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Two copies of 4-(cytidine 5′-diphospho)-2-C-methyl-d-erythritol kinase (CMK) gene in Ginkgo biloba: molecular cloning and functional characterization

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Abstract

4-(Cytidine 5′-diphospho)-2-C-methyl-d-erythritol kinase (CMK or YchB), the fourth enzyme of the 2-C-methyl-d-erythritol 4-phosphate pathway, phosphorylates the 2-hydroxyl group of 4-(cytidine 5′-diphospho)-2-C-methyl-d-erythritol in the presence of ATP. Two isogenes encoding CMK (GbCMK1 and GbCMK2) were cloned and characterized from Ginkgo biloba. The activities of both isozymes were confirmed by complementation assay using Escherichia coli NMW29, a ychB knock-out mutant. The transcript profiles of GbCMKs in the radicles and the cotyledons of the cultured Ginkgo biloba embryos demonstrated that the transcript levels of GbCMK1 were similar in both organs, whereas that of GbCMK2 was predominantly high in the ginkgolide-synthesizing radicles. Selective increases in the transcript abundance of GbCMK2 in the radicles, induced by light and methyl jasmonate treatments, were observed. These differential induction patterns of the transcripts imply GbCMK1 and GbCMK2 respectively have high correlations with the primary and the secondary metabolisms. The transit peptides of both isozymes delivered the fused green fluorescent protein (GFP) into the chloroplast in the Arabidopsis and the Nicotiana transient expression systems; interestingly, the transit peptide of GbCMK1 delivered the GFP protein into the cytosol and the nucleus in addition to the chloroplasts.

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Abbreviations

CDP-ME:

4-(Cytidine 5′-diphospho)-2-C-methyl-d-erythritol

CMK:

CDP-ME kinase

DMAPP:

Dimethylallyl diphosphate

DXR:

1-Deoxy-d-xylulose 5-phosphate reductoisomerase

DXS:

1-Deoxy-d-xylulose 5-phosphate synthase

GFP:

Green fluorescent protein

GGPP:

Geranylgeranyl diphosphate

HMGR:

HMG-CoA reductase

HMGS:

HMG-CoA synthase

IDS:

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

IPP:

Isopentenyl diphosphate

MEP:

2-C-methyl-d-erythritol 4-phosphate

MVA:

Mevalonic acid

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Acknowledgments

The authors appreciate the supports given by the Korea Science and Engineering Foundation (KOSEF 981-0608-040-2) through PMRC, and the Brain Korea 21 program administered by the Ministry of Education, Science and Technology, Korea, through the Department of Agricultural Biotechnology, SNU.

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Authors and Affiliations

  1. Department of Agricultural Biotechnology, Research Institute for Agriculture and Life Sciences, Seoul National University, San 56-1, Sillim-dong, Gwanak-gu, Seoul, 151-921, South Korea

    Sang-Min Kim, Yeon-Bok Kim & Soo-Un Kim

  2. Laboratory of Cell Biotechnology, Biotechnology Research Center, University of Tokyo, Tokyo, 113-8657, Japan

    Tomohisa Kuzuyama

  3. Plant Metabolism Research Center, Kyung Hee University, Yongin, 449-701, South Korea

    Soo-Un Kim

Authors
  1. Sang-Min Kim
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  2. Yeon-Bok Kim
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  3. Tomohisa Kuzuyama
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  4. Soo-Un Kim
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Corresponding author

Correspondence to Soo-Un Kim.

Additional information

S.-M. Kim and Y.-B. Kim contributed equally to this work.

GenBank accession numbers: GbCMK1 (DQ102358), GbCMK2 (DQ102359).

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Kim, SM., Kim, YB., Kuzuyama, T. et al. Two copies of 4-(cytidine 5′-diphospho)-2-C-methyl-d-erythritol kinase (CMK) gene in Ginkgo biloba: molecular cloning and functional characterization. Planta 228, 941–950 (2008). https://doi.org/10.1007/s00425-008-0794-1

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  • DOI: https://doi.org/10.1007/s00425-008-0794-1

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