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Cloning, characterization and regulation of a protein disulfide isomerase from the fission yeast Schizosaccharomyces pombe

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Abstract

To elucidate the physiological roles and regulation of a protein disulfide isomerase (PDI) from the fission yeast Schizosaccharomyces pombe, the full-length PDI gene was ligated into the shuttle vector pRS316, resulting in pPDI10. The determined DNA sequence carries 1,636 bp and encodes the putative 359 amino acid sequence of PDI with a molecular mass of 39,490 Da. In the amino acid sequence, the S. pombe PDI appears to be very homologous to A. thaliana PDI. The S. pombe cells harboring pPDI10 showed increased PDI activity and accelerated growth, suggesting that the cloned PDI gene is functioning and involved in the yeast growth. The 460 bp upstream region of the PDI gene was fused into promoterless β-galactosidase gene of the shuttle vector YEp367R to generate pYUPDI10. The synthesis of β-galactosidase from the PDI–lacZ fusion gene was enhanced by oxidative stress, such as superoxide anion and hydrogen peroxide. It was also induced by some non-fermentable and fermentable carbon sources. Nitrogen starvation was able to enhance the synthesis of β-galactosidase from the PDI–lacZ fusion gene. The enhancement by oxidative stress and fermentable carbon sources did not depend on the presence of Pap1. The PDI mRNA levels were increased in both Pap1-positive and Pap1-negative cells treated with glycerol. Taken together, the S. pombe PDI gene is involved in cellular growth and response to nutritional and oxidative stress.

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Abbreviations

ONPG:

o-nitrophenyl-β-d-galactoside

PCR:

polymerase chain reaction

PDI:

protein disulfide isomerase

RT-PCR:

reverse transcriptase-polymerase chain reaction

S. pombe :

Schizosaccharomyces pombe

References

  1. HF Gilbert (1997) ArticleTitleProtein disulfide isomerase and assisted protein folding J Biol Chem 272 29399–29402 Occurrence Handle9367991 Occurrence Handle1:CAS:528:DyaK2sXnsFWgtLw%3D Occurrence Handle10.1074/jbc.272.47.29399

    Article  PubMed  CAS  Google Scholar 

  2. PS Kim OY Kwon P Arvan (1996) ArticleTitleAn endoplasmic reticulum storage disease causing congenital goiter with hypothyroidism J Cell Biol 133 517–527 Occurrence Handle8636228 Occurrence Handle1:CAS:528:DyaK28XivVKnt7o%3D Occurrence Handle10.1083/jcb.133.3.517

    Article  PubMed  CAS  Google Scholar 

  3. SK DebBurman GJ Raymond B Caughey S Lindquist (1997) ArticleTitleChaperone-supervised conversion of prion protein to its protease-resistant form Proc Natl Acad Sci USA 94 13938–13943 Occurrence Handle9391131 Occurrence Handle1:CAS:528:DyaK2sXotVeht7Y%3D Occurrence Handle10.1073/pnas.94.25.13938

    Article  PubMed  CAS  Google Scholar 

  4. JD Harper PT Lansbury SuffixJr (1997) ArticleTitleModels of amyloid seeding in Alzheimer’s disease and scrapie: mechanistic truths and physiological consequences of the time-dependent solubility of amyloid proteins Annu Rev Biochem 66 385–407 Occurrence Handle9242912 Occurrence Handle1:CAS:528:DyaK2sXktFOhsrc%3D Occurrence Handle10.1146/annurev.biochem.66.1.385

    Article  PubMed  CAS  Google Scholar 

  5. CM Cabral Y Liu RN Sifers (2001) ArticleTitleDissecting glycoprotein quality control in the secretory pathway Trends Biochem Sci 26 619–624 Occurrence Handle11590015 Occurrence Handle1:CAS:528:DC%2BD3MXnt1Wjtrw%3D Occurrence Handle10.1016/S0968-0004(01)01942-9

