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

Journal Article · · FEBS Open Bio

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 (kcat 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) (kcat 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.

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, Journal Name: FEBS Open Bio Vol. 5 Journal Issue: 1; ISSN 2211-5463
Publisher:
Wiley Blackwell (John Wiley & Sons)Copyright Statement
Country of Publication:
Netherlands
Language:
English
Citation Metrics:
Cited by: 9 works
Citation information provided by
Web of Science

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Figures / Tables (11)