Regulation of plant genes specifically induced in nitrogen-fixing nodules: role of cis-acting elements and trans-acting factors in leghemoglobin gene expression

F. J. de Bruijn, G. Felix, B. Grunenberg, H. J. Hoffmann, B. Metz, P. Ratet, A. Simons-Schreier, L. Szabados, P. Welters, J. Schell

Research output: Contribution to journalArticle

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Abstract

Transgenic alfalfa plants harboring a gene fusion between the soybean leghemoglobin (lbc3) promoter region and the chloramphenicol acetyl transferase (cat) gene were used to determine the influence of rhizobial mutants on lb gene expression in nodules. The promoter region of the Sesbania rostrata g lb3 (Srglb3) leghemoglobin gene was examined for the presence of conserved motifs homologous to binding site 1 and 2 of the soybean lbc3 promoter region, found to interact with a trans-acting factor present in soybean nodule nuclear extracts (Jensen EO, Marcker KA, Schell J, de Bruijn FJ, EMBO J 7: 1265-1271, 1988). Subfragments of the S. rostrata g lb3 (Srg lb3) promoter region were examined for binding to trans-acting factors from nodule nuclear extracts. In addition to the binding sites previously identified (Metz BA, Welters P, Hoffmann HJ, Jensen EO, Schell J, de Bruijn FJ, Mol Gen Genet 214: 181-191), several other sites were found to interact with trans-acting factors. In most cases the same trans-acting factor(s) were shown to be involved. One fragment (202) was found to bind specifically to a different factor (protein) which was extremely heat-resistant (100°C). The appearance of this factor was shown to be developmentally regulated since the expected protein-DNA complexes were first observed around 12 days after infection, concomitant with the production of leghemoglobin proteins. Fragments of the Srg lb3 5′ upstream region were fused to the β-glucuronidase reporter gene with its own CAAT and TATA box region or those of the cauliflower mosaic virus 35S and nopaline synthase (nos) promoters. These constructs were used to generate transgenic Lotus corniculatus plants and their expression was measured in different plant tissues. The Srg lb3 CAAT and TATA box region was found to be required for nodule-specific expression and several upstream enhancer-type regions were identified.

Original languageEnglish
Pages (from-to)319-325
Number of pages7
JournalPlant Molecular Biology
Volume13
Issue number3
DOIs
Publication statusPublished - Sep 1989

Fingerprint

Leghemoglobin
leghemoglobin
Plant Genes
Trans-Activators
Sesbania rostrata
Genetic Promoter Regions
Gene expression
Nitrogen
Genes
Soybeans
promoter regions
Gene Expression
gene expression
TATA Box
nitrogen
TATA box
soybeans
genes
Sesbania
Binding Sites

Keywords

  • cis-acting elements
  • gene regulation
  • leghmoglobin
  • nodulins
  • trans-acting factors
  • transgenic plants

ASJC Scopus subject areas

  • Biochemistry
  • Biochemistry, Genetics and Molecular Biology(all)

Cite this

Regulation of plant genes specifically induced in nitrogen-fixing nodules : role of cis-acting elements and trans-acting factors in leghemoglobin gene expression. / de Bruijn, F. J.; Felix, G.; Grunenberg, B.; Hoffmann, H. J.; Metz, B.; Ratet, P.; Simons-Schreier, A.; Szabados, L.; Welters, P.; Schell, J.

In: Plant Molecular Biology, Vol. 13, No. 3, 09.1989, p. 319-325.

Research output: Contribution to journalArticle

de Bruijn, FJ, Felix, G, Grunenberg, B, Hoffmann, HJ, Metz, B, Ratet, P, Simons-Schreier, A, Szabados, L, Welters, P & Schell, J 1989, 'Regulation of plant genes specifically induced in nitrogen-fixing nodules: role of cis-acting elements and trans-acting factors in leghemoglobin gene expression', Plant Molecular Biology, vol. 13, no. 3, pp. 319-325. https://doi.org/10.1007/BF00025320
de Bruijn, F. J. ; Felix, G. ; Grunenberg, B. ; Hoffmann, H. J. ; Metz, B. ; Ratet, P. ; Simons-Schreier, A. ; Szabados, L. ; Welters, P. ; Schell, J. / Regulation of plant genes specifically induced in nitrogen-fixing nodules : role of cis-acting elements and trans-acting factors in leghemoglobin gene expression. In: Plant Molecular Biology. 1989 ; Vol. 13, No. 3. pp. 319-325.
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abstract = "Transgenic alfalfa plants harboring a gene fusion between the soybean leghemoglobin (lbc3) promoter region and the chloramphenicol acetyl transferase (cat) gene were used to determine the influence of rhizobial mutants on lb gene expression in nodules. The promoter region of the Sesbania rostrata g lb3 (Srglb3) leghemoglobin gene was examined for the presence of conserved motifs homologous to binding site 1 and 2 of the soybean lbc3 promoter region, found to interact with a trans-acting factor present in soybean nodule nuclear extracts (Jensen EO, Marcker KA, Schell J, de Bruijn FJ, EMBO J 7: 1265-1271, 1988). Subfragments of the S. rostrata g lb3 (Srg lb3) promoter region were examined for binding to trans-acting factors from nodule nuclear extracts. In addition to the binding sites previously identified (Metz BA, Welters P, Hoffmann HJ, Jensen EO, Schell J, de Bruijn FJ, Mol Gen Genet 214: 181-191), several other sites were found to interact with trans-acting factors. In most cases the same trans-acting factor(s) were shown to be involved. One fragment (202) was found to bind specifically to a different factor (protein) which was extremely heat-resistant (100°C). The appearance of this factor was shown to be developmentally regulated since the expected protein-DNA complexes were first observed around 12 days after infection, concomitant with the production of leghemoglobin proteins. Fragments of the Srg lb3 5′ upstream region were fused to the β-glucuronidase reporter gene with its own CAAT and TATA box region or those of the cauliflower mosaic virus 35S and nopaline synthase (nos) promoters. These constructs were used to generate transgenic Lotus corniculatus plants and their expression was measured in different plant tissues. The Srg lb3 CAAT and TATA box region was found to be required for nodule-specific expression and several upstream enhancer-type regions were identified.",
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