Skip to main content
SpringerLink
Log in

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

  • Published:
Plant Molecular Biology Aims and scope Submit manuscript

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.

This is a preview of subscription content, log in via an institution to check access.

Access this article

Log in via an institution

Price excludes VAT (USA)
Tax calculation will be finalised during checkout.

Instant access to the full article PDF.

Institutional subscriptions

Fig. 1
Fig. 2
Fig. 3
Fig. 4
Fig. 5
Fig. 6
Fig. 7
Fig. 8

Similar content being viewed by others

References

  • 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 99:12108–12113

    Article  PubMed  CAS  Google Scholar 

  • 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 415:146–154

    Article  PubMed  CAS  Google Scholar 

  • 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 40:188–199

    Article  PubMed  CAS  Google Scholar 

  • 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 159:1765–1778

    PubMed  CAS  Google Scholar 

  • 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 129:1581–1591

    Article  PubMed  CAS  Google Scholar 

  • 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 62:683–695

    Article  PubMed  CAS  Google Scholar 

  • Chiu W, Niwa Y, Zeng W, Hirano T, Kobayashi H, Sheen J (1996) Engineered GFP as a vital reporter in plants. Curr Biol 6:325–330

    Article  PubMed  CAS  Google Scholar 

  • 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 38:460–472

    Article  PubMed  CAS  Google Scholar 

  • 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 124:95–103

    Article  PubMed  CAS  Google Scholar 

  • 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 276:22901–22909

    Article  PubMed  CAS  Google Scholar 

  • 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 17:628–643

    Article  PubMed  CAS  Google Scholar 

  • 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 135:471–482

    Article  CAS  Google Scholar 

  • 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 98:14837–14842

    Article  PubMed  CAS  Google Scholar 

  • 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 19:635–643

    Article  PubMed  CAS  Google Scholar 

  • 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 97:2486–2490

    Article  PubMed  CAS  Google Scholar 

  • 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 95:13965–13970

    Article  PubMed  CAS  Google Scholar 

  • Hsieh MH, Goodman HM (2005) The Arabidopsis IspH homolog is involved in the plastid nonmevalonate pathway of isoprenoid biosynthesis. Plant Physiol 138:641–653

    Article  PubMed  CAS  Google Scholar 

  • Hsieh MH, Goodman HM (2006) Functional evidence for the involvement of Arabidopsis IspF homolog in the nonmevalonate pathway of plastid isoprenoid biosynthesis. Planta 223:779–784

    Article  PubMed  CAS  Google Scholar 

  • Jackson AO, Larkins BA (1976) Influence of ionic strength, pH, and chelation of divalent metals on isolation of polyribosomes from tobacco leaves. Plant Physiol 57:5–10

    Article  PubMed  CAS  Google Scholar 

  • 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 67:1435–1441

    Article  PubMed  CAS  Google Scholar 

  • 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 25:829–885

    Article  PubMed  CAS  Google Scholar 

  • 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

    Article  PubMed  CAS  Google Scholar 

  • 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 100:6866–6871

    Article  PubMed  CAS  Google Scholar 

  • Leister D (2005) Genomics-based dissection of the cross-talk of chloroplasts with the nucleus and mitochondria in Arabidopsis. Gene 354:110–116

    Article  PubMed  CAS  Google Scholar 

  • Leon P, Arroyo A, Mackenzie S (1998) Nuclear control of plastid and mitochondrial development in higher plants. Annu Rev Plant Physiol Plant Mol Biol 49:453–480

    Article  PubMed  CAS  Google Scholar 

  • Lichtenthaler HK (1999) The 1-deoxy-d-xylulose-5-phosphate pathway of isoprenoid biosynthesis in plants. Annu Rev Plant Physiol Plant Mol Biol 50:47–65

    Article  PubMed  CAS  Google Scholar 

  • Lichtenthaler HK, Wellburn AR (1983) Determination of total carotenoids and chlorophylls a and b of leaf extracts in different solvents. Biochem Soc Trans 11:591–592

