Plant Molecular Biology

, Volume 66, Issue 6, pp 663–673

Chloroplast localization of methylerythritol 4-phosphate pathway enzymes and regulation of mitochondrial genes in ispD and ispE albino mutants in Arabidopsis

  • Ming-Hsiun Hsieh
  • Chiung-Yun Chang
  • Shih-Jui Hsu
  • Ju-Jiun Chen
Article

Abstract

Plant isoprenoids are derived from two independent pathways, the cytosolic mevalonate pathway and the plastid methylerythritol 4-phosphate (MEP) pathway. We used green fluorescent fusion protein assays to demonstrate that the Arabidopsis MEP pathway enzymes are localized to the chloroplast. We have also characterized three Arabidopsis albino mutants, ispD-1, ispD-2 and ispE-1, which have T-DNA insertions in the IspD and IspE genes of the MEP pathway. Levels of photosynthetic pigments are almost undetectable in these albino mutants. Instead of thylakoids, the ispD and ispE mutant chloroplasts are filled with large vesicles. Impairments in chloroplast development and functions may signal changes in the expression of nuclear, chloroplast and mitochondrial genes. We used northern blot analysis to examine the expression of photosynthetic and respiratory genes in the ispD and ispE albino mutants. Steady-state mRNA levels of nucleus- and chloroplast-encoded photosynthetic genes are significantly decreased in the albino mutants. In contrast, transcript levels of nuclear and mitochondrial genes encoding subunits of the mitochondrial electron transport chain are increased or not affected in these mutants. Genomic Southern blot analysis revealed that the DNA amounts of mitochondrial genes are not enhanced in the ispD and ispE albino mutants. These results support the notion that the functional state of chloroplasts may affect the expression of nuclear and mitochondrial genes. The up-regulation of mitochondrial genes in the albino mutants is not caused by changes of mitochondrial DNA copy number in Arabidopsis.

Keywords

Arabidopsis Albino Chloroplast Isoprenoid biosynthesis Methylerythritol 4-phosphate pathway Mitochondrial gene expression 

Supplementary material

11103_2008_9297_MOESM2_ESM.tif (685 kb)
Supplementary Figure 1 Verification of T-DNA insertion lines by genomic Southern blot analysis. (a) Ten micrograms of genomic DNA extracted from wild type (WT), albino segregants of ispD-1 (SALK_042163) and ispD-2 (SALK_030640) T-DNA lines were digested with Bam HI and subjected to Southern blot analysis. (b) Ten micrograms of genomic DNA extracted from wild type and ispE-1 (SALK_107310) albino plants were digested with Eco RV and subjected to Southern blot analysis. The wild type IspD and IspE alleles were not detected in ispD-1, ispD-2 and ispE-1 T-DNA mutants, respectively. Primers for making 32P-labeled IspD probe and DIG-labeled IspE probe are listed in Supplementary Table 1. (TIF 685 kb)

