SUC1 and SUC2: two sucrose transporters from <i>Arabidopsis thaliana</i>; expression and characterization in baker's yeast and identification of the histidine‐tagged protein

Sauer, Norbert; Stolz, JLirgen

doi:10.1046/j.1365-313x.1994.6010067.x

Cited by 330 publications

(332 citation statements)

References 38 publications

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“…The resulting strain (referred to as the gap1 mutant) was maintained on yeast/peptone/dextrose medium containing 1% (w/v) yeast extract, 2% (w/v) peptone, 2% (w/v) dextrose and 2% (w/v) agar. RcAAP3 cDNA was excised with NotI (using the internal restriction sites incorporated into adapters during cDNA library construction) and cloned into the E. coli/yeast expression vector NEV-N (Sauer and Stolz, 1994). NEV-N/RcAAP3 clones containing cDNAs in sense and antisense orientations in relation to the constitutive PMA1 promoter in the vector were identified by restriction analysis.…”

Section: Functional Complementation In a Yeast Transport Mutantmentioning

confidence: 99%

Functional Characterization and Expression Analysis of the Amino Acid Permease RcAAP3 from Castor Bean1

Neelam¹,

Marvier

Hall

et al. 1999

Plant Physiology

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A polymerase chain reaction-based library screening procedure was used to isolate RcAAP3, an amino acid permease cDNA from castor bean (Ricinus communis). RcAAP3 is 1.7 kb in length, with an open reading frame that encodes a protein with a calculated molecular mass of 51 kD. Hydropathy analysis indicates that the RcAAP3 protein is highly hydrophobic in nature with nine to 11 putative transmembrane domains. RcAAP3-mediated uptake of citrulline in a yeast transport mutant showed saturable kinetics with a K m of 0.4 mM. Transport was higher at acidic pH and was inhibited by the protonophore carbonylcyanide-m-chlorophenylhydrazone, suggesting a proton-coupled transport mechanism. Citrulline uptake was strongly inhibited (72%) by the permeable sulfydryl reagent N-ethylmaleimide, but showed lower sensitivity (30% inhibition) to the nonpermeable reagent p-chloromercuribenzenesulfonic acid. Diethylpyrocarbonate, a histidine modifier, inhibited citrulline uptake by 80%. A range of amino acids inhibited citrulline uptake, suggesting that RcAAP3 may be a broad substrate permease that can transport neutral and basic amino acids with a lower affinity for acidic amino acids. Northern analysis indicated that RcAAP3 is widely expressed in source and sink tissues of castor bean, and that the pattern of expression is distinct from RcAAP1 and RcAAP2.

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Section: Functional Complementation In a Yeast Transport Mutantmentioning

confidence: 99%

Functional Characterization and Expression Analysis of the Amino Acid Permease RcAAP3 from Castor Bean1

Neelam¹,

Marvier

Hall

et al. 1999

Plant Physiology

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“…These models can be combined into a single model in which the binding on the external side can be random, but it can only be ordered on the inside, with the sugar dissociating before the proton. When SUC1 was expressed in yeast, it was found to be equally active at pH 5 or 6 and the relative activity decreased to 50% when the pH was increased to 7 (Sauer & Stolz, 1994), but these values do not take account of changes in V m , which in yeast is likely to vary with external pH. Furthermore, for SUC1 in yeast the value of K S m was 0.45 mM at an external pH of 5.5 (Sauer & Stolz, 1994), which is remarkably similar to the values obtained for the carrier expressed in oocytes.…”

Section: General Commentsmentioning

confidence: 99%

“…When SUC1 was expressed in yeast, it was found to be equally active at pH 5 or 6 and the relative activity decreased to 50% when the pH was increased to 7 (Sauer & Stolz, 1994), but these values do not take account of changes in V m , which in yeast is likely to vary with external pH. Furthermore, for SUC1 in yeast the value of K S m was 0.45 mM at an external pH of 5.5 (Sauer & Stolz, 1994), which is remarkably similar to the values obtained for the carrier expressed in oocytes. The stoichiometry of the H + / sucrose symport appears to be 1:1 and this agrees with measurements made using plasma membrane vesicles (e.g., Bush, 1990), but there are also reports of a variable stochiometry (e.g., Komor, 1997).…”

