The Yeast ATP-binding Cassette (ABC) Transporter Ycf1p Enhances the Recruitment of the Soluble SNARE Vam7p to Vacuoles for Efficient Membrane Fusion

Sasser, Terry; Lawrence, Gus; Karunakaran, Surya; Brown, Christopher C.; Fratti, Rutilio A.

doi:10.1074/jbc.m112.441089

Cited by 20 publications

(19 citation statements)

References 56 publications

(70 reference statements)

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“…However, we found that the class C ABC transporter Ycf1 was also needed for Vam7 recruitment. The deletion of YCF1 led to a reduction in vacuolar Vam7 that was linked to inhibited fusion (30). Importantly, the fusion defect observed with ycf1⌬ vacuoles was rescued by …”

Section: The Polybasic Region Of the Vam7 Middle Domain Ismentioning

confidence: 87%

“…Vacuoles deleted in YCF1 were isolated from RFY32 and RFY33 (30). In vitro content mixing fusion reactions (30 l) contained 3 g each of vacuoles from BJ3505 (PHO8 pep4⌬) and DKY6281 (pho8⌬ PEP4) backgrounds, fusion reaction buffer (20 mM PIPES-KOH, pH 6.8, 200 mM sorbitol, 125 mM KCl, 5 mM MgCl 2 ), ATP regenerating system (1 mM ATP, 0.1 mg/ml creatine kinase, 29 mM creatine phosphate), 10 M coenzyme A, and 283 nM Pbi2.…”

Section: Methodsmentioning

confidence: 99%

“…Vam7-triggered fusion is blocked by the Q283R mutation in the ionic zero layer of the SNARE motif, which stalls the fusion pathway in a hemifusion state (18,29). Vam7 is also recruited to vacuoles through a mechanism dependent on the ABC transporter Ycf1 through an unknown mechanism (30). Vam7 contains a putative ␣ helical middle domain that is uncharacterized.…”

mentioning

confidence: 99%

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The Central Polybasic Region of the Soluble SNARE (Soluble N-Ethylmaleimide-sensitive Factor Attachment Protein Receptor) Vam7 Affects Binding to Phosphatidylinositol 3-Phosphate by the PX (Phox Homology) Domain

Miner

Starr²,

Hurst

et al. 2016

Journal of Biological Chemistry

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Section: The Polybasic Region Of the Vam7 Middle Domain Ismentioning

confidence: 87%

Section: Methodsmentioning

confidence: 99%

mentioning

confidence: 99%

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The Central Polybasic Region of the Soluble SNARE (Soluble N-Ethylmaleimide-sensitive Factor Attachment Protein Receptor) Vam7 Affects Binding to Phosphatidylinositol 3-Phosphate by the PX (Phox Homology) Domain

Miner

Starr²,

Hurst

et al. 2016

Journal of Biological Chemistry

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“…82 Deletion of Ycf1 caused a significant reduction in fusion efficiency that was linked to the exclusion of Vam7 from isolated ycf1D vacuoles, whereas other SNAREs, HOPS subunits and Ypt7 were uneffected. 83 The attenuated fusion was restored to wild type levels by the addition of recombinant Vam7, indicating that it was able to readily interact with other SNAREs, HOPS and lipids on the membrane to stimulate fusion. Because Vam7 associates with PI3P and Ycf1 might effect the PI3P levels during the fusion reaction we next asked whether the effect of deleting YCF1 on Vam7 recruitment was due to altered PI3P levels.…”

Section: Ycf1 and Fusionmentioning

confidence: 98%

Class C ABC transporters andSaccharomyces cerevisiaevacuole fusion

Sasser¹,

Fratti²

2014

Cellular Logistics

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“…S2 in the supplemental material). S. cerevisiae Ycf1p is an ABC transporter that transports a broad range of toxins into vacuoles (33,34). Transcription of A. fumigates abcB can be induced by voriconazole, and the abcB-knockout mutant is hypersensitive to 6 conidia/ml) of the N. crassa WT strain, the ads-4-knockout mutant (⌬ads-4), the ads-4-complemented strain (⌬ads-4::ads-4), and the ads-4 overexpression strain (ads-4 OE ) were spotted onto the center of Vogel's plates (diameter, 9 cm) with or without antifungal drugs.…”