    Article  PubMed  CAS  Google Scholar 

  6. T Kimura Y Hosoda Y Kitamura H Nakamura T Horibe M Kikuchi (2004) ArticleTitleFunctional differences between human and yeast protein disulfide isomerase family proteins Biochem Biophys Res Commun 320 359–365 Occurrence Handle15219835 Occurrence Handle1:CAS:528:DC%2BD2cXlt1egsbk%3D Occurrence Handle10.1016/j.bbrc.2004.05.178

    Article  PubMed  CAS  Google Scholar 

  7. L Ellgaard LW Ruddock (2005) ArticleTitleThe human protein disulphide isomerase family: substrate interactions and functional properties EMBO Rep 6 28–32 Occurrence Handle15643448 Occurrence Handle1:CAS:528:DC%2BD2MXitVersQ%3D%3D Occurrence Handle10.1038/sj.embor.7400311

    Article  PubMed  CAS  Google Scholar 

  8. R Günther C Brauer B Janetzky HH Forster IM Ehbrecht L Lehle H Küntzel (1991) ArticleTitleThe Saccharomyces cerevisiae TRG1 gene is essential for growth and encodes a lumenal endoplasmic reticulum glycoprotein involved in the maturation of vesicular carboxypeptidase J Biol Chem 266 24557–24563 Occurrence Handle1761554

    PubMed  Google Scholar 

  9. C Tachibana TH Stevens (1992) ArticleTitleThe yeast EUG1 gene encodes an endoplasmic reticulum protein that is functionally related to protein disulfide isomerase Mol Cell Biol 12 4601–4611 Occurrence Handle1406650 Occurrence Handle1:CAS:528:DyaK3sXks1Knsr0%3D

    PubMed  CAS  Google Scholar 

  10. H Tachikawa Y Takeuchi W Funahashi T Miura XD Gao D Fujimoto T Mizunaga K Onodera (1995) ArticleTitleIsolation and characterization of a yeast gene, MPD1, the overexpression of which suppresses inviability caused by protein disulfide isomerase depletion FEBS Lett 369 212–216 Occurrence Handle7649260 Occurrence Handle1:CAS:528:DyaK2MXnslyhsrY%3D Occurrence Handle10.1016/0014-5793(95)00750-4

    Article  PubMed  CAS  Google Scholar 

  11. H Tachikawa W Funahashi Y Takeuchi H Nakanishi R Nishihara S Katoh XD Gao T Mizunaga D Fujimoto (1997) ArticleTitleOverproduction of Mpd2p suppresses the lethality of protein disulfide isomerase depletion in a CXXC sequence dependent manner Biochem Biophys Res Commun 239 710–714 Occurrence Handle9367834 Occurrence Handle1:CAS:528:DyaK2sXnsVKksLY%3D Occurrence Handle10.1006/bbrc.1997.7426

    Article  PubMed  CAS  Google Scholar 

  12. R Xiao B Wilkinson A Solovyov JR Winther A Holmgren J Lundström-Ljung HF Gilbert (2004) ArticleTitleThe contribution of protein disulphide isomerase and its homologues to oxidative protein folding in the yeast endoplasmic reticulum J Biol Chem 279 49780–49786 Occurrence Handle15377672 Occurrence Handle1:CAS:528:DC%2BD2cXhtVWjs7%2FL Occurrence Handle10.1074/jbc.M409210200

    Article  PubMed  CAS  Google Scholar 

  13. A Solovyov R Xiao HF Gilbert (2004) ArticleTitleSulfhydryl oxidation, not disulfide isomerization, is the principal function of protein disulfide isomerase in yeast Saccharomyces cerevisiae J Biol. Chem 279 34095–34100 Occurrence Handle15175335 Occurrence Handle1:CAS:528:DC%2BD2cXmvFentr4%3D Occurrence Handle10.1074/jbc.M405640200

    Article  PubMed  CAS  Google Scholar 

  14. S Tanaka T Uehara Y Nomura (2000) ArticleTitleUp-regulation of protein-disulfide isomerase in response to hypoxia/brain ischemia and its protective effect against apoptotic cell death J Biol Chem 275 10388–20393 Occurrence Handle10744727 Occurrence Handle1:CAS:528:DC%2BD3cXisFWjt78%3D Occurrence Handle10.1074/jbc.275.14.10388

