title: Genetic suppression analysis in novel vacuolar processing enzymes reveals their roles in controlling sugar accumulation in tomato fruits.
abstract: In plant cells, many vacuolar proteins are synthesized as precursors in the endoplasmic reticulum and are subsequently transported to the vacuole. These precursors are subject to post-translational modifications to allow the active mature forms to be produced. Vacuolar processing enzyme (VPE) has been identified as a family of cysteine proteases involved in protein maturation in the vacuole. In this study, novel VPE genes were isolated from tomato (Solanum lycopersicum), and they were designated SlVPE1-SlVPE5. Phylogenic analysis suggested that SlVPE1 and SlVPE2 were categorized as the seed coat type, SlVPE4 was categorized as the seed type, and both SlVPE3 and SlVPE5 were categorized as the vegetative type. Expression analysis demonstrated that these genes were expressed during fruit development, and that their expression profiles agreed with this classification. High VPE enzyme activity was observed during tomato fruit development; the enzyme activity was correlated with the SlVPE mRNA levels, indicating that the SlVPE encoded active VPE proteins. The total sugar content was higher in RNA interference (RNAi) lines compared with the control plants, suggesting negative roles for SlVPE in sugar accumulation. The quantitative expression analysis of each SlVPE gene in the RNAi lines suggested that the suppression of SlVPE5 probably had the strongest effect on the sugar accumulation observed. The suppression of SlVPE did not influence the total amino acid content, suggesting that the molecular targets of SlVPE were mainly involved in sugar accumulation.
authors.affiliation: Graduate School of Environmental Sciences, Gene Research Center, University of Tsukuba, 1-1-1 Tennodai, Tsukuba 305-8572. Japan.
authors.forename: Tohru
authors.initials: T
authors.lastname: Ariizumi
authors.forename: Kenji
authors.initials: K
authors.lastname: Higuchi
authors.forename: Shoko
authors.initials: S
authors.lastname: Arakaki
authors.forename: Tsunenori
authors.initials: T
authors.lastname: Sano
authors.forename: Erika
authors.initials: E
authors.lastname: Asamizu
authors.forename: Hiroshi
authors.initials: H
authors.lastname: Ezura
mesh_terms: D000596:Amino Acids / Q000378:metabolism; D050260:Carbohydrate Metabolism / Q000235:genetics*; D003546:Cysteine Endopeptidases / Q000235:genetics* / Q000378:metabolism; D005638:Fruit / Q000201:enzymology* / Q000235:genetics* / Q000254:growth & development; D015971:Gene Expression Regulation, Enzymologic; D018506:Gene Expression Regulation, Plant; D017930:Genes, Reporter; D005966:Glucuronidase / Q000378:metabolism; D007150:Immunohistochemistry; D018551:Solanum lycopersicum / Q000201:enzymology* / Q000235:genetics* / Q000254:growth & development; D010802:Phylogeny; D010940:Plant Proteins / Q000235:genetics / Q000378:metabolism; D030821:Plants, Genetically Modified; D034622:RNA Interference; D012333:RNA, Messenger / Q000235:genetics / Q000378:metabolism; D020133:Reverse Transcriptase Polymerase Chain Reaction; D013489:Suppression, Genetic; D013997:Time Factors
chemical_list: D000596:Amino Acids; D010940:Plant Proteins; D012333:RNA, Messenger; D005966:Glucuronidase; D003546:Cysteine Endopeptidases; C089278:vacuolar processing enzyme