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Is a ferroxidase involved in the high-affinity iron uptake in Chlamydomonas reinhardtii?

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Abstract

In contrast to the halotolerant algae Dunaliella salina, Fe assimilation in the unicellular green algae Chlamydomonas reinhardtii involves a reductive step. After fractionation of proteins from Fe-sufficient and Fe-deficient cells, a distinct increase of a 150 kDa protein was observed in the Fe-deficient plasma membrane. The induction of the Fe3+ reductase activity and the 150 kDa protein under Fe deficiency was completely blocked by cycloheximide, indicating transcriptional regulation. After tryptic digestion, internal peptide sequences were obtained using MALDI-TOF MS. Based on the homology of these sequences to human hephaestin, a multicopper oxidase, which is involved in Fe homeostasis, the 150 kDa protein was named: ferroxidase-like protein (FLP). An oxidase activity measured as p-phenylenediamine oxidation, was also increased under Fe-deficiency, and this oxidase was inhibited by the inhibitor for multicopper oxidases, tetrathiomolybdate. Based on the physiological experiments and the homology in the primary structure of the ferroxidase-like protein to multicopper oxidases, we suggest that as in yeast and humans an oxidase is involved in the Fe uptake in C. reinhardtii.

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References

  • Askwith C and Kaplan J 1998 Site-directed mutagenesis of the yeast multicopper oxidase Fet3p. J. Biol. Chem. 273, 22415-22419.

    Google Scholar 

  • Askwith C, Eide D, Van Ho A, Bernard P S, Li L, Davis-Kaplan S, Sipe D M and Kaplan J 1994 The FET3 gene of S. cerevisiae encodes a multicopper oxidase required for ferrous iron uptake. Cell 76, 403-410.

    Google Scholar 

  • Attieh Z K, Mukhopadhyay C K, Seshadri V, Tripoulas N A and Fox P L 1999 Ceruloplasmin ferroxidase activity stimulates cellular iron uptake by a trivalent cation-specific transport mechanism. J. Biol. Chem. 274, 1116-1123.

    Google Scholar 

  • Bonaccorsi di Patti M C, Felice M R, Camuti A P, Lania A and Musci G 2000 The essential role of Glu-185 and Tyr-354 residues in the ferroxidase activity of Saccharomyces cerevisiae Fet3. FEBS 472, 283-286.

    Google Scholar 

  • Buckhout T J, Luster D G and Chaney R L 1989 Iron-stress induced redox activity in tomato (Lycopersicon esculentum Mill.) is localized on the plasma membrane. Plant Physiol. 90, 151-156.

    Google Scholar 

  • Bradford M M 1976 A rapid and sensitive method for the quantitation of microgram quantities of protein utilizing the principle of protein-dye binding. Anal. Biochem. 72, 248-254.

    Google Scholar 

  • Chaney R L, Brown J C and Tiffin L O 1972 Obligatory reduction of ferric chelates in iron uptake by soybeans. Plant Physiol. 50, 208-213.

    Google Scholar 

  • Chidambaram M V, Barnes G and Frieden E 1984 Inhibition of ceruloplasmin and other copper oxidases by thiomolybdate. J. Inorg. Biochem. 22, 231-239.

    Google Scholar 

  • Curie C, Alonso J M, Le Jean M, Ecker J R and Briat J F 2000 Involvement of NRAMP 1 from Arabidopsis thaliana in iron transport. Biochem. J. 3, 749-755.

    Google Scholar 

  • Dancis A, Klausner R D, Hinnebusch A G and Barriocanal J G 1990 Genetic evidence that ferric reduction is required for iron uptake in Saccharomyces cerevisiae. Mol. Cell. Biol. 10, 2294-2301.

    Google Scholar 

  • Dancis A, Roman D G, Anderson G J and Klausner R D 1992 Ferric reductase of Saccharomyces cerevisiae: molecular characterization, role in iron uptake and transcriptional control by iron. Proc. Natl. Acad. Sci U.S.A. 89, 3869-3873.

    Google Scholar 

  • Dix D R, Bridgham J T, Boderius M A, Byersdorfer C A and Eide D J 1994. The FET4 gene encodes the low affinity Fe(II) transport protein of Saccharomyces cerevisiae. J. Biol. Chem. 269, 26092-26099.

    Google Scholar 

  • Eckhardt U and Buckhout T J 1998 Iron assimilation in Chlamydomonas reinhardtii involves ferric reduction and is similar to strategy I higher plants. J. Exp. Bot. 49, 1219-1226.

    Google Scholar 

  • Eide D, Broderius M, Fett J and Guerinot M L 1996 A novel iron-regulated metal transporter from plants identified by functional expression in yeast. Proc. Natl. Acad. Sci. U.S.A. 93, 5624-5628.

    Google Scholar 

  • Fisher M, Gokhman I, Pick U and Zamir A 1997 A structurally novel transferrin-like protein accumulates in the plasma membrane of the unicellular green alga Dunaliella salina grown in high salinities. J. Biol. Chem. 272, 1565-1570.

    Google Scholar 

  • Fisher M, Zamir A and Pick U 1998 Iron uptake by the halotolerant alga Dunaliella is mediated by a plasma membrane transferrin. J Biol. Chem. 273, 17553-17558.

    Google Scholar 

  • Gunshin H, Mackenzie B, Berger U V, Gunshin Y, Romero M F, Boron W F, Nussberger S, Gollan J L and Hediger M A 1997 Cloning and characterization of a mammalian proton-coupled metal-ion transporter. Nature 388, 482-488.

    Google Scholar 

  • Gutierrez J A, Yu J, Rivera S and Wessling-Resnick M 1997 Functional expression, cloning and characterization of SFT, a stimulator of Fe-transport. J. Cell. Biol. 139, 895-905.

