Isolation and characterization of a yeast gene, <i>MPD1</i>, the overexpression of which suppresses inviability caused by protein disulfide isomerase depletion

Tachikawa, Hiroyuki; Takeuchi, Yutaka; Funahashi, Wataru; Miura, Tadashi; Gao, Xiao‐Dong; Fujimoto, Daisaburo; Mizunaga, Takemitsu; Onodera, Kônoshin

doi:10.1016/0014-5793(95)00750-4

Cited by 58 publications

(40 citation statements)

References 38 publications

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“…However, we found that the substrate-binding domain (BЈ) was not required for productive interaction with Ero1p. We also found that Ero1p is fairly nonspecific in its recognition of thioredoxin domains, which helps explain why the expression of several PDI homologs can rescue the viability of a ⌬pdi1 strain (11,12,37). This lack of discrimination seems at odds with the strong preference that Ero1p has for thioredoxin domains over unfolded proteins (19).…”

Section: Discussionmentioning

confidence: 63%

Domain Architecture of Protein-disulfide Isomerase Facilitates Its Dual Role as an Oxidase and an Isomerase in Ero1p-mediated Disulfide Formation

Kulp

Frickel

Ellgaard

et al. 2006

Journal of Biological Chemistry

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Native disulfide bond formation in eukaryotes is dependent on protein-disulfide isomerase (PDI) and its homologs, which contain varying combinations of catalytically active and inactive thioredoxin domains. However, the specific contribution of PDI to the formation of new disulfides versus reduction/rearrangement of non-native disulfides is poorly understood. We analyzed the role of individual PDI domains in disulfide bond formation in a reaction driven by their natural oxidant, Ero1p. We found that Ero1p oxidizes the isolated PDI catalytic thioredoxin domains, A and A at the same rate. In contrast, we found that in the context of full-length PDI, there is an asymmetry in the rate of oxidation of the two active sites. This asymmetry is the result of a dual effect: an enhanced rate of oxidation of the second catalytic (A) domain and the substratemediated inhibition of oxidation of the first catalytic (A) domain. The specific order of thioredoxin domains in PDI is important in establishing the asymmetry in the rate of oxidation of the two active sites thus allowing A and A, two thioredoxin domains that are similar in sequence and structure, to serve opposing functional roles as a disulfide isomerase and disulfide oxidase, respectively. These findings reveal how native disulfide folding is accomplished in the endoplasmic reticulum and provide a context for understanding the proliferation of PDI homologs with combinatorial arrangements of thioredoxin domains.Proteins that traverse the secretory pathway typically contain disulfide bonds that are critical for their correct fold and function. In eukaryotes, the endoplasmic reticulum (ER) 2 is the entry point into the secretory pathway and is the cellular compartment where folding and disulfide bond formation occur (1, 2). Disulfides can form spontaneously in vitro in the presence of an oxidizing agent such as molecular oxygen or oxidized glutathione; however, this process is typically slow and inefficient. In vivo, disulfide bond formation is dependent on cellular machinery to catalyze the formation of new disulfides (oxidation) and the rearrangement of non-native disulfides (isomerization). Both oxidation and isomerization are necessary for allowing the full complement of native disulfide bond formation.Protein-disulfide isomerase (PDI), which was identified more than 40 years ago, plays a critical role in promoting native disulfide bond formation in vivo (3). PDI, an essential enzyme with the ability to catalyze both the oxidation of new disulfides and the isomerization of existing disulfides, is composed of four thioredoxin-like domains (4). The first and last domains (referred to as A and AЈ, respectively) contain Cys-x-x-Cys (CxxC) active sites, whereas the two middle domains (referred to as B and BЈ) are catalytically inactive (5, 6). Oxidation involves the transfer of an active site disulfide from PDI to substrate proteins, while isomerization requires the active site cysteines to be in a reduced form so that they can attack non-native disulfides in substrate ...

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Section: Discussionmentioning

confidence: 63%

Domain Architecture of Protein-disulfide Isomerase Facilitates Its Dual Role as an Oxidase and an Isomerase in Ero1p-mediated Disulfide Formation

Kulp

Frickel

Ellgaard

et al. 2006

Journal of Biological Chemistry

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“…The S. cerevisiae hut1 single disruptant is derived from W303-1A. FW14 (Tachikawa et al, 1995), FW24, ero1 ts mutant, and hut1/ero1 double mutant are derived from YPH500. The S. pombe cells were cultured in YPD, YES or MM media (Moreno et al, 1991) and S. cerevisiae cells were cultured in YPD, YPGal or SD media (Sherman, 1991).…”

Section: Methodsmentioning

confidence: 99%

“…Plasmids pmMPD1 and pmMPD2, in which MPD1 and MPD2 are cloned into pYO325, were constructed as described previously (Tachikawa et al, 1995(Tachikawa et al, , 1997.…”

