Abstract
Protein disulfide isomerase (PDI) is a thiodisulfide oxidoreductase that catalyzes the formation, reduction and rearrangement of disulfide bonds in proteins of eukaryotes. The classical PDI has a signal peptide, two CXXCcontaining thioredoxin catalytic sites (a,a′), two noncatalytic thioredoxin fold domains (b,b′), an acidic domain (c) and a C-terminal endoplasmic reticulum (ER) retention signal. Although PDI resides in the ER where it mediates the folding of nascent polypeptides of the secretory pathway, we recently showed that PDI5 of Arabidopsis thaliana chaperones and inhibits cysteine proteases during trafficking to vacuoles prior to programmed cell death of the endothelium in developing seeds. Here we describe Arabidopsis PDI2, which shares a primary structure similar to that of classical PDI. Recombinant PDI2 is imported into ER-derived microsomes and complements the E. coli protein-folding mutant, dsbA. PDI2 interacted with proteins in both the ER and nucleus, including ER-resident protein folding chaperone, BiP1, and nuclear embryo transcription factor, MEE8. The PDI2-MEE8 interaction was confirmed to occur in vitro and in vivo. Transient expression of PDI2-GFP fusions in mesophyll protoplasts resulted in labeling of the ER, nucleus and vacuole. PDI2 is expressed in multiple tissues, with relatively high expression in seeds and root tips. Immunoelectron microscopy with GFP- and PDI2-specific antisera on transgenic seeds (PDI2-GFP) and wild type roots demonstrated that PDI2 was found in the secretory pathway (ER, Golgi, vacuole, cell wall) and the nuclei. Our results indicate that PDI2 mediates protein folding in the ER and has new functional roles in the nucleus.
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Adikesavan, A., Karani, U., Emmanual, J., and Anil, K. (2008). Overlapping signal sequences control nuclear localization and endoplasmic reticulum retention of GRP58. Biochem. Biophys. Res. Commun. 377, 407–412.
Amaya, Y., Nakai, T., Komaru, K., Tsuneki, M., and Miura, S. (2008). Cleavage of the ER-targeting signal sequence of parathyroid hormone-related protein is cell-type-specific and regulated in cis by its nuclear localization signal. J. Biochem. 143, 569–579.
Armstrong, D.J., and Roman, A. (1993). The anomalous electrophoretic behavior of the human papillomavirus type 16 E7 protein is due to the high content of acidic amino acid residues. Biochem. Biophys. Res. Commun. 192, 1380–1387.
Asally, M., and Yoneda, Y. (2005). Beta-catenin can act as a nuclear import receptor for its partner transcription factor, lymphocyte enhancer factor-1 (lef-1). Exp. Cell. Res. 308, 357–363.
Aslund, F., and Beckwith, J. (1999). Bridge over troubled waters: sensing stress by disulfide bond formation. Cell 96, 751–753.
Battey, N.H., James, N.C., Greenland, A.J., and Brownlee, C. (1999). Exocytosis and Endocytosis. Plant Cell 11, 643–659.
Bendtsen, J.D., Nielsen, H., von Heijne, G., and Brunak, S. (2004). Improved prediction of signal peptides: SignalP 30. J. Mol. Biol. 340, 783–795.
Bertolotti, A., Zhang, Y., Hendershot, L.M., Harding, H.P., and Ron, D. (2000). Dynamic interaction of BiP and ER stress transducers in the unfolded-protein response. Nat. Cell. Biol. 2, 326–332.
Bleckmann, A., Weidtkamp-Peters, S., Seidel, C.A., and Simon, R. (2010). Stem cell signaling in Arabidopsis requires CRN to localize CLV2 to the plasma membrane. Plant Physiol. 152, 166–176.
Cheug, P.Y., and Churchich, J.E. (1999). Recognition of protein substrates by protein-disulfide isomerase A sequence of the b′ domain responds to substrate binding. J. Biol. Chem. 274, 32757–32761.