    CAS  Google Scholar 

  • 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 95:2105–2110

    Article  PubMed  CAS  Google Scholar 

  • 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 22:503–513

    Article  PubMed  CAS  Google Scholar 

  • 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 97:1062–1067

    Article  PubMed  CAS  Google Scholar 

  • 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 9:649–658

    Article  PubMed  CAS  Google Scholar 

  • 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 98:10499–10504

    Article  PubMed  CAS  Google Scholar 

  • Myerson D (1991) Producing single-stranded DNA probes with the Taq DNA polymerase: A high yield protocol. Biotechniques 10:35–38

    PubMed  Google Scholar 

  • Nott A, Jung HS, Koussevitzky S, Chory J (2006) Plastid-to-nucleus retrograde signaling. Annu Rev Plant Biol 57:739–759

    Article  PubMed  CAS  Google Scholar 

  • 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 215:339–344

    Article  PubMed  CAS  Google Scholar 

  • 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 514:343–346

    Article  PubMed  CAS  Google Scholar 

  • Raghavendra AS, Padmasree K (2003) Beneficial interactions of mitochondrial metabolism with photosynthetic carbon assimilation. Trends Plant Sci 11:546–553

    Article  CAS  Google Scholar 

  • 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 130:1079–1089

    Article  PubMed  CAS  Google Scholar 

  • 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 96:11758–11763

    Article  PubMed  CAS  Google Scholar 

  • 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 99:1158–1163

    Article  PubMed  CAS  Google Scholar 

  • 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 100:1586–1591

    Article  PubMed  CAS  Google Scholar 

  • 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 295:517–524

    PubMed  CAS  Google Scholar 

  • 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 94:12857–12862

    Article  PubMed  CAS  Google Scholar 

  • 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 37:750–761

    Article  PubMed  CAS  Google Scholar 

  • 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 95:9879–9884

    Article  PubMed  CAS  Google Scholar 

  • 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 102:13687–13692

    Article  PubMed  CAS  Google Scholar 

  • 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 144:915–925

    Article  PubMed  CAS  Google Scholar 

  • White RH (1978) Stable isotope studies on the biosynthesis of the thiazole moiety of thiamin in Escherichia coli. Biochemistry 17:3833–3840

    Article  PubMed  CAS  Google Scholar 

Download references

Acknowledgments

We thank Dr. Jen Sheen for providing the GFP expression vector, Mei-Jane Fang for assistance in confocal microscopy, and the Arabidopsis Biological Resource Center for distributing the seeds. This work was supported by grants to M-H H from National Science Council and Academia Sinica of Taiwan.

Author information

Authors and Affiliations

  1. Institute of Plant and Microbial Biology, Academia Sinica, Taipei, 11529, Taiwan

    Ming-Hsiun Hsieh, Chiung-Yun Chang, Shih-Jui Hsu & Ju-Jiun Chen

Authors
  1. Ming-Hsiun Hsieh
    View author publications

    You can also search for this author in PubMed Google Scholar

  2. Chiung-Yun Chang
    View author publications

    You can also search for this author in PubMed Google Scholar

  3. Shih-Jui Hsu
    View author publications

    You can also search for this author in PubMed Google Scholar

  4. Ju-Jiun Chen
    View author publications

    You can also search for this author in PubMed Google Scholar

Corresponding author

Correspondence to Ming-Hsiun Hsieh.

Electronic supplementary material

Below is the link to the electronic supplementary material.

(DOC 28 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)

Rights and permissions

Reprints and permissions

About this article

Cite this article

Hsieh, MH., Chang, CY., Hsu, SJ. et al. Chloroplast localization of methylerythritol 4-phosphate pathway enzymes and regulation of mitochondrial genes in ispD and ispE albino mutants in Arabidopsis. Plant Mol Biol 66, 663–673 (2008). https://doi.org/10.1007/s11103-008-9297-5

Download citation

  • Received:

  • Accepted:

  • Published:

  • Issue Date:

  • DOI: https://doi.org/10.1007/s11103-008-9297-5

Keywords

Search

Navigation