Grant²,

Nichols³

et al. 1998

Journal of Biological Chemistry

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The Saccharomyces cerevisiae genome encodes 15 fullsize ATP binding cassette transporters (ABC), of which PDR5, SNQ2, and YOR1 are known to be regulated by the transcription factors Pdr1p and Pdr3p (pleiotropic drug resistance). We have identified two new ABC transporter-encoding genes, PDR10 and PDR15, which were upregulated by the PDR1-3 mutation. These genes, as well as four other ABC transporter-encoding genes, were deleted in order to study the properties of Yor1p. The PDR1-3 gain-of-function mutant was then used to overproduce Yor1p up to 10% of the total plasma membrane proteins. Despite their different topologies, both Yor1p and Pdr5p mediated the ATP-dependent translocation of similar drugs and phospholipids across the yeast cell membrane. Both ABC transporters exhibit ATP hydrolysis in vitro, but Pdr5p ATPase activity is about 15 times higher than that of Yor1p, which may indicate mechanistic or regulatory differences between the two enzymes. The yeast YOR11 gene confers oligomycin resistance on overexpression in a 2-m plasmid (1). Its nucleotide sequence reveals an ORF of 1477 amino acids encoding an ABC protein highly homologous to mammalian transporters such as the multidrug resistance-conferring enzyme MRP (BLAST (see Ref. 2) sequence homology score: p ϭ e Ϫ228 ), the organic anion transporter cMOAT (p ϭ e Ϫ216 ), the sulfonylurea receptor (p ϭ e Ϫ164 ), and the cystic fibrosis transmembrane conductance regulator CFTR (p ϭ e Ϫ132 ). Yor1p is a "full-size" ABC transporter with the topology (TM-NBF) 2 (3, 4). It consists of two homologous halves, with each containing a putative ATP-binding domain (NBF) and a transmembrane domain of six membrane spans (TM). Cui et al. (5) showed that Yor1p confers resistance to a series of drugs including reveromycin A and suggested that Yor1p may be involved in the cellular efflux of organic anions including the fluorescent dye rhodamine B. They also showed that incubation with reveromycin A increases the YOR1 mRNA level. The transcription of YOR1 is controlled by the homologous pair of transcription factors Pdr1p/Pdr3p. The level of YOR1 transcription is decreased by the deletion of either PDR1 or PDR3 and increased in the presence of the gain-of-function PDR1 alleles (1).In this paper, we have investigated the transport activity of Yor1p. Building on previous studies, which indicated that the (TM-NBF) 2 -type Yor1p, together with the (NBF-TM) 2 -type Pdr5p and Snq2p ABC transporters, are overexpressed in the PDR1-3 mutant plasma membrane (6 -8), the PDR1-3 mutant has been used as a tool that enhances the Yor1p protein level. As another investigative tool, we constructed a set of isogenic strains, in the PDR1-3 mutant, with multiple deletions of homologous ABC genes since, in situations where two or more proteins located in the same subcellular compartment share a common substrate, a clear phenotype is only seen when all the corresponding genes are deleted, as illustrated by the work of Mahé et al. (9), who showed that Pdr5p and Snq2p have an overlapping transport ...

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Section: Resultsmentioning

confidence: 99%

ATPase and Multidrug Transport Activities of the Overexpressed Yeast ABC Protein Yor1p

Decottignies¹,

Grant²,

Nichols³

et al. 1998

Journal of Biological Chemistry

Self Cite

360

334

View full text Add to dashboard Cite

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“…In the mutant D378A and other Asp substitutions, the targeting of newly synthesized Pma1 to the plasma membrane is impaired (42)(43)(44). It was suggested that most of the mutant protein was misfolded and retained in the ER, due to its quality control in yeast (25,45). Another example of a reduced amount of Pma1p in plasma membrane and its retention in the ER is in the Lst1p mutant (27).…”