Christopher, D.A., Borsics, T., Yuen, C.Y., Ullmer, W., Andème-Ondzighi, C., Andres, M.A., Kang, B.H., and Staehelin, L.A. (2007). The cyclic nucleotide gated cation channel AtCNGC10 traffics from the ER via Golgi vesicles to the plasma membrane of Arabidopsis root and leaf cells. BMC Plant Biol. 7, 1471–2229.
Chun, L., Kawakami, A., and Christopher, D.A. (2001). Phytochrome A mediates blue light and UV-A dependent chloroplast gene transcription in green leaves. Plant Physiol. 125, 1957–1966.
Clive, D.R., and Greene, J.J. (1996). Cooperation of protein disulfide isomerase and redox environment in the regulation of NF-κB and AP1 binding to DNA. Cell Biochem. Funct. 14, 49–55.
Clough, S., and Bent, A. (1998). Floral dip: a simplified method for Agrobacterium-mediated transformation of Arabidopsis thaliana. Plant J. 16, 735–743.
Cokol, M., Nair, R., and Rost, B. (2000). Finding nuclear localisation signals. EMBO Rep. 1, 411–415.
Couet, J., De Bernard, S., Loosfelt, H., Saunier, B., Milgrom, E., and Misrahi, M. (1996). Cell surface protein disulfide-isomerase is involved in the shedding of human thyrotropin receptor ectodomain. Biochemistry 35, 14800–14805.
Cramer, J.H., Lea, K., Schaber, M.D., and Kramer, R.A. (1987). Signal peptide specificity in post-translational processing of the plant protein phaseolin in Saccharomyces cerevisiae. Mol. Cell. Biol. 7, 121–128.
Durfee, T., Becherer, K., Chen, P.L., Yeh, S.H., Yang, Y., Kilburn, A.E., Lee, W.H., and Elledge, S.J. (1993). The retinoblastoma protein associates with the protein phosphatase type 1 catalytic subunit. Genes Dev. 7, 555–569.
Favaloro, V., Spasic, M., Schwappach, B., and Dobberstein, B. (2008). Distinct targeting pathways for the membrane insertion of tail-anchored (TA). proteins. J. Cell Sci. 121, 1832–1840.
Ferrari, D.M., and Soling, H.D. (1999). The protein disulphide-isomerase family: unraveling a string of folds. Biochem. J. 339, 1–10.
Fields, S., and Song, O. (1989). A novel genetic system to detect protein-protein interactions. Nature 20, 245–246.
Finn, R.D., Mistry, J., Schuster-Böckler, B., Griffiths-Jones, S., Hollich, V., Lassmann, T., Moxou, S., Marshall, M., Khanna, A., Durbin, R., et al. (2006). Pfam: clans, web tools and services. Nucleic Acids Res. 34, D247–D251.
Frand, A.R., and Kaiser, C.A. (1998). The ERO1 gene of yeast is required for oxidation of protein dithiols in the endoplasmic reticulum. Mol. Cell 1, 161–170.
Furumizu, C., Tsukaya, H., and Komeda, Y. (2010). Characterization of EMU, the Arabidopsis homolog of the yeast THO complex member HPR1. RNA 16, 1809–1817.
Gruber, C.W., Cemazar, M., Heras, B., Martin, J.L., and Craik, D.J. (2006). Protein disulfide isomerase: the structure of oxidative folding. Trends Biochem. Sci. 31, 455–464.
Gruber, C.W., Cemazar, M., Clark, R.J.T., Renda, R.F., Anderson, M.A., and Craik, D.J. (2007). A novel plant protein-disulfide isomerase involved in the oxidative folding of cystine knot defense proteins. J. Biol. Chem. 282, 20435–20446.
Haseloff, J., Siemering, K.R., Prasher, D.C., and Hodge, S. (1997). Removal of a cryptic intron and subcellular localization of green fluorescent protein are required to mark transgenic Arabidopsis plants brightly. Proc. Natl. Acad. Sci. USA 94, 2122–2127.