References

  1. Adam P, Hecht S, Eisenreich W, Kaiser J, Grawert T, Arigoni D, Bacher A, Rohdich F (2002) Biosynthesis of terpenes: studies on 1-hydroxy-2-methyl-2-(E)-butenyl 4-diphosphate reductase. Proc Natl Acad Sci USA 99:12108–12113PubMedCrossRefGoogle Scholar
  2. Bick JA, Lange BM (2003) Metabolic cross talk between cytosolic and plastidial pathways of isoprenoid biosynthesis: unidirectional transport of intermediates across the chloroplast envelope membrane. Arch Biochem Biophys 415:146–154PubMedCrossRefGoogle Scholar
  3. Botella-Pavía P, Besumbes O, Phillips MA, Carretero-Paulet L, Boronat A, Rodríguez-Concepción M (2004) Regulation of carotenoid biosynthesis in plants: evidence for a key role of hydroxymethylbutenyl diphosphate reductase in controlling the supply of plastidial isoprenoid precursors. Plant J 40:188–199PubMedCrossRefGoogle Scholar
  4. Budziszewski GJ, Lewis SP, Glover LW, Reineke J, Jones G, Ziemnik LS, Lonowski J, Nyfeler B, Aux G, Zhou Q, McElver J, Patton DA, Martienssen R, Grossniklaus U, Ma H, Law M, Levin JZ (2001) Arabidopsis genes essential for seedling viability: isolation of insertional mutants and molecular cloning. Genetics 159:1765–1778PubMedGoogle Scholar
  5. Carretero-Paulet L, Ahumada I, Cunillera N, Rodriguez-Concepcion M, Ferrer A, Boronat A, Campos N (2002) Expression and molecular analysis of the Arabidopsis DXR gene encoding 1-deoxy-d-xylulose 5-phosphate reductoisomerase, the first committed enzyme of the 2-C-methyl-d-erythritol 4-phosphate pathway. Plant Physiol 129:1581–1591PubMedCrossRefGoogle Scholar
  6. Carretero-Paulet L, Cairó A, Botella-Pavía P, Besumbes O, Campos N, Boronat A, Rodríguez-Concepción M (2006) Enhanced flux through the methylerythritol 4-phosphate pathway in Arabidopsis plants overexpressing deoxyxylulose 5-phosphate reductoisomerase. Plant Mol Biol 62:683–695PubMedCrossRefGoogle Scholar
  7. Chiu W, Niwa Y, Zeng W, Hirano T, Kobayashi H, Sheen J (1996) Engineered GFP as a vital reporter in plants. Curr Biol 6:325–330PubMedCrossRefGoogle Scholar
  8. Emanuel C, Weihe A, Graner A, Hess WR, Börner T (2004) Chloroplast development affects expression of phage-type RNA polymerases in barley leaves. Plant J 38:460–472PubMedCrossRefGoogle Scholar
  9. Estevez JM, Cantero A, Romero C, Kawaide H, Jimenez LF, Kuzuyama T, Seto H, Kamiya Y, Leon P (2000) Analysis of the expression of CLA1, a gene that encodes the 1-deoxyxylulose 5-phosphate synthase of the 2-C-methyl-d-erythritol-4-phosphate pathway in Arabidopsis. Plant Physiol 124:95–103PubMedCrossRefGoogle Scholar
  10. Estevez JM, Cantero A, Reindl A, Reichler S, Leon P (2001) 1-Deoxyxylulose 5-phosphate synthase, a limiting enzyme for plastidic isoprenoid biosynthesis in plants. J Biol Chem 276:22901–22909PubMedCrossRefGoogle Scholar
  11. Guevara-Garcia A, San Roman C, Arroyo A, Cortes ME, de la Luz Gutierrez-Nava M, Leon P (2005) Characterization of the Arabidopsis clb6 mutant illustrates the importance of posttranscriptional regulation of the methyl-d-erythritol 4-phosphate pathway. Plant Cell 17:628–643PubMedCrossRefGoogle Scholar
  12. Gutierrez-Nava M de L, Gillmor CS, Jimenez LF, Guevara-Garcia A, Leon P (2004) CHLOROPLAST BIOGENESIS genes act cell and noncell autonomously in early chloroplast development. Plant Physiol 135:471–482CrossRefGoogle Scholar
  13. Hecht S, Eisenreich W, Adam P, Amslinger S, Kis K, Bacher A, Arigoni D, Rohdich F (2001) Studies on the nonmevalonate pathway to terpenes: the role of the GcpE (IspG) protein. Proc Natl Acad Sci USA 98:14837–14842PubMedCrossRefGoogle Scholar
  14. Hedtke B, Wagner I, Borner T, Hess WR (1999) Inter-organellar crosstalk in higher plants: impaired chloroplast development affects mitochondrial gene and transcript levels. Plant J 19:635–643PubMedCrossRefGoogle Scholar
  15. Herz S, Wungsintaweekul J, Schuhr CA, Hecht S, Luttgen H, Sagner S, Fellermeier M, Eisenreich W, Zenk MH, Bacher A, Rohdich F (2000) Biosynthesis of terpenoids: YgbB protein converts 4-diphosphocytidyl-2C-methyl-d-erythritol 2-phosphate to 2C-methyl-d-erythritol 2,4-cyclodiphosphate. Proc Natl Acad Sci USA 97:2486–2490PubMedCrossRefGoogle Scholar