    Google Scholar 

  • Harris E H 1988 The Chlamydomonas Sourcebook-A Comprehensive Guide to Biology and Laboratory Use. Academic Press, San Diego.

    Google Scholar 

  • Herbik A, Giritch A, Horstmann C, Becker R, Balzer H J, Baumlein H and Stephan U W 1996 Iron and copper nutrition-dependent changes in protein expression in a tomato wild type and the nicotianamine-free mutant chloronerva. Plant Physiol. 53 533-540.

    Google Scholar 

  • Laemmli U K 1970 Cleavage of structural proteins during the assembly of the head of bacteriophage T4. Nature 227, 680-685.

    Google Scholar 

  • Lannon B and Mason J 1986 The inhibition of bovine ceruloplasmin oxidase activity by thiomolybdates in vivo and in vitro: a reversible interaction. J. Inorg. Biochem. 26, 107-115.

    Google Scholar 

  • Larsson C and Møller I M 1990 The Plant Plasma Membrane. Structure, Function and Molecular Biology. Springer-Verlag, Berlin

    Google Scholar 

  • Lesuisse E, Simon-Casteras M and Labbe P 1998 Siderophoremediated iron uptake in Saccharomyces cerevisiae: the SIT1 gene a ferrioxamine B permease that belongs to the major facilitator superfamily. Microbiology 144, 3455-3462.

    Google Scholar 

  • Lynnes J A, Derzaph T L M and Weger H G 1998 Iron limitation results in induction of ferricyanide reductase and ferric chelate reductase activities in Chlamydomonas reinhardtii. Planta 204, 360-365.

    Google Scholar 

  • Meyer I, Grosset J, Chartier Y and Cleyet-Marel J C 1988 Preparation by two-dimensional electrophoresis of proteins for antibody production: antibody against proteins whose synthesis is reduced by auxin in tobacco mesophyll protoplasts. Electrophoresis 9, 704-712

    Google Scholar 

  • Pinner E, Gruenheid S, Raymond M and Gros P 1997 Functional complementation of the yeast divalent cation transporter family SMF by NRAMP2, a member of the mammalian natural resistance-associated macrophage protein family. J. Biol. Chem. 272, 28933-28938.

    Google Scholar 

  • Randell E W, Parkes J G, Olivieri N F and Templeton D M 1994 Uptake non transferrin-bound iron by both reductive and nonreductive processes is modulated by intracellular iron. J. Biol. Chem. 269, 16046-16053.

    Google Scholar 

  • Riedel H D, Remus A J, Fitscher B A and Stremmel W 1995 Characterization and partial purification of a ferrireductase from human duodenal microvillous membrane. Biochem. J. 309, 745-748.

    Google Scholar 

  • Robinson N J, Procter C M, Connolly E L and Guerinot M L 1999 A ferric-chelate reductase for iron uptake from soils. Nature 397, 694-697.

    Google Scholar 

  • Römheld V and Marschner H 1994 Strategies of plants for acquisition of iron. Plant Soil 165, 261-274.

    Google Scholar 

  • Sadka A, Himmelhoch S and Zamir A 1991 A 150 kilodalton cell surface protein is induced by salt in the halotolerant green alga Dunaliella sauna. Plant Physiol. 95, 822-831.

    Google Scholar 

  • Schmidt A and Buckhout T J 1998 The response of tomato roots (Lycopersicon esculentum Mill) to iron deficiency stress: alterations in the pattern of protein synthesis. J. Exp. Bot. 48, 1909-1918.

    Google Scholar 

  • Stearman R, Yuan D S, Yamaguchi-Iwai Y, Klausner R D and Dancis A 1996 A permease-oxidase complex involved in high affinity iron uptake in yeast. Science 271, 1552-1557.

    Google Scholar 

  • Stephan U W, Schmidtke I and Pich A 1995 Phloem translocation of Fe, Cu, Mn and Zn in Ricinus seedlings in relation to the concentration of nicotianamine, an endogenous chelator of divalent metal ions, in different seedling parts. Plant Soil 165, 181-188.

    Google Scholar 

  • Suzuki K, Itai R, Suzuki K, Nakanishi H, Nishizawa N K, Yoshimura E and Mori S 1998 Formate dehydrogenase, an enzyme of anaerobic metabolism, is induced by iron deficiency in barley roots. Plant Physiol. 116, 725-732.

    Google Scholar 

  • Thomine S, Wang R, Ward, J M, Crawford N M and Schroeder J I 2000 Cadmium and iron transport by members of a plant metal transporter family in Arabidosis with homology to Nramp genes. 97, 4991-4996.

    Google Scholar 

  • Vulpe C D, Kuo Y M, Murphy T L, Cowley L, Askwith C, Libina N, Gitschier J and Anderson G J 1999 Hephaestin, a ceruloplasmin homologue implicated in intestinal iron transport, is defective in the sla mouse. Nat. Genet. 21, 195-199.

    Google Scholar 

  • Wessling-Resnick M 1999 Biochemistry of iron uptake. Crit. Rev. Biochem. Mol. Biol. 34, 285-314.

    Google Scholar 

  • Yamaguchi-Iwai Y, Dancis A and Klausner R D 1995 AFT1: a mediator of iron regulated transcriptional control in Saccharomyces cerevisiae. EMBO J. 14, 1231-1239.

    Google Scholar 

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Herbik, A., Haebel, S. & Buckhout, T.J. Is a ferroxidase involved in the high-affinity iron uptake in Chlamydomonas reinhardtii? . Plant and Soil 241, 1–10 (2002). https://doi.org/10.1023/A:1016037906516

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