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Functional evidence for the involvement of Arabidopsis IspF homolog in the nonmevalonate pathway of plastid isoprenoid biosynthesis

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Abstract

There are two independent pathways, the cytosolic mevalonate (MVA) pathway and the plastid nonmevalonate (nonMVA) pathway, to synthesize isopentenyl diphosphate and dimethylallyl diphosphate in plants. Carotenoids and the phytyl side chain of chlorophylls are isoprenoids derived from the plastid nonMVA pathway. All enzymes involved in the nonMVA pathway have been identified in Escherichia coli. The E. coli IspF protein catalyzes a unique cyclization reaction to convert 4-diphosphocytidyl-2-C-methyl-D-erythritol 2-phosphate into 2-C-methyl-D-erythritol 2,4-cyclodiphosphate in the nonMVA pathway. We have characterized an Arabidopsis T-DNA insertion mutant, ispF-1, which has a null mutation in the IspF gene. Homozygous ispF-1 mutants are albino lethal and the IspF transcripts are undetectable in these plants. Moreover, the ispF-1 mutant chloroplasts are filled with vesicles instead of thylakoids. Amino acid sequence alignment reveals that the IspF proteins are highly conserved between plants and bacteria. Interestingly, expression of the Arabidopsis IspF protein can rescue the lethal phenotype of an E. coli ispF mutant. These results indicate that the Arabidopsis IspF may share similar enzymatic mechanisms with the E. coli protein.

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Abbreviations

MVA:

Mevalonate

nonMVA:

Nonmevalonate

IPP:

Isopentenyl diphosphate

DMAPP:

Dimethylallyl diphosphate

DXP:

1-Deoxy-d-xylulose-5-phosphate

DXS:

DXP synthase

DXR (IspC):

DXP reductoisomerase

MEP:

2-C-Methyl-D-erythritol 4-phosphate

CDP-ME:

4-Diphosphocytidyl-2-C-methyl-D-erythritol

CMS (IspD):

CDP-ME synthase

CMK (IspE):

CDP-ME kinase

CDP-MEP:

4-Diphosphocytidyl-2-C-methyl-D-erythritol 2-phosphate

ME-cPP:

2-C-Methyl-D-erythritol 2,4-cyclodiphosphate

MCS (IspF):

ME-cPP synthase

HMBPP:

1-Hydroxy-2-methyl-2-(E)-butenyl 4-diphosphate

HDS (IspG):

HMBPP synthase

HDR (IspH):

HMBPP reductase

MS medium:

Murashige and Skoog medium

DIG:

Digoxigenin

IPTG:

Isopropyl-β-D-thiogalactopyranoside

GGPP:

Geranylgeranyl diphosphate

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Acknowledgements

We thank Dr. E. Brown for the E. coli ispF mutant strain and the Arabidopsis Biological Resource Center for providing the ispF-1 mutant seeds.

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Authors and Affiliations

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

    Ming-Hsiun Hsieh

  2. Department of Genetics, Harvard Medical School and Department of Molecular Biology, Massachusetts General Hospital, Boston, MA, 02114, USA

    Howard M. Goodman

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Correspondence to Ming-Hsiun Hsieh.

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Hsieh, MH., Goodman, H.M. Functional evidence for the involvement of Arabidopsis IspF homolog in the nonmevalonate pathway of plastid isoprenoid biosynthesis. Planta 223, 779–784 (2006). https://doi.org/10.1007/s00425-005-0140-9

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  • DOI: https://doi.org/10.1007/s00425-005-0140-9

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