Helm, M., Schmid, M., Hierl, G., Terneus, K., Tan, L., Lottspeich, F., Kieliszewski, M.J., and Gietl, C. (2008). KDEL-tailed cysteine endopeptidases involved in programmed cell death, intercalation of new cells, and dismantling of extensin scaffolds. Am. J. Bot. 95, 1049–1062.
Higuchi, T., Watanabe, Y., and Waga, I. (2004). Protein disulfide isomerase suppresses the transcriptional activity of NF-κB. Bio chem. Biophys. Res. Commun. 318, 46–52.
Holst, B., Tachibana, C., and Winther, J.R. (1997). Active site mutations in yeast protein disulfide isomerase cause dithiothreitol densitivity and a reduced rate of protein folding in the endoplasmic reticulum. J. Cell. Biol. 138, 1229–1238.
Honscha, W., Ottallah, M., Kistner, A., Platte, H., and Petzinger, E. (1993). A membrane-bound form of protein disulfide isomerase (PDI). and the hepatic uptake of organic anions. Biochim. Biophys. Acta 1153, 175–183.
Houston, N.L., Fan, C., Xiang, J.Q., Schulze, J.M., Jung, R., and Boston, R.S. (2005). Phylogenetic analyses identify 10 classes of the protein disulfide isomerase family in plants, including single-domain protein disulfide isomerase-related proteins. Plant Physiol. 137, 762–778.
Jackson, R.C., and Blobel, G. (1977). Post-translational cleavage of presecretory proteins with an extract of rough microsomes from dog pancreas containing signal peptidase activity. Proc. Natl. Acad. Sci. USA 74, 5598–5602.
James, P., Halladay, J., and Craig, E.A. (1996). Genomic libraries and a host strain designed for highly efficient two-hybrid selection in yeast. Genetics 144, 1425–1436.
Jauvion, V., Elmayan, T., and Vaucheret, H. (2010). The conserved RNA trafficking proteins HPR1 and TEX1 are involved in the production of endogenous and exogenous small interfering RNA in Arabidopsis. Plant Cell 22, 2697–2709.
John, D.C., and Bulleid, N.J. (1994). Prolyl 4-hydroxylase: defective assembly of alpha-subunit mutants indicates that assembled alpha-subunits are intramolecularly disulfide bonded. Biochemistry 33, 14018–14025.
Jones, A.M., and Herman, E.M. (1993). A K-D-E-L-containing auxin-binding protein is located at the plasma membrane and within the cell wall. Plant Physiol. 101, 595–606.
Kanai, S., Toh, H., Hayano, T., and Kikuchi, M. (1998). Molecular evolution of the domain structures of protein disulfide isomerases. J. Mol. Evol. 47, 200–210.
Karniely, S., and Pines, O. (2005). Single translation-dual destination: mechanisms of dual protein targeting in eukaryotes. EMBO Rep. 6, 420–425.
Kerk, D., Bulgrien, J., Smith, D.W., and Gribskov, M. (2003). Arabidopsis proteins containing similarity to the universal stress protein domain of bacteria. Plant Physiol. 131, 1209–1219.
Kim, J.M., and Mayfield, S.P. (1997). Protein disulfide isomerase as a regulator of chloroplast translational activation. Science 278, 1954–1957.
Kong, Z., Li, M., Yang, W., Xu, W., and Xue, Y. (2006). A novel nuclear-localized CCCH-type zinc finger protein, OsDOS, is involved in delaying leaf senescence in rice. Plant Physiol. 141, 1376–1388.
Lahav, J., Gofer-Dadosh, N., Luboshitz, J., Hess, O., and Shaklai, M. (2000). Protein disulfide isomerase mediates integrindependent adhesion. FEBS Lett. 475, 89–92.
