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Characterisation of cis elements required for vascular expression of the cinnamoyl CoA reductase gene and for protein-DNA complex formation

Eric Lacombe, Jan Van Doorsselaere UGent, Wout Boerjan UGent, Alain M Boudet and Jacqueline Grima-Pettenati (2000) PLANT JOURNAL. 23(5). p.663-676
abstract
Cinnamoyl-CoA reductase (CCR) catalyses the first specific step in the biosynthesis of monolignols, the monomeric units of lignins. We examined the developmental regulation of the Eucalyptus gunnii EgCCR promoter by analysing the expression of EgCCR-GUS fusions in tobacco. EgCCR promoter activity was strongest in lignified organs (stems and roots) consistent with the EgCCR mRNA level in these organs. Histochemical analysis showed expression in vascular tissues (cambium, young differentiating xylem, ray cells, internal and external phloem) of stems and roots in agreement with in situ hybridization data. Promoter deletion analysis and gain-of-function experiments identified the sequences between positions -119 and -77 as necessary and sufficient for expression in vascular tissues of stems. Electrophoretic mobility-shift assays showed that this region is specifically recognized by nuclear proteins present in tobacco stems, giving rise to two retarded complexes, LMC1 and LMC2. Using overlapping EgCCR fragments and mutated oligonucleotides as competitors in gel-shift assays, two DNA-protein interaction sites were mapped. Finally, the role of protein-protein interactions in the formation of the LMC1 and LMC2 complexes was investigated using the detergent sodium deoxycholate, and protein fractionation onto a heparin Sepharose column.
Please use this url to cite or link to this publication:
author
organization
year
type
journalArticle (original)
publication status
published
subject
keyword
lignin, cinnamoyl CoA reductase, xylem, MYB factors, Eucalyptus, tobacco, AMMONIA-LYASE GENE, ELONGATION-FACTOR EF-1-ALPHA, TISSUE-SPECIFIC EXPRESSION, CELL-SPECIFIC ACTIVITY, PLANT DEFENSE GENES, TRANSGENIC TOBACCO, PHENYLPROPANOID METABOLISM, ARABIDOPSIS-THALIANA, ACTING ELEMENTS, ALCOHOL-DEHYDROGENASE
journal title
PLANT JOURNAL
Plant J.
volume
23
issue
5
pages
663 - 676
Web of Science type
Article
Web of Science id
000089401300010
ISSN
0960-7412
DOI
10.1046/j.1365-313x.2000.00838.x
language
English
UGent publication?
yes
classification
A1
copyright statement
I have transferred the copyright for this publication to the publisher
id
172538
handle
http://hdl.handle.net/1854/LU-172538
date created
2004-01-14 13:40:00
date last changed
2016-12-19 15:38:43
@article{172538,
  abstract     = {Cinnamoyl-CoA reductase (CCR) catalyses the first specific step in the biosynthesis of monolignols, the monomeric units of lignins. We examined the developmental regulation of the Eucalyptus gunnii EgCCR promoter by analysing the expression of EgCCR-GUS fusions in tobacco. EgCCR promoter activity was strongest in lignified organs (stems and roots) consistent with the EgCCR mRNA level in these organs. Histochemical analysis showed expression in vascular tissues (cambium, young differentiating xylem, ray cells, internal and external phloem) of stems and roots in agreement with in situ hybridization data. Promoter deletion analysis and gain-of-function experiments identified the sequences between positions -119 and -77 as necessary and sufficient for expression in vascular tissues of stems. Electrophoretic mobility-shift assays showed that this region is specifically recognized by nuclear proteins present in tobacco stems, giving rise to two retarded complexes, LMC1 and LMC2. Using overlapping EgCCR fragments and mutated oligonucleotides as competitors in gel-shift assays, two DNA-protein interaction sites were mapped. Finally, the role of protein-protein interactions in the formation of the LMC1 and LMC2 complexes was investigated using the detergent sodium deoxycholate, and protein fractionation onto a heparin Sepharose column.},
  author       = {Lacombe, Eric and Van Doorsselaere, Jan and Boerjan, Wout and Boudet, Alain M and Grima-Pettenati, Jacqueline},
  issn         = {0960-7412},
  journal      = {PLANT JOURNAL},
  keyword      = {lignin,cinnamoyl CoA reductase,xylem,MYB factors,Eucalyptus,tobacco,AMMONIA-LYASE GENE,ELONGATION-FACTOR EF-1-ALPHA,TISSUE-SPECIFIC EXPRESSION,CELL-SPECIFIC ACTIVITY,PLANT DEFENSE GENES,TRANSGENIC TOBACCO,PHENYLPROPANOID METABOLISM,ARABIDOPSIS-THALIANA,ACTING ELEMENTS,ALCOHOL-DEHYDROGENASE},
  language     = {eng},
  number       = {5},
  pages        = {663--676},
  title        = {Characterisation of cis elements required for vascular expression of the cinnamoyl CoA reductase gene and for protein-DNA complex formation},
  url          = {http://dx.doi.org/10.1046/j.1365-313x.2000.00838.x},
  volume       = {23},
  year         = {2000},
}

Chicago
Lacombe, Eric, Jan Van Doorsselaere, Wout Boerjan, Alain M Boudet, and Jacqueline Grima-Pettenati. 2000. “Characterisation of Cis Elements Required for Vascular Expression of the Cinnamoyl CoA Reductase Gene and for protein-DNA Complex Formation.” Plant Journal 23 (5): 663–676.
APA
Lacombe, E., Van Doorsselaere, J., Boerjan, W., Boudet, A. M., & Grima-Pettenati, J. (2000). Characterisation of cis elements required for vascular expression of the cinnamoyl CoA reductase gene and for protein-DNA complex formation. PLANT JOURNAL, 23(5), 663–676.
Vancouver
1.
Lacombe E, Van Doorsselaere J, Boerjan W, Boudet AM, Grima-Pettenati J. Characterisation of cis elements required for vascular expression of the cinnamoyl CoA reductase gene and for protein-DNA complex formation. PLANT JOURNAL. 2000;23(5):663–76.
MLA
Lacombe, Eric, Jan Van Doorsselaere, Wout Boerjan, et al. “Characterisation of Cis Elements Required for Vascular Expression of the Cinnamoyl CoA Reductase Gene and for protein-DNA Complex Formation.” PLANT JOURNAL 23.5 (2000): 663–676. Print.