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PMID: 9869425 Published · ppublish English Journal Article Research Support, Non-U.S. Gov't

Anthranilate N-hydroxycinnamoyl/benzoyltransferase gene from carnation: rapid elicitation of transcription and promoter analysis.

Plant molecular biology ·Vol. 38 ·No. 6 ·1998-12-00 ·Pages 1201-14

Yang Q, Grimmig B, Matern U

Abstract

Hydroxycinnamoyl/benzoyl-CoA:anthranilate N-hydroxycinnamoyl/benzoyltransferase (HCBT) catalyses the committed reaction of phytoalexin biosynthesis in carnation (Dianthus caryophyllus L.). Three HCBT cDNAs were cloned previously from suspension-cultured carnation cells that had been induced with fungal elicitor. A rapid, transient induction of hcbt transcripts upon elicitation, reaching maximal abundances within about 0.5 h and returning to basal levels within 4 h, suggested the involvement of unusual cis elements. A DNA fragment of 3.8 kb, spanning the hcbt2 gene with the 5'-flanking region of roughly 1.8 kb, was cloned from carnation plants. The gene encodes one long open reading frame lacking introns. The DNA sequence revealed a conserved TATA box, three elicitor response elements (EREs) and a 9 bp direct repeat as well as an interrupted direct repeat of 11 bp in the TATA distal region. EMSA revealed the binding of elicitor-inducible nuclear factors to the promoter region from -377 to -326 spanning two of the EREs, and their functional relevance was confirmed by transient expression assays of hcbt2 promoter-GUS reporter gene constructs in parsley protoplasts. Furthermore, an oligo(A) segment was present immediately preceding the start of translation (+140 to +150). Transient expression analysis demonstrated that the sequence upstream to -1157 at least is required in context with the 5'-UTR, particularly including the poly(A) segment, for strong expression and full elicitor induction of the hcbt2 gene. The results suggested that several sequence motifs scattered over a wide range of the 5'-flanking region and into the exonic sequence are responsible for the full elicitor regulation of the hcbt2 gene.

MeSH Terms
Acyltransferases/genetics,metabolism Base Sequence Cell Nucleus/metabolism Cells, Cultured Cloning, Molecular Conserved Sequence DNA, Complementary Gene Expression Regulation, Plant Molecular Sequence Data Plant Extracts/biosynthesis Plant Leaves Plants/enzymology,genetics Polymerase Chain Reaction Promoter Regions, Genetic Protein Biosynthesis Sequence Alignment Sequence Homology, Nucleic Acid Sesquiterpenes TATA Box Terpenes Transcription, Genetic
Chemicals
DNA, Complementary Plant Extracts Sesquiterpenes Terpenes phytoalexins Acyltransferases anthranilate N-hydroxycinnamoyl benzoyltransferase
Authors & Affiliations
3 authors, click to expand affiliations / ORCID
Yang Q
Institut für Biologie II, Lehrstuhl für Biochemie der Pflanzen, Universität Freiburg, Germany.
Grimmig B
Matern U
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Article Info
Journal
Plant molecular biology
Abbr.
Plant Mol Biol
ISSN
0167-4412
DOI
10.1023/a:1006003731919
Published
1998-12-00
Pages
1201-14
Language
English
Region
Netherlands
NLM ID
9106343
Subset
IM
Databases
GENBANK
Z98758
External Links
PubMed source DOI: 10.1023/a:1006003731919 View journal details
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