Cloning and expression of solanidine UDP-glucose glucosyltransferase from potato.
A cDNA encoding solanidine glucosyltransferase (SGT) was isolated from potato. The cDNA was selected from a yeast expression library using a positive selection based on the higher toxicity of steroidal alkaloid aglycons relative to their associated glycosylated forms. The cDNA contained an open reading frame encoding a 56 kDa polypeptide with regions of similarity to previously characterized UDP-glucosyltransferases. The enzyme activity and reaction products of recombinant SGT in yeast were consistent with those observed for the endogenous enzyme from potato. SGT mRNA and protein accumulated in tubers in response to wounding. The time course for SGT mRNA accumulation paralleled that of 3-hydroxy-3-methylglutaryl-coenzymeA isoform 1 (hmg1) mRNA. Steady-state SGT mRNA levels also increased transiently upon wounding of leaves.
Moehs
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United States Department of Agriculture, Agricultural Research Service, Western Regional Research Center, Albany, CA 94710, USA.
Allen
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PV
Friedman
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Belknap
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WR
D000595:Amino Acid Sequence; D001483:Base Sequence; D002265:Carboxylic Ester Hydrolases; D003001:Cloning, Molecular / Q000379:methods; D004926:Escherichia coli / Q000235:genetics; D018506:Gene Expression Regulation, Plant / Q000502:physiology*; D005964:Glucosyltransferases / Q000235:genetics*; D006903:Hydroxymethylglutaryl CoA Reductases / Q000235:genetics; D025782:Hydroxymethylglutaryl-CoA-Reductases, NADP-dependent; D008969:Molecular Sequence Data; D010940:Plant Proteins / Q000235:genetics; D012333:RNA, Messenger / Q000032:analysis; D018749:RNA, Plant / Q000032:analysis; D011994:Recombinant Proteins; D012441:Saccharomyces cerevisiae / Q000187:drug effects / Q000235:genetics; D017422:Sequence Analysis, DNA; D012991:Solanaceous Alkaloids / Q000378:metabolism / Q000494:pharmacology; D011198:Solanum tuberosum / Q000201:enzymology / Q000235:genetics*; D013379:Substrate Specificity; D015533:Transcriptional Activation
D010940:Plant Proteins; D012333:RNA, Messenger; D018749:RNA, Plant; D011994:Recombinant Proteins; D012991:Solanaceous Alkaloids; C043222:patatin protein, Solanum tuberosum; D006903:Hydroxymethylglutaryl CoA Reductases; D025782:Hydroxymethylglutaryl-CoA-Reductases, NADP-dependent; D005964:Glucosyltransferases; C077995:solanidine glucosyltransferase; D002265:Carboxylic Ester Hydrolases