Sequence analysis of a "true" chalcone synthase (chs_H1) oligofamily from hop (Humulus lupulus L.) and PAP1 activation of chs_H1 in heterologous systems
Language English Country United States Media print
Document type Journal Article, Research Support, Non-U.S. Gov't
PubMed
17002429
DOI
10.1021/jf061785g
Knihovny.cz E-resources
- MeSH
- Acyltransferases chemistry genetics metabolism MeSH
- Enzyme Activation drug effects MeSH
- Anthocyanins biosynthesis MeSH
- Plants, Genetically Modified MeSH
- Humulus enzymology genetics MeSH
- Plant Leaves metabolism MeSH
- RNA, Messenger analysis MeSH
- Molecular Sequence Data MeSH
- Reverse Transcriptase Polymerase Chain Reaction MeSH
- Pancreatitis-Associated Proteins MeSH
- Arabidopsis Proteins MeSH
- Amino Acid Sequence MeSH
- Base Sequence MeSH
- Transcription Factors pharmacology MeSH
- Ultraviolet Rays MeSH
- Publication type
- Journal Article MeSH
- Research Support, Non-U.S. Gov't MeSH
- Names of Substances
- Acyltransferases MeSH
- Anthocyanins MeSH
- flavanone synthetase MeSH Browser
- RNA, Messenger MeSH
- PAP1 protein, Arabidopsis MeSH Browser
- Pancreatitis-Associated Proteins MeSH
- Arabidopsis Proteins MeSH
- REG3A protein, human MeSH Browser
- Transcription Factors MeSH
Screening of a cDNA library of the hop cv. Osvald's 72 and genomic cloning were used to isolate members of an oligofamily of chs_H1 genes that codetermine the biosynthesis of prenylated chalcones known to be valuable medicinal compounds present in hop (Humulus lupulus L.). chs_H1 oligofamily members showed more than 99% and 98% identity on nucleotide and amino acid levels, respectively, and retained all conserved amino acids that form the catalytic center characteristic for "true" chalcone synthases. The chs_H1 promoter exhibited low sequence variability in addition to conservation of all predicted cis-regulatory elements. Possible transactivation of the chs_H1 gene with the transcription factor PAP1 from Arabidopsis thaliana was assayed using Agrobacterium tumefaciens infiltrations of Nicotiana benthamiana and Petunia hybrida plants. Infiltration of N. benthamiana leaves with chs_H1 promoter/GUS chimeras led to a 24.8-fold increase of the GUS activity when coinfiltrated with the pap1 gene. Coinfiltration of the "native" chs_H1 gene with pap1 led to an increased accumulation of chs_H1 mRNA as observed by semiquantitative reverse transcription-polymerase chain reaction. Transgenic lines of P. hybrida expressing the pap1 gene showed unusual patterns of UV-A-inducible pigmentation and anthocyanin accumulation in parenchymatic and medulla cells. Infiltration of transgenic leaves of P. hybrida with chs_H1 and pap1 genes arranged as a tandem led to quick pigmentation within 12 h after UV-A irradiation. It is indicated that the chs_H1 promoter contains functional element(s) mediating an efficient response to PAP1 expression and UV-A irradiation. UV-A also induced chs_H1 mRNA and accumulation of flavonol glycosides in hop leaves. It can be expected that the PAP1 factor could significantly influence the expression of the chs_H1 oligofamily in transgenic hop and modify the hop metabolome.
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