  16. Hsieh MH, Lam HM, van de Loo FJ, Coruzzi G (1998) A PII-like protein in Arabidopsis: putative role in nitrogen sensing. Proc Natl Acad Sci USA 95:13965–13970PubMedCrossRefGoogle Scholar
  17. Hsieh MH, Goodman HM (2005) The Arabidopsis IspH homolog is involved in the plastid nonmevalonate pathway of isoprenoid biosynthesis. Plant Physiol 138:641–653PubMedCrossRefGoogle Scholar
  18. Hsieh MH, Goodman HM (2006) Functional evidence for the involvement of Arabidopsis IspF homolog in the nonmevalonate pathway of plastid isoprenoid biosynthesis. Planta 223:779–784PubMedCrossRefGoogle Scholar
  19. Jackson AO, Larkins BA (1976) Influence of ionic strength, pH, and chelation of divalent metals on isolation of polyribosomes from tobacco leaves. Plant Physiol 57:5–10PubMedCrossRefGoogle Scholar
  20. Kim SM, Kuzuyama T, Chang YJ, Kwon HJ, Kim SU (2006a) Cloning and functional characterization of 2-C-methyl-d-erythritol 4-phosphate cytidyltransferase (GbMECT) gene from Ginkgo biloba. Phytochemistry 67:1435–1441PubMedCrossRefGoogle Scholar
  21. Kim SM, Kuzuyama T, Chang YJ, Kim SU (2006b) Cloning and characterization of 2-C-methyl-D: -erythritol 2,4-cyclodiphosphate synthase (MECS) gene from Ginkgo biloba. Plant Cell Rep 25:829–885PubMedCrossRefGoogle Scholar
  22. Koussevitzky S, Nott A, Mockler TC, Hong F, Sachetto-Martins G, Surpin M, Lim J, Mittler R, Chory J (2007) Signals from chloroplasts converge to regulate nuclear gene expression. Science 316:715–719 PubMedCrossRefGoogle Scholar
  23. Laule O, Furholz A, Chang HS, Zhu T, Wang X, Heifetz PB, Gruissem W, Lange M (2003) Crosstalk between cytosolic and plastidial pathways of isoprenoid biosynthesis in Arabidopsis thaliana. Proc Natl Acad Sci USA 100:6866–6871PubMedCrossRefGoogle Scholar
  24. Leister D (2005) Genomics-based dissection of the cross-talk of chloroplasts with the nucleus and mitochondria in Arabidopsis. Gene 354:110–116PubMedCrossRefGoogle Scholar
  25. Leon P, Arroyo A, Mackenzie S (1998) Nuclear control of plastid and mitochondrial development in higher plants. Annu Rev Plant Physiol Plant Mol Biol 49:453–480PubMedCrossRefGoogle Scholar
  26. Lichtenthaler HK (1999) The 1-deoxy-d-xylulose-5-phosphate pathway of isoprenoid biosynthesis in plants. Annu Rev Plant Physiol Plant Mol Biol 50:47–65PubMedCrossRefGoogle Scholar
  27. Lichtenthaler HK, Wellburn AR (1983) Determination of total carotenoids and chlorophylls a and b of leaf extracts in different solvents. Biochem Soc Trans 11:591–592Google Scholar
  28. Lois LM, Campos N, Putra SR, Danielsen K, Rohmer M, Boronat A (1998) Cloning and characterization of a gene from Escherichia coli encoding a transketolase-like enzyme that catalyzes the synthesis of d-1-deoxyxylulose 5-phosphate, a common precursor for isoprenoid, thiamin, and pyridoxol biosynthesis. Proc Natl Acad Sci USA 95:2105–2110PubMedCrossRefGoogle Scholar
  29. Lois LM, Rodriguez-Concepcion M, Gallego F, Campos N, Boronat A (2000) Carotenoid biosynthesis during tomato fruit development: regulatory role of 1-deoxy-d-xylulose 5-phosphate synthase. Plant J 22:503–513PubMedCrossRefGoogle Scholar
  30. Luttgen H, Rohdich F, Herz S, Wungsintaweekul J, Hecht S, Schuhr CA, Fellermeier M, Sagner S, Zenk MH, Bacher A, Eisenreich W (2000) Biosynthesis of terpenoids: YchB protein of Escherichia coli phosphorylates the 2-hydroxy group of 4-diphosphocytidyl-2C-methyl-d-erythritol. Proc Natl Acad Sci USA 97:1062–1067PubMedCrossRefGoogle Scholar
  31. Mandel MA, Feldmann KA, Herrera-Estrella L, Rocha-Sosa M, Leon P (1996) CLA1, a novel gene required for chloroplast development, is highly conserved in evolution. Plant J 9:649–658PubMedCrossRefGoogle Scholar
  32. Motohashi R, Nagata N, Ito T, Takahashi S, Hobo T, Yoshida S, Shinozaki K (2001) An essential role of a TatC homologue of a pH- dependent protein transporter in thylakoid membrane formation during chloroplast development in Arabidopsis thaliana. Proc Natl Acad Sci USA 98:10499–10504PubMedCrossRefGoogle Scholar
  33. Myerson D (1991) Producing single-stranded DNA probes with the Taq DNA polymerase: A high yield protocol. Biotechniques 10:35–38PubMedGoogle Scholar
