3-hydroxy-3-methyl glutaryl coenzyme A reductase

An essential actor in the biosynthesis of cantharidin in the blister beetle Epicauta chinensis Laporte

S. Lü, M. Jiang, T. Huo, Xianchun Li, Y. Zhang

Research output: Contribution to journalArticle

8 Citations (Scopus)

Abstract

Cantharidin (C10H12O4) is a monoterpene defensive toxin in insects involved in chemical defence as well as in courtship and mating behaviours. It is relatively well known in the medical literature because of its high anticancer activity and as an effective therapy for molluscum contagiosum. However, little is known about its biosynthesis pathway in vivo, and no enzyme involved in cantharidin biosynthesis has been identified. The purpose of this study was to identify the crucial enzyme that is involved in the biosynthesis of cantharidin. Using the homology cloning method, a 3-hydroxy-3-methyl glutaryl coenzyme A reductase (HMGR) gene, the rate-limiting enzyme in the mevalonate pathway, was cloned from the blister beetle Epicauta chinensis. Quantitative reverse transcription PCR and gas chromatography methods revealed that the HMGR transcripts had a positive correlation with cantharidin production in the beetles (R = 0.891). RNA interference (RNAi) knockdown of HMGR mRNA expression was achieved by microinjection of a specific double-stranded RNA with more than 90% RNAi efficiency, and an apparent decrease of cantharidin production was observed. Furthermore, the HMGR mRNA was greatly upregulated by exogenous juvenile hormone III (JH III), and cantharidin production was also raised in males; however, when injecting the JH III with RNAi of HMGR mRNA at the same time, cantharidin production did not rise. These results demonstrate that HMGR is an essential enzyme in cantharidin biosynthesis in the blister beetle E. chinensis, which further verifies previous research results demonstrating that cantharidin is synthesized de novo by the mevalonate pathway in blister beetles.

Original languageEnglish (US)
Pages (from-to)58-71
Number of pages14
JournalInsect Molecular Biology
Volume25
Issue number1
DOIs
StatePublished - Feb 1 2016

Fingerprint

Epicauta
Cantharidin
cantharidin
Meloidae
coenzyme A
Beetles
Blister
Oxidoreductases
biosynthesis
RNA Interference
RNA interference
Mevalonic Acid
juvenile hormones
Enzymes
enzymes
Messenger RNA
Molluscum Contagiosum
Courtship
glutaryl-coenzyme A
Monoterpenes

Keywords

  • 3-hydroxy-3-methyl glutaryl coenzyme A reductase (HMGR)
  • cantharidin
  • Epicauta chinensis
  • juvenile hormone III (JH III)
  • mevalonate pathway

ASJC Scopus subject areas

  • Insect Science
  • Molecular Biology
  • Genetics

Cite this

3-hydroxy-3-methyl glutaryl coenzyme A reductase : An essential actor in the biosynthesis of cantharidin in the blister beetle Epicauta chinensis Laporte. / Lü, S.; Jiang, M.; Huo, T.; Li, Xianchun; Zhang, Y.

In: Insect Molecular Biology, Vol. 25, No. 1, 01.02.2016, p. 58-71.

Research output: Contribution to journalArticle

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abstract = "Cantharidin (C10H12O4) is a monoterpene defensive toxin in insects involved in chemical defence as well as in courtship and mating behaviours. It is relatively well known in the medical literature because of its high anticancer activity and as an effective therapy for molluscum contagiosum. However, little is known about its biosynthesis pathway in vivo, and no enzyme involved in cantharidin biosynthesis has been identified. The purpose of this study was to identify the crucial enzyme that is involved in the biosynthesis of cantharidin. Using the homology cloning method, a 3-hydroxy-3-methyl glutaryl coenzyme A reductase (HMGR) gene, the rate-limiting enzyme in the mevalonate pathway, was cloned from the blister beetle Epicauta chinensis. Quantitative reverse transcription PCR and gas chromatography methods revealed that the HMGR transcripts had a positive correlation with cantharidin production in the beetles (R = 0.891). RNA interference (RNAi) knockdown of HMGR mRNA expression was achieved by microinjection of a specific double-stranded RNA with more than 90{\%} RNAi efficiency, and an apparent decrease of cantharidin production was observed. Furthermore, the HMGR mRNA was greatly upregulated by exogenous juvenile hormone III (JH III), and cantharidin production was also raised in males; however, when injecting the JH III with RNAi of HMGR mRNA at the same time, cantharidin production did not rise. These results demonstrate that HMGR is an essential enzyme in cantharidin biosynthesis in the blister beetle E. chinensis, which further verifies previous research results demonstrating that cantharidin is synthesized de novo by the mevalonate pathway in blister beetles.",
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