Section: General Commentsmentioning

confidence: 99%

“…The strain was engineered to express a cytoplasmic form of yeast invertase, which then allowed cells which were expressing a sucrose carrier to grow when the only carbon source was sucrose. The spinach cDNA was then used to probe an Arabidopsis thaliana library and to isolate two related sucrose transporters, SUC1 and SUC2 (Sauer & Stolz, 1994). Yeast cells transformed with SUC1 were able to accumulate the 14 C-labeled sucrose, whereas cells transformed with the gene in antisense orientation were not (Sauer & Stolz, 1994).…”

Section: Introductionmentioning

confidence: 99%

“…The spinach cDNA was then used to probe an Arabidopsis thaliana library and to isolate two related sucrose transporters, SUC1 and SUC2 (Sauer & Stolz, 1994). Yeast cells transformed with SUC1 were able to accumulate the 14 C-labeled sucrose, whereas cells transformed with the gene in antisense orientation were not (Sauer & Stolz, 1994). Furthermore, this uptake was shown to be pH-dependent with uptake increasing with proton concentration but saturating between pH 5 and 6.…”

Section: Introductionmentioning

confidence: 99%

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A Kinetic Model with Ordered Cytoplasmic Dissociation for SUC1, an Arabidopsis H + /Sucrose Cotransporter Expressed in Xenopus Oocytes

Zhou

Theodoulou

Sauer

et al. 1997

Journal of Membrane Biology

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Abstract.To elucidate the kinetic properties of the Arabidopsis H + /sucrose cotransporter, SUC1, with respect to transmembrane voltage and ligand concentrations, the transport system was heterologously expressed in Xenopus laevis oocytes. decreased as a function of the external H + concentration. Eight six-state carrier models-which comprised the four possible permutations of intracellular and extracellular ligand binding order, each with charge translocation on the sugar-loaded or -unloaded forms of the carrier-were analyzed algebraically with respect to their competence to account for the ensemble of kinetic observations. Of these, two models (first-on, first-off and last-on, first-off with respect to sucrose binding as it passes from outside to inside the cell and with charge translocation on the loaded form of the carrier) exhibit sufficient kinetic flexibility to describe the observations. Combining these two, a single model emerges in which the binding on the external side can be random, but it can only be ordered on the inside, with the sugar dissociating before the proton.

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Post-translational fate ofCAN1 permease ofSaccharomyces cerevisiae

Opekarová

Caspari

Pinson

et al. 1998

Yeast

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To study the post‐translational fate of arginine permease (Can1p), the gene coding for this transport protein was placed behind a constitutive promoter of plasma membrane ATPase (PMA1) and furnished with a Myc tag. In exponential‐phase cells the amount of Can1p is constant, although turnover can be demonstrated. A rapid decrease in transport activity during the early stationary phase is paralleled by a corresponding net degradation of the protein. The amount of Can1p present in exponential cells grown on various nitrogen sources is the same, except in arginine‐grown cells, in which the amount of the protein is markedly lower. This occurs solely when arginine serves as nitrogen source but not as an immediate consequence of, for example, arginine addition to cells growing on other nitrogen sources. It was demonstrated that Can1p is phosphorylated. Since Can1p expression under the PMA1 promoter is glucose‐dependent, the amount of the permease expressed in high‐glucose‐grown cells is higher than in low‐glucose‐grown ones. Only a part of the Can1p overexpressed in high‐glucose‐grown cells is phosphorylated, while in low‐glucose‐grown cells the phosphorylated form probably represents the majority of Can1p. The permease phosphorylation or dephosphorylation is not related to transinhibition. © 1998 John Wiley & Sons, Ltd.

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SUC1 and SUC2: two sucrose transporters from Arabidopsis thaliana; expression and characterization in baker's yeast and identification of the histidine‐tagged protein

Cited by 330 publications

References 38 publications

Functional Characterization and Expression Analysis of the Amino Acid Permease RcAAP3 from Castor Bean1

Functional Characterization and Expression Analysis of the Amino Acid Permease RcAAP3 from Castor Bean1

A Kinetic Model with Ordered Cytoplasmic Dissociation for SUC1, an Arabidopsis H + /Sucrose Cotransporter Expressed in Xenopus Oocytes

Post-translational fate ofCAN1 permease ofSaccharomyces cerevisiae

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