Publication:
Cloning and expression of 1-deoxy-d-xylulose 5-phosphate synthase cDNA from Croton stellatopilosus and expression of 2C-methyl-d-erythritol 4-phosphate synthase and geranylgeranyl diphosphate synthase, key enzymes of plaunotol biosynthesis

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Issued Date

2010

Resource Type

Article

File Type

application/pdf

ISSN

1761617

DOI

10.1016/j.jplph.2009.09.001

Other identifier(s)

2-s2.0-75149166956

Rights Holder(s)

Scopus

Bibliographic Citation

Journal of Plant Physiology. Vol 167, No.4 (2010), p.292-300

Suggested Citation

Sitthithaworn W., Wungsintaweekul J., Sirisuntipong T., Charoonratana T., Ebizuka Y., De-Eknamkul W. Cloning and expression of 1-deoxy-d-xylulose 5-phosphate synthase cDNA from Croton stellatopilosus and expression of 2C-methyl-d-erythritol 4-phosphate synthase and geranylgeranyl diphosphate synthase, key enzymes of plaunotol biosynthesis. Journal of Plant Physiology. Vol 167, No.4 (2010), p.292-300. doi:10.1016/j.jplph.2009.09.001 Retrieved from: https://hdl.handle.net/20.500.14740/7604

Title

Cloning and expression of 1-deoxy-d-xylulose 5-phosphate synthase cDNA from Croton stellatopilosus and expression of 2C-methyl-d-erythritol 4-phosphate synthase and geranylgeranyl diphosphate synthase, key enzymes of plaunotol biosynthesis

Author(s)

Sitthithaworn W.
 Wungsintaweekul J.
 Sirisuntipong T.
 Charoonratana T.
 Ebizuka Y.
 De-Eknamkul W.

Abstract

1-Deoxy-d-xylulose 5-phosphate synthase (DXS, EC: 4.1.3.37), the first enzyme in the 2C-methyl-d-erythritol 4-phosphate (MEP) pathway, is known to be responsible for the rate-limiting step of isoprenoid biosynthesis in Escherichia coli and Arabidopsis thaliana. In this study, the dxs gene from Croton stellatopilosus, designated csdxs, was cloned from leaf tissue using the rapid amplification of cDNA ends (RACE) technique. Leaves of C. stellatopilosus contain plaunotol, an acyclic diterpene alcohol. The csdxs cDNA containing the open reading frame of 2163 base pairs appeared to encode a polypeptide of 720 amino acids. Analysis of the deduced amino acid sequence revealed that the NH2-terminus of CSDXS carried a chloroplast transit peptide, a thiamine diphosphate binding site, and a transketolase motif, which are the important characteristics of DXS enzymes in higher plants. Multiple alignments of CSDXS with other plant DXSs have indicated that CSDXS has identity ranging between 68% and 89%. Expression levels of csdxs and genes encoding key enzymes in the plaunotol biosynthetic pathway, namely 2C-methyl-d-erythritol 4-phosphate synthase (meps) and geranylgeranyl diphosphate synthase (ggpps), were analysed by measuring transcript levels in leaves of different developmental stages. The results showed that dxs, meps, and ggpps are all active in young leaves prior to full expansion when plaunotol is synthesised from the DXP precursor in chloroplasts. The dense presence of chloroplasts and oil globules in the palisade cells of these leaves support the view that these genes are involved in plaunotol biosynthesis in chloroplast-containing tissues. Crown Copyright © 2009.

Subject(s)

1 deoxy D xylulose 5 phosphate reductoisomerase
 1-deoxy-D-xylulose 5-phosphate reductoisomerase
 Complementary DNA
 Deoxyxylulose 5 phosphate synthase
 Deoxyxylulose-5-phosphate synthase
 Farnesyl trans transferase
 Fatty alcohol
 Isomerase
 Messenger RNA
 Multienzyme complex
 Oxidoreductase
 Plaunotol
 Transferase
 Amino acid sequence
 Antibody specificity
 Article
 Chemistry
 Croton
 Enzymology
 Gene expression profiling
 Gene expression regulation
 Genetics
 Metabolism
 Molecular cloning
 Molecular genetics
 Nucleotide sequence
 Phylogeny
 Plant
 Plant leaf
 Sequence alignment
 Ultrastructure
 Aldose-Ketose Isomerases
 Amino Acid Sequence
 Cloning, Molecular
 Croton
 DNA, Complementary
 Farnesyltranstransferase
 Fatty Alcohols
 Gene Expression Profiling
 Gene Expression Regulation, Enzymologic
 Gene Expression Regulation, Plant
 Molecular Sequence Data
 Multienzyme Complexes
 Organ Specificity
 Oxidoreductases
 Phylogeny
 Plant Leaves
 Plant Shoots
 RNA, Messenger
 Sequence Alignment
 Transferases
 Arabidopsis
 Arabidopsis thaliana
 Embryophyta
 Escherichia coli

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URI

https://hdl.handle.net/20.500.14740/7604

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Scopus: Year 1983-2021

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