Differential regulation of two <i>myo</i>‐inositol transporter genes of <i>Saccharomyces cerevisiae</i>

Nikawa, Jùn‐ichi; Hosaka, K.; Yamashita, Shinji

doi:10.1111/j.1365-2958.1993.tb00967.x

Cited by 85 publications

(98 citation statements)

References 28 publications

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“…In a first approach we tried to determine the functional properties of AtINT2 in the bakers' yeast mutant D458-1B (Nikawa et al, 1991). This strain carries mutations in the ITR1 (inositol transporter) and INO1 (inositol-1-P synthase) genes and can grow only on high extracellular concentrations of myoinositol.…”

Section: Expression Of the Atint2 Cdna In Yeastmentioning

confidence: 99%

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Arabidopsis INOSITOL TRANSPORTER2 Mediates H⁺ Symport of Different Inositol Epimers and Derivatives across the Plasma Membrane

et al. 2007

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Of the four genes of the Arabidopsis (Arabidopsis thaliana) INOSITOL TRANSPORTER family (AtINT family) so far only AtINT4 has been described. Here we present the characterization of AtINT2 and AtINT3. cDNA sequencing revealed that the AtINT3 gene is incorrectly spliced and encodes a truncated protein of only 182 amino acids with four transmembrane helices. In contrast, AtINT2 codes for a functional transporter. AtINT2 localization in the plasma membrane was demonstrated by transient expression of an AtINT2-GREEN FLUORESCENT PROTEIN fusion in Arabidopsis and tobacco (Nicotiana tabacum) epidermis cells and in Arabidopsis protoplasts. Its functional and kinetic properties were determined by expression in yeast (Saccharomyces cerevisiae) cells and Xenopus laevis oocytes. Expression of AtINT2 in a Ditr1 (inositol uptake)/Dino1 (inositol biosynthesis) double mutant of bakers' yeast complemented the deficiency of this mutant to grow on low concentrations of myoinositol. In oocytes, AtINT2 mediated the symport of H 1 and several inositol epimers, such as myoinositol, scylloinositol, D-chiroinositol, and mucoinositol. The preference for individual epimers differed from that found for AtINT4. Moreover, AtINT2 has a lower affinity for myoinositol (K m 5 0.7-1.0 mM) than AtINT4 (K m 5 0.24 mM), and the K m is slightly voltage dependent, which was not observed for AtINT4. Organ and tissue specificity of AtINT2 expression was analyzed in AtINT2 promoter/reporter gene plants and showed weak expression in the anther tapetum, the vasculature, and the leaf mesophyll. A T-DNA insertion line (Atint2.1) and an Atint2.1/Atint4.2 double mutant were analyzed under different growth conditions. The physiological roles of AtINT2 are discussed.

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Section: Expression Of the Atint2 Cdna In Yeastmentioning

confidence: 99%

“…For expression of AtINT2 cDNAs in yeast (Saccharomyces cerevisiae) we used strains D458-1B (Nikawa et al, 1991) or SEY2102 (Emr et al, 1983). Escherichia coli DH5a (Hanahan, 1983) was used for all cloning steps, fusion proteins for antisera were made in E. coli Rosetta (Novagen).…”

Section: Strains and Growth Conditionsmentioning

confidence: 99%

Arabidopsis INOSITOL TRANSPORTER2 Mediates H⁺ Symport of Different Inositol Epimers and Derivatives across the Plasma Membrane

et al. 2007

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“…Inositol uptake from the medium is actively catalysed by two transporters, Itr1p and Itr2p. 192 Transcription of the ITR1 gene responds to regulation by inositol. 131,191 When inositol is present in the medium the activity of Itr1p is reduced.…”

Section: Phospholipid Synthesismentioning

confidence: 99%

“…192 Transcription of the ITR1 gene responds to regulation by inositol. 131,191 When inositol is present in the medium the activity of Itr1p is reduced. The transcriptional regulation of ITR1 depends on the INO2, INO4 and OPI1 genes.…”

Section: Phospholipid Synthesismentioning

confidence: 99%

Biochemistry, cell biology and molecular biology of lipids ofSaccharomyces cerevisiae

Daum

Lees

Bard

et al. 1998

Yeast

579

380

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The yeast Saccharomyces cerevisiae is a powerful experimental system to study biochemical, cell biological and molecular biological aspects of lipid synthesis. Most but not all genes encoding enzymes involved in fatty acid, phospholipid, sterol or sphingolipid biosynthesis of this unicellular eukaryote have been cloned, and many gene products have been functionally characterized. Less information is available about genes and gene products governing the transport of lipids between organelles and within membranes, turnover and degradation of complex lipids, regulation of lipid biosynthesis, and linkage of lipid metabolism to other cellular processes. Here we summarize current knowledge about lipid biosynthetic pathways in S. cerevisiae and describe the characteristic features of the gene products involved. We focus on recent discoveries in these fields and address questions on the regulation of lipid synthesis, subcellular localization of lipid biosynthetic steps, cross‐talk between organelles during lipid synthesis and subcellular distribution of lipids. Finally, we discuss distinct functions of certain key lipids and their possible roles in cellular processes. © 1998 John Wiley & Sons, Ltd.