Section: Altered Distribution Of Yeast Plasma Membrane Hmentioning

confidence: 99%

“…One goes through the endosomes, and the other delivers the proteins directly to the vacuole from the post-Golgi vesicles. Similarly, multiple delivery pathways of plasma membrane proteins may exist (25). Studies of Pma1p sorting revealed involvement of several proteins needed specifically for its transport to the plasma membrane (9,26,27).…”

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confidence: 99%

Altered Distribution of the Yeast Plasma Membrane H+-ATPase as a Feature of Vacuolar H+-ATPase Null Mutants

Perzov

Nelson

2000

Journal of Biological Chemistry

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“…At first glance, this idea fits with the striking proliferation of ER membranes that takes place during overexpression of the closely related PMA2 ATPase (32), H ϩ -ATPases from higher plants (39), and the integral ER proteins hydroxymethylglutaryl-coenzyme A reductase (31) and cytochrome b 5 (40). As reviewed by Kerchove et al (41), the morphology of the proliferated ER can vary, depending on the expressed protein, but in all cases is thought to occur as the cell's response to increased levels of these proteins in the ER.…”

Section: Phosphorylation Region Of the Yeast Pma1 Hmentioning

confidence: 99%

“…This point was shown in a recent study in which secretory vesicles, isolated from cells producing the D378N ATPase, contained a normal profile of Coomassie-stained proteins (18), suggesting that the biosynthetic block is limited to the ATPase or a small subset of secretory proteins. Two explanations have been proposed for the seeming specificity of the block (16,41). First, the wild-type ATPase may form oligomers with malfolded mutant ATPase, resulting in the retention of both.…”

Section: Phosphorylation Region Of the Yeast Pma1 Hmentioning

confidence: 99%

Phosphorylation Region of the Yeast Plasma-membrane H+-ATPase

DeWitt

Santos

Allen

et al. 1998

Journal of Biological Chemistry

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Mutations at the phosphorylation site (Asp-378) of the yeast plasma-membrane H ؉ -ATPase have been shown previously to cause misfolding of the ATPase, preventing normal movement along the secretory pathway; Asp-378 mutations also block the biogenesis of co-expressed wild-type ATPase and lead to a dominant lethal phenotype. To ask whether these defects are specific for Asp-378 or whether the phosphorylation region as a whole is involved, alanine-scanning mutagenesis has been carried out to examine the role of 11 conserved residues flanking Asp-378. In the sec6 -4 expression system (Nakamoto, R. K., Rao, R., and Slayman, C. W. (1991) J. Biol. Chem. 266, 7940 -7949), the mutant ATPases displayed varying abilities to reach the secretory vesicles that deliver plasma-membrane proteins to the cell surface. Indirect immunofluorescence of intact cells also gave evidence for a spectrum of behavior, ranging from mutant ATPases completely arrested (D378A, K379A, T380A, and T384A) or partially arrested in the endoplasmic reticulum to those that reached the plasma membrane in normal amounts (C376A, S377A, and G381A). Although the extent of ER retention varied among the mutants, the endoplasmic reticulum appeared to be the only secretory compartment in which the mutant ATPases accumulated. All of the mutant proteins that localized either partially or fully to the ER were also malfolded based on their abnormal sensitivity to trypsin. Among them, the severely affected mutants had a dominant lethal phenotype, and even the intermediate mutants caused a visible slowing of growth when co-expressed with wild-type ATPase. The effects on growth could be traced to the trapping of the wild-type enzyme with the mutant enzyme in the ER, as visualized by double label immunofluorescence. Taken together, the results indicate that the residues surrounding Asp-378 are critically important for ATPase maturation and transport to the cell surface.

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Review: Subcellular traffic of the plasma membrane H+-ATPase in Saccharomyces cerevisiae

Cited by 29 publications

References 59 publications

ATPase and Multidrug Transport Activities of the Overexpressed Yeast ABC Protein Yor1p

ATPase and Multidrug Transport Activities of the Overexpressed Yeast ABC Protein Yor1p

Altered Distribution of the Yeast Plasma Membrane H+-ATPase as a Feature of Vacuolar H+-ATPase Null Mutants

Phosphorylation Region of the Yeast Plasma-membrane H+-ATPase

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