    Article  PubMed  CAS  Google Scholar 

  15. M Saloheimo M Lund ME Penttila (1999) ArticleTitleThe protein disulphide isomerase gene of the fungus Trichoderma reesei is induced by endoplasmic reticulum stress and regulated by the carbon source Mol Gen Genet 262 354–5

    Google Scholar 

  16. DJ Jeenes R Pfaller DB Archer (1997) ArticleTitleIsolation and characterization of a novel stress-inducible PDI-family gene from Aspergillus niger Gene 193 151–156 Occurrence Handle9256071 Occurrence Handle1:CAS:528:DyaK2sXks12gtrw%3D Occurrence Handle10.1016/S0378-1119(97)00098-X

    Article  PubMed  CAS  Google Scholar 

  17. M Goecke C Gallant P Suntharalingam NL Martin (2002) ArticleTitleSalmonella typhimurium DsbA is growth-phase regulated FEMS Microbiol Lett 206 229–234 Occurrence Handle11814668 Occurrence Handle1:CAS:528:DC%2BD38Xos1OksQ%3D%3D Occurrence Handle10.1111/j.1574-6968.2002.tb11014.x

    Article  PubMed  CAS  Google Scholar 

  18. AM Myers A Tzagoloff DM Kinney CJ Lusty (1986) ArticleTitleYeast shuttle and integrative vectors with multiple cloning sites suitable for construction of lacZ fusions Gene 45 299–310 Occurrence Handle3026915 Occurrence Handle1:CAS:528:DyaL2sXksV2kug%3D%3D Occurrence Handle10.1016/0378-1119(86)90028-4

    Article  PubMed  CAS  Google Scholar 

  19. NA Morjana HF Gilbert (1991) ArticleTitleEffect of protein and peptide inhibitors on the activity of protein disulfide isomerase Biochemistry 30 4985–4990 Occurrence Handle2036365 Occurrence Handle1:CAS:528:DyaK3MXit1Cgsbw%3D Occurrence Handle10.1021/bi00234a021

    Article  PubMed  CAS  Google Scholar 

  20. L Guarente (1983) ArticleTitleYeast promoters and lacZ fusions designed to study expression of cloned genes in yeast Methods Enzymol 101 181–191 Occurrence Handle6310321 Occurrence Handle1:CAS:528:DyaL2cXhvVOhsA%3D%3D Occurrence Handle10.1016/0076-6879(83)01013-7

    Article  PubMed  CAS  Google Scholar 

  21. MM Bradford (1976) ArticleTitleA rapid and sensitive method for the quantitation of microgram quantities of protein utilizing the principle of protein-dye binding Biochem 72 248–254 Occurrence Handle1:CAS:528:DyaE28XksVehtrY%3D

    CAS  Google Scholar 

  22. DP Dixon M Van Lith R Edwards A Benham (2003) ArticleTitleCloning and initial characterization of the Arabidopsis thaliana endoplasmic reticulum oxidoreductins Antioxid Redox Signal 5 389–396 Occurrence Handle13678526 Occurrence Handle1:CAS:528:DC%2BD3sXmvFCmtbo%3D Occurrence Handle10.1089/152308603768295122

    Article  PubMed  CAS  Google Scholar 

  23. XD Liu DJ Thiele (1997) ArticleTitleYeast metallothionein gene expression in response to metals and oxidative stress Methods 11 289–299 Occurrence Handle9073572 Occurrence Handle1:CAS:528:DyaK2sXhvFeku7Y%3D Occurrence Handle10.1006/meth.1996.0423

    Article  PubMed  CAS  Google Scholar 

  24. HF Marshall K Merchant JS Stamler (2000) ArticleTitleNitrosation and oxidation in the regulation of gene expression FASEB J 14 1889–1900 Occurrence Handle11023973 Occurrence Handle1:CAS:528:DC%2BD3MXhtF2rs7s%3D Occurrence Handle10.1096/fj.00.011rev