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“…44,45 The other major route of inositol supply in yeast is uptake from the medium by the principal inositol transporter Itr1p. 46 Phosphatidylinositol is a direct precursor of phosphoinositolcontaining sphingolipids, which are synthesized on the luminal aspect of the Golgi, 47 and sphingolipid biosynthesis is controlled, at least in part, by the supply of phosphatidylinositol. 48 As Drs2p corresponds to a prominent aminophospholipid transport activity in yeast, 13,14 its association with both Ino1p and Itr1p may be relevant for the establishment of a proper transbilayer lipid arrangement in membranes that pass through the Golgi complex on their way to the plasma membrane.…”

Section: Discussionmentioning

confidence: 99%

A P₄-ATPase Protein Interaction Network Reveals a Link between Aminophospholipid Transport and Phosphoinositide Metabolism

et al. 2009

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High-throughput analysis of protein-protein interactions can provide unprecedented insight into how cellular processes are integrated at the molecular level. Yet membrane proteins are often overlooked in these studies owing to their hydrophobic nature and low abundance. Here we used a proteomicsbased strategy with the specific intention of identifying membrane-associated protein complexes. One important aspect of our approach is the use of chemical cross-linking to capture transient and lowaffinity protein interactions that occur in living cells prior to cell lysis. We applied this method to identify binding partners of the yeast Golgi P 4 -ATPase Drs2p, a member of a conserved family of putative aminophospholipid transporters. Drs2p was endogeneously tagged with both a polyhistidine and a biotinylation peptide, allowing tandem-affinity purification of Drs2p-containing protein complexes under highly stringent conditions. Mass-spectrometric analysis of isolated complexes yielded one known and nine novel Drs2p binding partners. Binding specificity was verified by an orthogonal in vivo membrane protein interaction assay, confirming the efficacy of our method. Strikingly, three of the novel Drs2p interactors are involved in phosphoinositide metabolism. One of these, the phosphatidylinositol-4-phosphatase Sac1p, also displays genetic interactions with Drs2p. Together, these findings suggest that aminophospholipid transport and phosphoinositide metabolism are interconnected at the Golgi.

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Differential regulation of two myo‐inositol transporter genes of Saccharomyces cerevisiae

Cited by 85 publications

References 28 publications

Arabidopsis INOSITOL TRANSPORTER2 Mediates H⁺ Symport of Different Inositol Epimers and Derivatives across the Plasma Membrane

Arabidopsis INOSITOL TRANSPORTER2 Mediates H⁺ Symport of Different Inositol Epimers and Derivatives across the Plasma Membrane

Biochemistry, cell biology and molecular biology of lipids ofSaccharomyces cerevisiae

A P₄-ATPase Protein Interaction Network Reveals a Link between Aminophospholipid Transport and Phosphoinositide Metabolism

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Differential regulation of two myo‐inositol transporter genes of Saccharomyces cerevisiae

Cited by 85 publications

References 28 publications

Arabidopsis INOSITOL TRANSPORTER2 Mediates H+ Symport of Different Inositol Epimers and Derivatives across the Plasma Membrane

Arabidopsis INOSITOL TRANSPORTER2 Mediates H+ Symport of Different Inositol Epimers and Derivatives across the Plasma Membrane

Biochemistry, cell biology and molecular biology of lipids ofSaccharomyces cerevisiae

A P4-ATPase Protein Interaction Network Reveals a Link between Aminophospholipid Transport and Phosphoinositide Metabolism

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Arabidopsis INOSITOL TRANSPORTER2 Mediates H⁺ Symport of Different Inositol Epimers and Derivatives across the Plasma Membrane

Arabidopsis INOSITOL TRANSPORTER2 Mediates H⁺ Symport of Different Inositol Epimers and Derivatives across the Plasma Membrane

A P₄-ATPase Protein Interaction Network Reveals a Link between Aminophospholipid Transport and Phosphoinositide Metabolism