Lamberg, A., Jauhiainen, M., Metso, J., Ehnholm, C., Shoulders, C., Scott, J., Pihlajaniemi, T., and Kivirikko, K.I. (1996). The role of protein disulphide isomerase in the microsomal triacylglycerol transfer protein does not reside in its isomerase activity. Biochem. J. 315, 533–536.
Lange, A., Mills, R.E., Lange, C.J., Stewart, M., Devine, S.E., and Corbett, A.H. (2007). Classical nuclear localization signals: definition, function, and interaction with importin alpha. J. Biol. Chem. 282, 5101–5105.
Lappi, A.K., Lensink, M.F., Alanen, H.I., Salo, K.E., Lobell, M., Juffer, A.H., and Ruddock, L.W. (2004). A conserved arginine plays a role in the catalytic cycle of the protein disulphide isomerases. J. Mol. Biol. 335, 283–295.
Lee, S.-O., Cho, K., Cho, S., Kim, I., Oh, C., and Ahn, K. (2010). Protein disulphide isomerase is required for signal peptide peptidase-mediated protein degradation. EMBO J. 29, 363–375.
Levitan, A., Trebitsh, T., Kiss, V., Pereg, Y., Dangoor, I., and Danon, A. (2005). Dual targeting of the protein disulfide isomerase RB60 to the chloroplast and the endoplasmic reticulum. Proc. Natl. Acad. Sci. USA 102, 6225–6230.
Li, C.P., and Larkins, B.A. (1996). Expression of protein disulfide isomerase is elevated in the endosperm of the maize floury 2 mutant. Plant Mol. Biol. 30, 873–882.
Lu, D.-P., and Christopher, D.A. (2006). Immunolocalization of a protein disulfide isomerase to Arabidopsis thaliana chloroplasts and its association with starch biogenesis. Int. J. Plant Sci. 167, 1–9.
Lu, D.-P., and Christopher, D.A. (2008). Endoplasmic reticulum stress activates the expression of a sub-group of protein disulfide isomerase genes and AtbZIP60 modulates the response in Arabidopsis thaliana. Mol. Genet. Genomics 280, 199–210.
Lucero, H.A., and Kaminer, B. (1999). The role of calcium on the activity of ER calcistorin/Protein-disulfide Isomerase and the significance of the C-terminal and itscalcium binding A comparison with mammalian protein-disulfide isomerase. J. Biol. Chem. 274, 3243–3251.
Lumb, R.A., and Bulleid, N.J. (2002). Is protein disulfide isomerase a redox-dependent molecular chaperone? EMBO J. 21, 6763–6770.
Markus, M., and Benezra, R. (1999). Two isoforms of protein disulfide isomerase alter the dimerization status of E2A proteins by a redox mechanism. J. Biol. Chem. 274, 1040–1049.
Maruyama, D., Endo, T., and Nishikawa, S. (2010). BiP-mediated polar nuclei fusion is essential for the regulation of endosperm nuclei proliferation in Arabidopsis thaliana. Proc. Natl. Acad. Sci. USA 107, 1684–1689.
Mayer, M., Kies, U., Kammermeier, R., and Buchner, J. (2000). BiP and PDI cooperate in the oxidative folding of antibodies in vitro. J. Biol. Chem. 275, 29421–29425.
Motohashi, K., Kondoh, A., Stumpp, M.T., and Hisabori, T. (2001). Comprehensive survey of proteins targeted by chloroplast thioredoxin. Proc. Natl. Acad. Sci. USA 98, 11224–11229.
Nelson, B.K., Cai, X., and Nebenführ, A. (2007). A multicolored set of in vivo organelle markers for co-localization studies in Arabidopsis and other plants. Plant J. 51, 1126–1136.
Neuteboom, L.W., Matsumoto, K.O., and Christopher, D.A. (2009). An extended AE-rich N-terminal trunk in secreted pineapple cystatin enhances inhibition of fruit bromelain and is post-translationally removed during ripening. Plant Physiol. 151, 515–527.