    Article  PubMed  CAS  Google Scholar 

  25. C Nathan MU Shiloh (2000) ArticleTitleReactive oxygen and nitrogen intermediates in the relationship between mammalian hosts and microbial pathogens Proc Natl Acad Sci USA 97 8841–8848 Occurrence Handle10922044 Occurrence Handle1:CAS:528:DC%2BD3cXls12ltLo%3D Occurrence Handle10.1073/pnas.97.16.8841

    Article  PubMed  CAS  Google Scholar 

  26. F Rolland J Winderickx JM Thevelein (2001) ArticleTitleGlucose-sensing mechanisms in eukaryotic cells Trends Biochem Sci 26 310–317 Occurrence Handle11343924 Occurrence Handle1:CAS:528:DC%2BD3MXjtlCjt7g%3D Occurrence Handle10.1016/S0968-0004(01)01805-9

    Article  PubMed  CAS  Google Scholar 

  27. DR Spitz JE Sim LA Ridnour SS Galoforo YJ Lee (2000) ArticleTitleGlucose deprivation-induced oxidative stress in human tumor cells: A fundamental defect in metabolism Ann NY Acad Sci 899 349–362 Occurrence Handle10863552 Occurrence Handle1:CAS:528:DC%2BD3cXks1Shsbw%3D Occurrence Handle10.1111/j.1749-6632.2000.tb06199.x

    Article  PubMed  CAS  Google Scholar 

  28. YJ Lee SS Galoforo CM Berns JC Chen BH Davis JE Sim PM Corry DR Spitz (1998) ArticleTitleGlucose deprivation-induced cytotoxicity and alterations in mitogen-activated protein kinase activation are mediated by oxidative stress in multidrug-resistant human breast carcinoma Cells J Biol Chem 273 5294–5299 Occurrence Handle9478987 Occurrence Handle1:CAS:528:DyaK1cXhs1CnsLY%3D Occurrence Handle10.1074/jbc.273.9.5294

    Article  PubMed  CAS  Google Scholar 

  29. JJ Song JG Rhee M Suntharalingam SA Walsh DR Spitz YJ Lee (2002) ArticleTitleRole of glutaredoxin in metabolic oxidative stress Glutaredoxin as a sensor of oxidative stress mediated by H2O2 J Biol Chem 277 46566–46575 Occurrence Handle12244106 Occurrence Handle1:CAS:528:DC%2BD38XovFKgsrw%3D Occurrence Handle10.1074/jbc.M206826200

    Article  PubMed  CAS  Google Scholar 

  30. S Stettler E Warbrick S Prochnik S Mackie P Fantes (1996) ArticleTitleThe wis1 signal transduction pathway is required for expression of cAMP-repressed genes in fission yeast J Cell Sci 109 1927–1935 Occurrence Handle8832415 Occurrence Handle1:CAS:528:DyaK28XksFCktrs%3D

    PubMed  CAS  Google Scholar 

  31. K Shiozaki M Shiozaki P Russell (1997) ArticleTitleMcs4 mitotic catastrophe suppressor regulates the fission yeast cell cycle through the Wik1-Wis1-Spc1 kinase cascade Mol Biol Cell 8 409–419 Occurrence Handle9188094 Occurrence Handle1:CAS:528:DyaK2sXit1yrs7s%3D

    PubMed  CAS  Google Scholar 

  32. M Madrid T Soto A Franco V Paredes E Hidalgo M Gracto J Cansado (2004) ArticleTitleA cooperative role for Atf1 and Pap1 in the detoxification of the oxidative stress induced by glucose deprivation in Schizosaccharomyces pombe J Biol Chem 279 41594–41602 Occurrence Handle15247218 Occurrence Handle1:CAS:528:DC%2BD2cXnvFWgs7k%3D Occurrence Handle10.1074/jbc.M405509200