Section: Plasmid Constructionmentioning

confidence: 99%

“…This indicates that Ero1p, Pdi1p and Figure 6 shows that scHUT1 has genetic interactions not only with ERO1 but also with PDI-related genes. In the S. cerevisiae strain FW14, chromosomal PDI1 is placed under the control of GAL1 promoter (Tachikawa et al, 1995). Further, the chromosomal HUT1 gene in FW14 was disrupted to construct FW24 strain (data not shown).…”

Section: Schut1 Has Genetic Interactions With Ero1 and Pdi-related Genesmentioning

confidence: 99%

“…Although the essential function of Pdi1p in yeast cells is considered to be the isomerization of non-native disulphide bonds (Laboissiere et al, 1995), several lines of evidence indicate that Pdi1p also catalyses the oxidation of secretory proteins (Lyles and Gilbert, 1991;Frand and Kaiser, 1999). S. cerevisiae carries four additional PDI-related proteins (Mpd1p, Mpd2p, Eug1p and Eps1p) (Tachikawa et al, 1995(Tachikawa et al, , 1997Tachibana and Stevens, 1992;Wang and Chang, 1999). Mpd1p, Mpd2p and Eps1p have a single active-site motif of CXXC, and Eug1p has two active-site motifs of CXXS.…”

Section: Introductionmentioning

confidence: 99%

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Hut1 proteins identified in Saccharomyces cerevisiae and Schizosaccharomyces pombe are functional homologues involved in the protein‐folding process at the endoplasmic reticulum

Nakanishi

Nakayama

Yokota

et al. 2001

Yeast

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The Saccharomyces cerevisiae HUT1 gene (scHUT1) and the Schizosaccharomyces pombe hut1+ gene (sphut1 + ) encode hydrophobic proteins with approximately 30% identity to a human UDP-galactose transporter-related gene (UGTrel1) product. These proteins show a significant similarity to the nucleotide sugar transporter and are conserved in many eukaryotic species, but their physiological functions are not known. Both scHUT1 and sphut1+ genes are non-essential for cell growth under normal conditions, and their disruptants show no defects in the modification of O-and N-linked oligosaccharides, but are sensitive to a membrane-permeable reducing agent, dithiothreitol (DTT). Consistent with this phenotype, scHUT1 has genetic interaction with ERO1, which plays an essential role in the oxidation of secretory proteins at the endoplasmic reticulum (ER). Overexpression of the MPD1 or MPD2 genes, which were isolated as multicopy suppressors of protein disulphide isomerase (PDI) depletion, could not replace the essential function of PDI in Dhut1 S. cerevisiae cells. Our results indicate that scHut1p and spHut1p are functional homologues, and their physiological function is to maintain the optimal environment for the folding of secretory pathway proteins in the ER.

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Protein disulphide isomerase genes ofKluyveromyces lactis

Bao

Huo

et al. 2000

Yeast

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Two genes of Kluyveromyces lactis, KlPDI1 and KlMPD1, were studied. They code for a protein disulphide isomerase and its structural and functional homologue, respectively. The KlPDI1 product was 52.6% identical to Pdi1p and the KlMPD1 product 47% identical to Mpd1p of S. cerevisiae. Both genes contained the thioredoxin‐active site‐related signature. Their C‐termini showed a new variant of the endoplasmic reticulum‐retention signal, QDEL. A single copy of KlPDI1 was able to complement the growth defect of a pdi1 mutation. KlMPD1 on a multicopy vector partially suppressed the klpdi1 and pdi1 mutations. The Klpdi1 null mutation was lethal. The klmpd1 disruptant was viable, but showed an increased sensitivity to high temperature. Several stress response motifs were present in the upstream sequence of KlMPD1, but not of KlPDI1, whilst the opposite is known for the S. cerevisiae homologues. The viability of the klmpd1 mutant under starvation for nitrogen or carbon source was not different from that of the wild‐type. The syntenic relationship is discussed for the KlPDI1 gene regions with respect to the duplicated segments PDI1/EUG1 in S. cerevisiae. EMBL Accession Nos: AJ243958 (KlPDI1 region) and AJ243960 (KlMPD1 region). Copyright © 2000 John Wiley & Sons, Ltd.

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Isolation and characterization of a yeast gene, MPD1, the overexpression of which suppresses inviability caused by protein disulfide isomerase depletion

Cited by 58 publications

References 38 publications

Domain Architecture of Protein-disulfide Isomerase Facilitates Its Dual Role as an Oxidase and an Isomerase in Ero1p-mediated Disulfide Formation

Domain Architecture of Protein-disulfide Isomerase Facilitates Its Dual Role as an Oxidase and an Isomerase in Ero1p-mediated Disulfide Formation

Hut1 proteins identified in Saccharomyces cerevisiae and Schizosaccharomyces pombe are functional homologues involved in the protein‐folding process at the endoplasmic reticulum

Protein disulphide isomerase genes ofKluyveromyces lactis

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