Ohtani, H., Wakui, H., Ishino, T., Komatsuda, A., and Miura, A.B. (1993). An isoform of protein disulfide isomerase is expressed in the developing acrosome of spermatids during rat spermiogenesis and is transported into the nucleus of mature spermatids and epididymal spermatozoa. Histochemistry 100, 423–429.
Okita, T.W., and Rogers, J.C. (1996). Compartmentation of proteins in the endomembrane system of plant cells. Annu. Rev. Plant. Physiol. Plant Mol. Biol. 47, 327–350.
Onda, Y., Nagamine, A., Sakurai, M., Kumamaru, T., Ogawa, M., and Kawagoe, Y. (2011). Distinct roles of protein disulfide isomerase and p5 sulfhydryl oxidoreductases in multiple pathways for oxidation of structurally diverse storage proteins in rice. Plant Cell 23, 210–223.
Ondzighi, C.A., Christopher, D.A., Cho, E.J., Chang, S.C., and Staehelin, L.A. (2008). Arabidopsis protein disulfide isomerase-5 inhibits cysteine proteases during trafficking to vacuoles before programmed cell death of the endothelium in developing seeds. Plant Cell 20, 2205–2220.
Ostermeier, M., De Sutter, K., and Georgiou, G. (1996). Eukaryotic protein disulfide isomerase complements E. coli dsbA mutants and increases the yield of heterologous secreted protein with disulfide bonds. J. Biol. Chem. 271, 10616–10622.
Pagnussat, G.C., Yu, H.J., Ngo, Q.A., Rajani, S., Mayalagu, S., Johnson, C.S., Capron, A., Xie, L.F., Ye, D., and Sundaresan, V. (2005). Genetic and molecular identification of genes required for female gametophyte development and function in Arabidopsis. Development 132, 603–614.
Plath, K., Mothes, W., Wilkinson, B.M., Stirling, C.J., and Rapoport, T.A. (1998). Signal sequence recognition in posttranslational protein transport across the yeast ER membrane. Cell 94, 795–807.
Pomeranz, M.C., Hah, C., Lin, P.-C., Kang, S.G., Finer, J.J., Blackshear, P.J., and Jang, J.C. (2010). The Arabidopsis tandem zinc finger protein AtTZF1 traffics between the nucleus and cytoplasmic foci and binds both DNA and RNA. Plant Physiol. 152, 151–165.
Powers, T., and Walter, P. (1996). The nascent polypeptide-associated complex modulates interactions between the signal recognition particle and the ribosome. Curr. Biol. 6, 331–338.
Rapoport, T.A., Matlack, K.E., Plath, K., Misselwitz, B., and Staeck, O. (1999). Posttranslational protein translocation across the membrane of the endoplasmic reticulum. Biol. Chem. 380, 1143–1150.
Rigobello, M.P., Donella-Deana, A., Cesaro, L., and Bindoli, A. (2001). Distribution of protein disulphide isomerase in rat liver mitochondria. Biochem. J. 356, 567–570.
Rosenthal, J.A., Chen, H., Slepnev, V.I., Pellegrini, L., Salcini, A.E., Di Fiore, P.P., and De Camilli, P. (1999). The epsins define a family of proteins that interact with components of the clathrin coat and contain a new protein module. J. Biol. Chem. 274, 33959–33965.
Rout, M.P., Aitchison, J.D., Magnasco, M.O., and Chait, B.T. (2003). Virtual gating and nuclear transport: the hole picture. Trends Cell Biol. 13, 622–628.
Shimada, H., Mochizuki, M., Ogura, K., Froehlich, J.E., Osteryoung, K.W., Shirano, Y., Shibata, D., Masuda, S., Mori, K., and Takamiya, K. (2007). Arabidopsis cotyledon-specific chloroplast biogenesis factor CYO1 is a protein disulfide isomerase. Plant Cell 19, 3157–3169.
Shimoni, Y., Zhu, X.Z., Levanony, H., Segal, G., and Galili, G. (1995). Purification characterization and intracellular localization of glycosylated protein disulfide isomerase from wheat grains. Plant Physiol. 108, 327–335.