    Article  PubMed  CAS  Google Scholar 

  33. M Hiesinger S Roth E Meissner HJ Schuller (2001) ArticleTitleContribution of Cat8 and Sip4 to the transcriptional activation of yeast gluconeogenic genes by carbon source-responsive elements Curr Genet 39 68–76 Occurrence Handle11405098 Occurrence Handle1:CAS:528:DC%2BD3MXivFaisrg%3D Occurrence Handle10.1007/s002940000182

    Article  PubMed  CAS  Google Scholar 

  34. K Mehdi J Thierie MJ Penninckx (2001) ArticleTitleγ-Glutamyl transpeptidase in the yeast Saccharomyces cerevisiae and its role in the vacuola transport and metabolism of glutathione Biochem J 359 631–637 Occurrence Handle11672438 Occurrence Handle1:CAS:528:DC%2BD3MXovVSisb8%3D Occurrence Handle10.1042/0264-6021:3590631

    Article  PubMed  CAS  Google Scholar 

  35. J-Y Springael MJ Penninckx (2003) ArticleTitleNitrogen-source regulation of yeast-glutamyl transpeptidase synthesis involves the regulatory network including the GATA zinc-finger factor Gln3, Nil1/Gat1 and Gzf3 Biochem J 371 589–595 Occurrence Handle12529169 Occurrence Handle1:CAS:528:DC%2BD3sXisleqsLw%3D Occurrence Handle10.1042/BJ20021893

    Article  PubMed  CAS  Google Scholar 

  36. Y-Y Lee S-J Kim E-H Park C-J Lim (2003) ArticleTitleGlutathione content and the activities of glutathione synthesizing enzymes in fission yeast are modulated by oxidative stress J Microbiol 41 248–251 Occurrence Handle1:CAS:528:DC%2BD3sXosFClt74%3D

    CAS  Google Scholar 

  37. AN Nguyen A Lee W Place K Shiozaki (2000) ArticleTitleMultistep phosphorelay proteins transmit oxidative stress signals to the fission yeast stress-activated protein kinase Mol Biol Cell 11 1169–1181 Occurrence Handle10749922 Occurrence Handle1:CAS:528:DC%2BD3cXlt1Wlu7o%3D

    PubMed  CAS  Google Scholar 

  38. WM Toone S Kuge M Samuels BA Morgan T Toda N Jones (1998) ArticleTitleRegulation of the fission yeast transcription factor Pap1 by oxidative stress: requirement for the nuclear export factor Crm1 (exportin) and the stress-activated MAP kinase Sty1/Spc1 Genes Dev 12 1453–1463 Occurrence Handle9585505 Occurrence Handle1:CAS:528:DyaK1cXjtlKrsbw%3D

    PubMed  CAS  Google Scholar 

  39. Y Fujii T Shimizu T Toba M Yanagida T Hakoshima (2000) ArticleTitleStructural basis for the diversity of DNA recognition by bZIP transcription factors Nat Struct Biol 7 889–893 Occurrence Handle11017199 Occurrence Handle1:CAS:528:DC%2BD3cXns1eiu7c%3D Occurrence Handle10.1038/82822

    Article  PubMed  CAS  Google Scholar 

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Authors and Affiliations

  1. Division of Life Sciences, College of Natural Sciences, Kangwon National University, 192-1 Hyoja-2-dong, Chuncheon, 200-701, Korea

    Su-Jung Kim, Yeon-Sook Choi, Hong-Gyum Kim & Chang-Jin Lim

  2. College of Pharmacy, Sookmyung Women’s University, Seoul, 140-742, Korea

    Eun-Hee Park

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  1. Su-Jung Kim
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  2. Yeon-Sook Choi
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  3. Hong-Gyum Kim
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  4. Eun-Hee Park
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Correspondence to Chang-Jin Lim.

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Kim, SJ., Choi, YS., Kim, HG. et al. Cloning, characterization and regulation of a protein disulfide isomerase from the fission yeast Schizosaccharomyces pombe . Mol Biol Rep 33, 187–196 (2006). https://doi.org/10.1007/s11033-006-0012-9

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  • DOI: https://doi.org/10.1007/s11033-006-0012-9

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