Identification of a Bifunctional Maize C- and O-Glucosyltransferase

Ferreyra, Marı́a Lorena Falcone; Rodrı́guez, Eduardo; Casas, María Isabel; Labadié, Guillermo R.; Grotewold, Erich; Casati, Paula

doi:10.1074/jbc.m113.510040

Cited by 131 publications

(89 citation statements)

References 56 publications

Supporting

Mentioning

Contrasting

Unclassified

Order By: Relevance

“…This approach could be easily adapted to probe the activity of other metabolic enzymes. These studies, together with our previous characterization of CYP93G5 that converts flavanones (including naringenin and eriodictyol) to the respective 2-hydroxyflavanones (Morohashi et al, 2012) and UGT708A6 that C-glycosylates 2-hydroxyflavanones to the respective flavone-6-C-glucosides (Falcone Ferreyra et al, 2013), complete the maize maysin biosynthetic pathway.…”

Section: Discussionmentioning

confidence: 99%

“…Of the 336 gene models contained in the sm2 mapping interval ( Figure 3A; Supplemental Table 2), mRNA levels for nine genes were significantly higher in P1-rr compared with P1-ww silks (Supplemental Table 2). Sequence analyses from proteins encoded by these genes yielded UGT91L1 (GRMZM2G180283) as a putative UDP-glycosyltransferase (UGT) that clustered together with previously characterized UGTs that use flavonoid glycosides as substrate acceptors and catalyze the formation of sugar-Osugar links, including the rhamnosyl transferases from Petunia hybrida and Citrus maxima (Falcone Ferreyra et al, 2013). Moreover, in phylogenetic analyses of all putative maize UGTs, five additional UGTs clustered together with UGT91L1, but none of the genes encoding these UGTs are upregulated by P1 in silks, further supporting UGT91L1 as an Sm2 candidate (Supplemental Figure 3 and Supplemental Data Set 1).…”

Section: Identification and Characterization Of Sm2 Candidate Genesmentioning

confidence: 99%

“…Total protein extracts from yeast expressing At-RHM1 or empty pGZ25 vector and total protein extracts from Escherichia coli expressing UGT91L1 or empty pET28a vector were prepared. To obtain E. coli crude extracts, E. coli Rosetta 2 (DE3) cells expressing UGT91L1 were grown as previously described (Falcone Ferreyra et al, 2013). Cells were harvested by centrifugation, resuspended in 50 mM KPi, pH 7.5, with 1 mM PMSF, sonicated, and centrifuged at 12,000g for 15 min at 4°C.…”

Section: Rhamnosyl Transferase Activity Assaymentioning

confidence: 99%

“…The T7 pro :His 6 -UGT91L1 construct in pET28a was previously described (Falcone Ferreyra et al, 2013). The RHS1 (GRMZM2G031311) coding sequence was amplified from P1-rr silks using the OneStep RT-PCR kit (Qiagen).…”

Section: Cloning Of Ugt91l1 Rhs1 and At-rhm1mentioning

confidence: 99%

“…Subsequently, 2-hydroxyeriodictyol is C-glycosylated by the C-glucosyl transferase UGT708A6 (Falcone Ferreyra et al, 2013), and following a likely spontaneous dehydration reaction (Brazier-Hicks et al, 2009), isoorientin (luteolin 6-C-glucoside) is formed ( Figure 1). From the structures determined by McMullen et al (2004) using NMR, the conversion of isoorientin to maysin requires at least two additional enzymatic steps: the first one involving the incorporation of a rhamnose to the glucose moiety of isoorientin to form rhamnosylisoorientin (isoorientin 299-O-rhamnoside) and the second involving the dehydration of glucose to 4-keto-6-deoxy glucose to convert rhamnosylisoorientin into maysin (Figure 1).…”

Section: Introductionmentioning

confidence: 99%

See 4 more Smart Citations

Identification and Characterization of Maize salmon silks Genes Involved in Insecticidal Maysin Biosynthesis