  34. Nott A, Jung HS, Koussevitzky S, Chory J (2006) Plastid-to-nucleus retrograde signaling. Annu Rev Plant Biol 57:739–759PubMedCrossRefGoogle Scholar
  35. Okada K, Kawaide H, Kuzuyama T, Seto H, Curtis IS, Kamiya Y (2002) Antisense and chemical suppression of the nonmevalonate pathway affects ent-kaurene biosynthesis in Arabidopsis. Planta 215:339–344PubMedCrossRefGoogle Scholar
  36. Querol J, Campos N, Imperial S, Boronat A, Rodriguez-Concepcion M (2002) Functional analysis of the Arabidopsis thaliana GCPE protein involved in plastid isoprenoid biosynthesis. FEBS Lett 514:343–346PubMedCrossRefGoogle Scholar
  37. Raghavendra AS, Padmasree K (2003) Beneficial interactions of mitochondrial metabolism with photosynthetic carbon assimilation. Trends Plant Sci 11:546–553CrossRefGoogle Scholar
  38. Rodriguez-Concepcion M, Boronat A (2002) Elucidation of the methylerythritol phosphate pathway for isoprenoid biosynthesis in bacteria and plastids. A metabolic milestone achieved through genomics. Plant Physiol 130:1079–1089PubMedCrossRefGoogle Scholar
  39. Rohdich F, Wungsintaweekul J, Fellermeier M, Sagner S, Herz S, Kis K, Eisenreich W, Bacher A, Zenk MH (1999) Cytidine 5′-triphosphate-dependent biosynthesis of isoprenoids: YgbP protein of Escherichia coli catalyzes the formation of 4-diphosphocytidyl-2-C-methylerythritol. Proc Natl Acad Sci USA 96:11758–11763PubMedCrossRefGoogle Scholar
  40. Rohdich F, Hecht S, Gartner K, Adam P, Krieger C, Amslinger S, Arigoni D, Bacher A, Eisenreich W (2002) Studies on the nonmevalonate terpene biosynthetic pathway: metabolic role of IspH (LytB) protein. Proc Natl Acad Sci USA 99:1158–1163PubMedCrossRefGoogle Scholar
  41. Rohdich F, Zepeck F, Adam P, Hecht S, Kaiser J, Laupitz R, Grawert T, Amslinger S, Eisenreich W, Bacher A, Arigoni D (2003) The deoxyxylulose phosphate pathway of isoprenoid biosynthesis: studies on the mechanisms of the reactions catalyzed by IspG and IspH protein. Proc Natl Acad Sci USA 100:1586–1591PubMedCrossRefGoogle Scholar
  42. Rohmer M, Knani M, Simonin P, Sutter B, Sahm H (1993) Isoprenoid biosynthesis in bacteria: a novel pathway for the early steps leading to isopentenyl diphosphate. Biochem J 295:517–524PubMedGoogle Scholar
  43. Sprenger GA, Schorken U, Wiegert T, Grolle S, de Graaf AA, Taylor SV, Begley TP, Bringer-Meyer S, Sahm H (1997) Identification of a thiamin-dependent synthase in Escherichia coli required for the formation of the 1-deoxy-d-xylulose 5-phosphate precursor to isoprenoids, thiamin, and pyridoxol. Proc Natl Acad Sci USA 94:12857–12862PubMedCrossRefGoogle Scholar
  44. Suzuki M, Kamide Y, Nagata N, Seki H, Ohyama K, Kato H, Masuda K, Sato S, Kato T, Tabata S, Yoshida S, Muranaka T (2004) Loss of function of 3-hydroxy-3-methylglutaryl coenzyme A reductase 1 (HMG1) in Arabidopsis leads to dwarfing, early senescence and male sterility, and reduced sterol levels. Plant J 37:750–761PubMedCrossRefGoogle Scholar
  45. Takahashi S, Kuzuyama T, Watanabe H, Seto H (1998) A 1-deoxy-d-xylulose 5-phosphate reductoisomerase catalyzing the formation of 2-C-methyl-d-erythritol 4-phosphate in an alternative nonmevalonate pathway for terpenoid biosynthesis. Proc Natl Acad Sci USA 95:9879–9884PubMedCrossRefGoogle Scholar
  46. Tambasco-Studart M, Titiz O, Raschle T, Forster G, Amrhein N, Fitzpatrick TB (2005) Vitamin B6 biosynthesis in higher plants. Proc Natl Acad Sci USA 102:13687–13692PubMedCrossRefGoogle Scholar
  47. Tambasco-Studart M, Tews I, Amrhein N, Fitzpatrick TB (2007) Functional analysis of PDX2 from Arabidopsis, a glutaminase involved in vitamin B6 biosynthesis. Plant Physiol 144:915–925PubMedCrossRefGoogle Scholar
  48. White RH (1978) Stable isotope studies on the biosynthesis of the thiazole moiety of thiamin in Escherichia coli. Biochemistry 17:3833–3840PubMedCrossRefGoogle Scholar

Copyright information

© Springer Science+Business Media B.V. 2008

Authors and Affiliations

  • Ming-Hsiun Hsieh
    • 1
  • Chiung-Yun Chang
    • 1
  • Shih-Jui Hsu
    • 1
  • Ju-Jiun Chen
    • 1
  1. 1.Institute of Plant and Microbial BiologyAcademia SinicaTaipeiTaiwan

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