Cloning and expressing a highly functional and substrate specific farnesoic acid o‐methyltransferase from the Asian citrus psyllid (Diaphorina citri Kuwayama)
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 inD. citri. A...
| Main Authors: | , , , , , |
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| Format: | Article |
| Language: | English |
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Wiley
2015-01-01
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| Series: | FEBS Open Bio |
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| Online Access: | https://doi.org/10.1016/j.fob.2015.03.012 |
| _version_ | 1811185111654203392 |
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| author | Evelien Van Ekert Robert G. Shatters Pierre Rougé Charles A. Powell Guy Smagghe Dov Borovsky |
| author_facet | Evelien Van Ekert Robert G. Shatters Pierre Rougé Charles A. Powell Guy Smagghe Dov Borovsky |
| author_sort | Evelien Van Ekert |
| collection | DOAJ |
| description | 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 inD. citri. A putative JH acid methyltransferase ortholog gene (jmtD) and its cognate cDNA were identified by searchingD. 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 inEscherichia 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 aDcFA‐o‐methyl transferase gene (fmtD), not ajmtD, and that JH biosynthesis inD. 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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| institution | Directory Open Access Journal |
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| language | English |
| last_indexed | 2024-04-11T13:23:51Z |
| publishDate | 2015-01-01 |
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| spelling | doaj.art-bed2fd17cc1a4931902cfcf3fdee34c62022-12-22T04:22:07ZengWileyFEBS Open Bio2211-54632015-01-015126427510.1016/j.fob.2015.03.012Cloning and expressing a highly functional and substrate specific farnesoic acid o‐methyltransferase from the Asian citrus psyllid (Diaphorina citri Kuwayama)Evelien Van Ekert0Robert G. Shatters1Pierre Rougé2Charles A. Powell3Guy Smagghe4Dov Borovsky5USDA-ARS, Ft. Pierce, FL 34945, USAUSDA-ARS, Ft. Pierce, FL 34945, USAUniversité de Toulouse, UPS, Institut de Recherche pour le Développement (IRD), UMR 152 Pharma-Dev, Université Toulouse 3, Faculté des Sciences Pharmaceutiques, F-31062 Toulouse cedex 09, FranceIndian River Research and Education Center, University of Florida, FL 34945, USAUniversiteit Gent, Gent 9000, BelgiumUSDA-ARS, Ft. Pierce, FL 34945, USAThe 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 inD. citri. A putative JH acid methyltransferase ortholog gene (jmtD) and its cognate cDNA were identified by searchingD. 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 inEscherichia 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 aDcFA‐o‐methyl transferase gene (fmtD), not ajmtD, and that JH biosynthesis inD. 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.https://doi.org/10.1016/j.fob.2015.03.012Diaphorina citriJuvenile hormone acid methyltransferaseFarnesoic acid methyltransferase3D modelingGene expression |
| spellingShingle | Evelien Van Ekert Robert G. Shatters Pierre Rougé Charles A. Powell Guy Smagghe Dov Borovsky Cloning and expressing a highly functional and substrate specific farnesoic acid o‐methyltransferase from the Asian citrus psyllid (Diaphorina citri Kuwayama) FEBS Open Bio Diaphorina citri Juvenile hormone acid methyltransferase Farnesoic acid methyltransferase 3D modeling Gene expression |
| title | Cloning and expressing a highly functional and substrate specific farnesoic acid o‐methyltransferase from the Asian citrus psyllid (Diaphorina citri Kuwayama) |
| title_full | Cloning and expressing a highly functional and substrate specific farnesoic acid o‐methyltransferase from the Asian citrus psyllid (Diaphorina citri Kuwayama) |
| title_fullStr | Cloning and expressing a highly functional and substrate specific farnesoic acid o‐methyltransferase from the Asian citrus psyllid (Diaphorina citri Kuwayama) |
| title_full_unstemmed | Cloning and expressing a highly functional and substrate specific farnesoic acid o‐methyltransferase from the Asian citrus psyllid (Diaphorina citri Kuwayama) |
| title_short | Cloning and expressing a highly functional and substrate specific farnesoic acid o‐methyltransferase from the Asian citrus psyllid (Diaphorina citri Kuwayama) |
| title_sort | cloning and expressing a highly functional and substrate specific farnesoic acid o methyltransferase from the asian citrus psyllid diaphorina citri kuwayama |
| topic | Diaphorina citri Juvenile hormone acid methyltransferase Farnesoic acid methyltransferase 3D modeling Gene expression |
| url | https://doi.org/10.1016/j.fob.2015.03.012 |
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