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PMID: 9349252 Published · ppublish English Journal Article Research Support, Non-U.S. Gov't

Characterization of a gene encoding a DNA-binding protein that interacts in vitro with vascular specific cis elements of the phenylalanine ammonia-lyase promoter.

Plant molecular biology ·Vol. 35 ·No. 3 ·1997-10-00 ·Pages 281-91

Séguin A, Laible G, Leyva A, Dixon RA, Lamb CJ

Abstract

A study of the expression of a bean phenylalanine ammonia-lyase (PAL) promoter/beta-glucuronidase gene fusion in transgenic tobacco has shown that the PAL2 promoter has a modular organization. Expression of the PAL2 promoter in the vascular system involves positive and negative regulatory cis elements. Among these elements is an AC-rich motif implicated in xylem expression and a suppressing cis element for phloem expression. Using radiolabelled complementary oligonucleotides bearing the AC-rich motif, a cDNA clone encoding a DNA-binding protein has been isolated from a tobacco lambda gt11 expression library. This factor, named AC-rich binding factor (ACBF), showed binding specificity to the AC-rich region. The specificity of ACBF for the AC-rich region was also shown using a gel retardation assay with an ACBF recombinant protein extract. The deduced amino acid sequence from ACBF contains a long repeat of glutamine residues as found in well characterized transcription factors. Interestingly, ACBF shared sequence similarity to conserved amino acid motifs found in RNA-binding proteins. Genomic gel blot analysis indicated the presence of a small gene family of sequences related to ACBF within the tobacco nuclear genome. Analysis of tobacco mRNA using the ACBF cDNA as probe showed that while ACBF mRNA was present in all tissues examined, the highest transcript accumulation occurred in stem tissues. The functional characteristics of the AC-rich sequence were examined in transgenic tobacco. A heptamer of the AC-rich sequence, in front of a minimal 35S promoter from cauliflower mosaic virus (-46 to +4), conferred specific expression in xylem.

MeSH Terms
Amino Acid Sequence Base Sequence DNA, Complementary/isolation & purification DNA-Binding Proteins/genetics,metabolism,physiology Gene Expression Regulation, Plant Genes, Plant Genome, Plant Molecular Sequence Data Phenylalanine Ammonia-Lyase/genetics Plant Proteins/genetics,metabolism,physiology Plants, Genetically Modified Plants, Toxic Promoter Regions, Genetic Protein Binding/genetics RNA-Binding Proteins/genetics Recombinant Proteins/metabolism Regulatory Sequences, Nucleic Acid/physiology Tobacco
Chemicals
ACBF protein, tobacco DNA, Complementary DNA-Binding Proteins Plant Proteins RNA-Binding Proteins Recombinant Proteins Phenylalanine Ammonia-Lyase
Authors & Affiliations
5 authors, click to expand affiliations / ORCID
Séguin A
Plant Biology Laboratory, Salk Institute for Biological Studies, La Jolla, CA 92037, USA.
Laible G
Leyva A
Dixon R A
Lamb C J
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Article Info
Journal
Plant molecular biology
Abbr.
Plant Mol Biol
ISSN
0167-4412
DOI
10.1023/a:1005853404242
Published
1997-10-00
Pages
281-91
Language
English
Region
Netherlands
NLM ID
9106343
Subset
IM
Databases
GENBANK
U90212
External Links
PubMed source DOI: 10.1023/a:1005853404242 View journal details
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