Sone, M., Kishigami, S., Yoshihisa, T., and Ito, K. (1997). Roles of disulfide bonds in bacterial alkaline phosphatase. J. Biol. Chem. 272, 6174–6178.
Stockton, J.D., Merkert, M.C., and Kellaris, K.V. (2003). A complex of chaperones and disulfide isomerases occludes the cytosolic face of the translocation protein Sec61p and affects translocation of the prion protein. Biochemistry 42, 12821–12834.
Swanson, R., Locher, M., and Hochstrasser, M. (2001). A conserved ubiquitin ligase of the nuclear envelope/endoplasmic reticulum that functions in both ER-associated and matalpha2 repressor degradation. Genes Dev. 15, 2660–2674.
Takemoto, Y., Coughlan, S.J., Okita, T.W., Satoh, H., Ogawa, M., and Kumamaru, T. (2002). The rice mutant esp2 greatly accumulates the glutelin precursor and deletes the protein disulfide isomerase. Plant Physiol. 128, 1212–1222.
Tamura, K., Yamada, K., Shimada, T., and Hara-Nishimura, I. (2004). Endoplasmic reticulum-resident proteins are constitutively transported to vacuoles for degradation. Plant J. 39, 393–402.
Tu, B., Ho-Schleyer, S.C., Travers, K.J., and Weissman, K.J. (2000). Biochemical basis of oxidative protein folding in the endoplasmic reticulum. Science 290, 1571–1574.
Turano, C., Coppari, S., Altieri, F., and Ferraro, A. (2002). Proteins of the PDI family: unpredicted non-ER locations and functions. J. Cell. Physiol. 193, 154–163.
Wang, H., Boavida, L.C., Ron, M., and McCormick, S. (2008). Truncation of a protein disulfide isomerase, PDIL2-1, delays embryo sac maturation and disrupts pollen tube guidance in Arabidopsis thaliana. Plant Cell 20, 3300–3311.
Wilson, R., Lees, J.F., and Bulleid, N.J. (1998). Protein disulfide isomerase acts as a molecular chaperone during the assembly of procollagen. J. Biol. Chem. 273, 9637–9643.
Wu, F.H., Shen, S.C., Lee, L.Y., Lee, S.H., Chan, M.T., and Lin, C. -S. (2009). Tape-Arabidopsis sandwich — a simpler Arabidopsis protoplast isolation method. Plant Methods 5, 1–10.
Xu, P., Radena, D., Doyle, III F.J., and Robinson, A.S. (2005). Analysis of unfolded protein response during single-chain antibody expression in Saccaromyces cerevisiae reveals different roles for BiP and PDI in folding. Metab. Eng. 7, 269–279.
Yoo, S.D., Cho, Y.H., and Sheen, J. (2007). Arabidopsis mesophyll protoplasts: a versatile cell system for transient gene expression analysis. Nat. Protoc. 2, 1565–1572.
Yoshimori, T., Semba, T., Takemoto, H., Akagi, S., Yamamoto, A., and Tashiro, Y. (1990). Protein disulfide-isomerase in rat exocrine pancreatic cells is exported from the endoplasmic reticulum despite possessing the retention signal. J. Biol. Chem. 265, 15984–15990.
Zaltsman, A., Yi, B.Y., Gafni, Y., and Citovsky, V. (2007). Yeastplant coupled vector system for identification of nuclear proteins. Plant Physiol. 145, 1264–1271.
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Cho, E.J., Yuen, C.Y., Kang, BH. et al. Protein disulfide isomerase-2 of Arabidopsis mediates protein folding and localizes to both the secretory pathway and nucleus, where it interacts with maternal effect embryo arrest factor. Mol Cells 32, 459–475 (2011). https://doi.org/10.1007/s10059-011-0150-3
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DOI: https://doi.org/10.1007/s10059-011-0150-3