Cloning and expressing a highly functional and substrate specific farnesoic acid o-methyltransferase from the Asian citrus psyllid (Diaphorina citri Kuwayama)

Van Ekert, Evelien; Shatters, Jr., Robert G.; Rougé, Pierre; Powell, Charles A.; Smagghe, Guy; Borovsky, Dov

doi:10.1016/j.fob.2015.03.012

Title: Cloning and expressing a highly functional and substrate specific farnesoic acid o-methyltransferase from the Asian citrus psyllid (Diaphorina citri Kuwayama)

Journal Article · 28 March 2015 · FEBS Open Bio

DOI: https://doi.org/10.1016/j.fob.2015.03.012 · OSTI ID:1209195

Van Ekert, Evelien ^[1]; Shatters, Jr., Robert G. ^[1]; Rougé, Pierre ^[2]; Powell, Charles A. ^[3]; Smagghe, Guy ^[4]; Borovsky, Dov ^[1]

US Dept. of Agriculture (USDA), Ft. Pierce, FL (United States). Agricultural Research Service
Univ. de Toulouse (France)
Univ. of Florida, Fort Pierce, FL (United States). Indian River Research and Education Center
Univ. Gent (Belgium)

The Asian citrus psyllid, Diaphorina citri , transmits a phloem-limited bacterium, Candidatus ‘Liberibacter’ asiaticus that causes citrus greening disease. Because juvenile hormone (JH) plays an important role in adult and nymphal development, we studied the final steps in JH biosynthesis in D. citri . A putative JH acid methyltransferase ortholog gene (jmt D) and its cognate cDNA were identified by searching D. citri genome database. Expression analysis shows expression in all life stages. In adults, it is expressed in the head-thorax, (containing the corpora allata), and the abdomen (containing ovaries and male accessory glands). A 3D protein model identified the catalytic groove with catalytically active amino acids and the S-adenosyl methionine (SAM)-binding loop. The cDNA was expressed in Escherichia coli cells and the purified enzyme showed high preference for farnesoic acid (FA) and homoFA (k_cat of 0.752 × 10^–3 and 0.217 × 10^–3s^–1, respectively) as compared to JH acid I (JHA I) (cis/trans/cis; 2Z, 6E, 10cis), JHA III (2E, 6E,10cis), and JHA I (trans/cis/cis; 2E, 2Z, 10cis) (k_cat of 0.081 × 10^–3, 0.013 × 10^–3, and 0.003 × 10^–3s^–1, respectively). This suggests that this ortholog is a Dc FA-o-methyl transferase gene (fmtD), not a jmt D, and that JH biosynthesis in D. citri proceeds from FA to JH III through methyl farnesoate (MF). DcFA-o-MT does not require Ca²⁺, Mg²⁺ or Zn²⁺, however, Zn²⁺ (1 mM) completely inhibits the enzyme probably by binding H115 at the active groove. This represents the first purified FA-o-MT from Hemiptera with preferred biological activity for FA and not JHA.

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Research Organization:: Oak Ridge Associated Univ., Oak Ridge, TN (United States)

Sponsoring Organization:: USDOE; USDA; Agricultural Research Service (ARS)

Grant/Contract Number:: AC05-06OR23100

OSTI ID:: 1209195

Alternate ID(s):: OSTI ID: 1623670

Journal Information:: FEBS Open Bio, Vol. 5, Issue 1; ISSN 2211-5463

Publisher:: Federation of European Biochemical SocietiesCopyright Statement

Country of Publication:: United States

Language:: English

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Cited by: 9 works

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biochemistry & molecular biology
Diaphorina citri
juvenile hormone acid methyltransferase
farnesoic acid methyltransferase
3D modeling
gene expression