title: Light response and potential interacting proteins of a grape flavonoid 3'-hydroxylase gene promoter.
abstract: Flavonoid 3'-hydroxylase (F3'H), a member of cytochrome P450 protein family, introduces B-ring hydroxyl group in the 3' position of the flavonoid. In this study, the cDNA sequence of a F3'H gene (VviF3'H), which contains an open reading frame of 1530 bp encoding a polypeptide of 509 amino acids, was cloned and characterized from Vitis vinifera L. cv. Cabernet Sauvignon. VviF3'H showed high homology to known F3'H genes, especially F3'Hs from the V. vinifera reference genome (Pinot Noir) and lotus. Expression profiling analysis using real-time PCR revealed that VviF3'H was ubiquitously expressed in all tested tissues including berries, leaves, flowers, roots, stems and tendrils, suggesting its important physiological role in plant growth and development. Moreover, the transcript level of VviF3'H gene in grape berries was relatively higher at early developmental stages and gradually decreased during véraison, and then increased in the mature phase. In addition, the promoter of VviF3'H was isolated by using TAIL-PCR. Yeast one-hybrid screening of the Cabernet Sauvignon cDNA library and subsequent in vivo/vitro validations revealed the interaction between VviF3'H promoter and several transcription factors, including members of HD-Zip, NAC, MYB and EIN families. A transcriptional regulation mechanism of VviF3'H expression is proposed for the first time.
authors.affiliation: Center for Viticulture and Enology, College of Food Science and Nutritional Engineering, China Agricultural University, Beijing, 100083, China.
authors.forename: Run-Ze
authors.initials: RZ
authors.lastname: Sun
authors.affiliation: Center for Viticulture and Enology, College of Food Science and Nutritional Engineering, China Agricultural University, Beijing, 100083, China.
authors.forename: Qiu-Hong
authors.initials: QH
authors.lastname: Pan
authors.affiliation: Center for Viticulture and Enology, College of Food Science and Nutritional Engineering, China Agricultural University, Beijing, 100083, China.
authors.forename: Chang-Qing
authors.initials: CQ
authors.lastname: Duan
authors.affiliation: Center for Viticulture and Enology, College of Food Science and Nutritional Engineering, China Agricultural University, Beijing, 100083, China. Electronic address: jun_wang@cau.edu.cn.
authors.forename: Jun
authors.initials: J
authors.lastname: Wang
mesh_terms: D000595:Amino Acid Sequence; D001483:Base Sequence; D003577:Cytochrome P-450 Enzyme System / Q000235:genetics / Q000378:metabolism*; D005419:Flavonoids / Q000378:metabolism*; D035264:Flowers / Q000201:enzymology / Q000235:genetics / Q000528:radiation effects; D005638:Fruit / Q000201:enzymology / Q000235:genetics / Q000528:radiation effects; D015971:Gene Expression Regulation, Enzymologic; D018506:Gene Expression Regulation, Plant; D008969:Molecular Sequence Data; D010802:Phylogeny; D018515:Plant Leaves / Q000201:enzymology / Q000235:genetics / Q000528:radiation effects; D010940:Plant Proteins / Q000235:genetics / Q000378:metabolism*; D018517:Plant Roots / Q000201:enzymology / Q000235:genetics / Q000528:radiation effects; D018547:Plant Stems / Q000201:enzymology / Q000235:genetics / Q000528:radiation effects; D011401:Promoter Regions, Genetic / Q000235:genetics; D016415:Sequence Alignment; D017422:Sequence Analysis, DNA; D014157:Transcription Factors / Q000235:genetics / Q000378:metabolism; D027843:Vitis / Q000201:enzymology* / Q000235:genetics / Q000528:radiation effects
chemical_list: D005419:Flavonoids; D010940:Plant Proteins; D014157:Transcription Factors; D003577:Cytochrome P-450 Enzyme System; C038888:flavonoid 3'-hydroxylase
keywords: Light sensitive; Promoter elements; Transcription factor; Vitis vinifera; Yeast one-hybrid