et al. 2016

Self Cite

View full text Add to dashboard Cite

The century-old maize (Zea mays) salmon silks mutation has been linked to the absence of maysin. Maysin is a C-glycosyl flavone that, when present in silks, confers natural resistance to the maize earworm (Helicoverpa zea), which is one of the most damaging pests of maize in America. Previous genetic analyses predicted Pericarp Color1 (P1; R2R3-MYB transcription factor) to be epistatic to the sm mutation. Subsequent studies identified two loci as being capable of conferring salmon silks phenotypes, salmon silks1 (sm1) and sm2. Benefitting from available sm1 and sm2 mapping information and from knowledge of the genes regulated by P1, we describe here the molecular identification of the Sm1 and Sm2 gene products. Sm2 encodes a rhamnosyl transferase (UGT91L1) that uses isoorientin and UDP-rhamnose as substrates and converts them to rhamnosylisoorientin. Sm1 encodes a multidomain UDP-rhamnose synthase (RHS1) that converts UDP-glucose into UDP-Lrhamnose. Here, we demonstrate that RHS1 shows unexpected substrate plasticity in converting the glucose moiety in rhamnosylisoorientin to 4-keto-6-deoxy glucose, resulting in maysin. Both Sm1 and Sm2 are direct targets of P1, as demonstrated by chromatin immunoprecipitation experiments. The molecular characterization of Sm1 and Sm2 described here completes the maysin biosynthetic pathway, providing powerful tools for engineering tolerance to maize earworm in maize and other plants.

show abstract

Section: Discussionmentioning

confidence: 99%

Section: Identification and Characterization Of Sm2 Candidate Genesmentioning

confidence: 99%

Section: Rhamnosyl Transferase Activity Assaymentioning

confidence: 99%

Section: Cloning Of Ugt91l1 Rhs1 and At-rhm1mentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

See 3 more Smart Citations

Identification and Characterization of Maize salmon silks Genes Involved in Insecticidal Maysin Biosynthesis

et al. 2016

Self Cite

View full text Add to dashboard Cite

show abstract

Probing the Catalytic Promiscuity of a Regio‐ and Stereospecific C‐Glycosyltransferase from Mangifera indica

Chen

Wang

et al. 2015

Angewandte Chemie

View full text Add to dashboard Cite

The catalytic promiscuity of the novel benzophenone C-glycosyltransferase,M iCGT,w hich is involved in the biosynthesis of mangiferin from Mangifera indica, was explored. MiCGT exhibited ar obust capability to regio-and stereospecific C-glycosylation of 35 structurally diverse druglike scaffolds and simple phenolics with UDP-glucose,a nd also formed O-and N-glycosides.Moreover,MiCGT was able to generate C-xylosides with UDP-xylose.T he OGT-reversibility of MiCGT was also exploited to generate C-glucosides with simple sugar donor.T hree aryl-C-glycosides exhibited potent SGLT2 inhibitory activities with IC 50 values of 2.6 , 7.6 , and 7.6 10 À7 m,r espectively.T hese findings demonstrate for the first time the significant potential of an enzymatic approach to diversification through C-glycosidation of bioactive natural and unnatural products in drug discovery.

show abstract

Molecular and Structural Characterization of a Promiscuous C‐Glycosyltransferase from Trollius chinensis

Zhao

et al. 2019

Angewandte Chemie

View full text Add to dashboard Cite

Herein, the catalytic promiscuity of TcCGT1, anew C-glycosyltransferase (CGT) from the medicinal plant Trollius chinensis is explored. TcCGT1 could efficiently and regiospecifically catalyze the 8-C-glycosylation of 36 flavones and other flavonoids and could also catalyze the O-glycosylation of diverse phenolics.The crystal structure of TcCGT1 in complex with uridine diphosphate was determined at 1.85 resolution. Molecular docking revealed an ew model for the catalytic mechanism of TcCGT1, which is initiated by the spontaneous deprotonation of the substrate.T he spacious binding pocket explains the substrate promiscuity,and the binding pose of the substrate determines C-or O-glycosylation activity.S itedirected mutagenesis at two residues (I94E and G284K) switched C-to O-glycosylation. TcCGT1 is the first plant CGT with ac rystal structure and the first flavone 8-Cglycosyltransferase described. This provides abasis for designing efficient glycosylation biocatalysts.

show abstract

Identification of a Bifunctional Maize C- and O-Glucosyltransferase

Cited by 131 publications

References 56 publications

Identification and Characterization of Maize salmon silks Genes Involved in Insecticidal Maysin Biosynthesis

Identification and Characterization of Maize salmon silks Genes Involved in Insecticidal Maysin Biosynthesis

Probing the Catalytic Promiscuity of a Regio‐ and Stereospecific C‐Glycosyltransferase from Mangifera indica

Molecular and Structural Characterization of a Promiscuous C‐Glycosyltransferase from Trollius chinensis

Contact Info

Product

Resources

About