Section: Ads-4 Responds Transcriptionally To Ketoconazole Inmentioning

confidence: 99%

Transcription Factor ADS-4 Regulates Adaptive Responses and Resistance to Antifungal Azole Stress

Wang

Zhang

Chen

et al. 2015

Antimicrob Agents Chemother

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bAzoles are commonly used as antifungal drugs or pesticides to control fungal infections in medicine and agriculture. Fungi adapt to azole stress by rapidly activating the transcription of a number of genes, and transcriptional increases in some azole-responsive genes can elevate azole resistance. The regulatory mechanisms that control transcriptional responses to azole stress in filamentous fungi are not well understood. This study identified a bZIP transcription factor, ADS-4 (antifungal drug sensitive-4), as a new regulator of adaptive responses and resistance to antifungal azoles. Transcription of ads-4 in Neurospora crassa cells increased when they were subjected to ketoconazole treatment, whereas the deletion of ads-4 resulted in hypersensitivity to ketoconazole and fluconazole. In contrast, the overexpression of ads-4 increased resistance to fluconazole and ketoconazole in N. crassa. Transcriptome sequencing (RNA-seq) analysis, followed by quantitative reverse transcription (qRT)-PCR confirmation, showed that ADS-4 positively regulated the transcriptional responses of at least six genes to ketoconazole stress in N. crassa. The gene products of four ADS-4-regulated genes are known contributors to azole resistance, including the major efflux pump CDR4 (Pdr5p ortholog), an ABC multidrug transporter (NcAbcB), sterol C-22 desaturase (ERG5), and a lipid transporter (NcRTA2) that is involved in calcineurin-mediated azole resistance. Deletion of the ads-4-homologous gene Afads-4 in Aspergillus fumigatus caused hypersensitivity to itraconazole and ketoconazole, which suggested that ADS-4 is a functionally conserved regulator of adaptive responses to azoles. This study provides important information on a new azole resistance factor that could be targeted by a new range of antifungal pesticides and drugs. F ilamentous fungi cause over 70% of plant diseases, and some can cause deadly infections in humans (1-5). Azoles (e.g., itraconazole [ITA], fluconazole [FLC], and ketoconazole [KTC]) are the most commonly used antifungal drugs in medicine, and some azoles, such as triadimenol and propiconazole, are also used to control fungal diseases in plants (6). Antifungal azoles inhibit 14␣-methyl sterol demethylase (encoded by ERG11), a key enzyme involved in fungal ergosterol biosynthesis. This leads to changes in membrane consistency (7,8). In addition to blocking ergosterol production, the inhibition of ERG11 by azoles results in the accumulation of toxic 14␣-methylated sterol intermediates (9, 10).Fungi can adapt to azole stress by rapidly increasing the expression of a number of genes. Increased expression of some azoleresponsive genes, such as genes encoding azole efflux pumps and genes involved in ergosterol biosynthesis, can increase resistance to azoles (11,12). Previous studies in Saccharomyces cerevisiae and Candida albicans identified a number of regulatory genes that mediate azole responses. The transcription factors Pdr1p and Pdr3p in S. cerevisiae and their homologs in C. albicans regulate azole responses by c...

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The Yeast ATP-binding Cassette (ABC) Transporter Ycf1p Enhances the Recruitment of the Soluble SNARE Vam7p to Vacuoles for Efficient Membrane Fusion

Cited by 20 publications

References 56 publications

The Central Polybasic Region of the Soluble SNARE (Soluble N-Ethylmaleimide-sensitive Factor Attachment Protein Receptor) Vam7 Affects Binding to Phosphatidylinositol 3-Phosphate by the PX (Phox Homology) Domain

The Central Polybasic Region of the Soluble SNARE (Soluble N-Ethylmaleimide-sensitive Factor Attachment Protein Receptor) Vam7 Affects Binding to Phosphatidylinositol 3-Phosphate by the PX (Phox Homology) Domain

Class C ABC transporters andSaccharomyces cerevisiaevacuole fusion

Transcription Factor ADS-4 Regulates Adaptive Responses and Resistance to Antifungal Azole Stress

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