Details
| Original language | English |
|---|---|
| Pages (from-to) | 2149-57 |
| Number of pages | 9 |
| Journal | Biochemistry |
| Volume | 42 |
| Issue number | 7 |
| Publication status | Published - 25 Feb 2003 |
Abstract
The expression of genes involved in C(4) photosynthesis in maize is under tight tissue-specific and light-dependent control. There is strong evidence that this control is at least in part brought about by DOF transcription factors binding to the respective promoters. We analyzed the interaction of DOF1 and DOF2 proteins with a functional and a cryptic endogenous binding site derived from the maize phosphoenolpyruvate carboxylase promoter (-300 bp region) in the nucleosomal context. Various DNA fragments comprising this promoter region were reconstituted into mononucleosomes from purified components, resulting in different positions of the DOF binding sites on the nucleosome surface. Binding of recombinant transcription factors to the different types of nucleosomes was examined using electrophoretic mobility shift assays. Changing the translational position of the binding site on the nucleosome surface strongly affected the efficiency of the interaction with the DOF factors. Deletion of individual recognition motifs revealed a positive impact of DOF protein binding to the main binding site on interactions with the cryptic binding site. The addition of the chromosomal high-mobility group (HMG) protein HMGB5 to the binding reaction mixture facilitated nucleosome binding of the transcription factor independent from the position of the recognition sites. The relevance of the data for the activation of the promoter in vivo is discussed.
Keywords
- Binding Sites, DNA, Plant/chemistry, DNA-Binding Proteins/chemistry, Electrophoretic Mobility Shift Assay/methods, HMGB Proteins/chemistry, Nucleosomes/chemistry, Plant Proteins/chemistry, Protein Binding, Structure-Activity Relationship, Transcription Factors/chemistry, Zea mays/chemistry
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In: Biochemistry, Vol. 42, No. 7, 25.02.2003, p. 2149-57.
Research output: Contribution to journal › Article › Research › peer review
}
TY - JOUR
T1 - The interaction of DOF transcription factors with nucleosomes depends on the positioning of the binding site and is facilitated by maize HMGB5
AU - Cavalar, Markus
AU - Möller, Claudia
AU - Offermann, Sascha
AU - Krohn, Nicholas M
AU - Grasser, Klaus D
AU - Peterhänsel, Christoph
PY - 2003/2/25
Y1 - 2003/2/25
N2 - The expression of genes involved in C(4) photosynthesis in maize is under tight tissue-specific and light-dependent control. There is strong evidence that this control is at least in part brought about by DOF transcription factors binding to the respective promoters. We analyzed the interaction of DOF1 and DOF2 proteins with a functional and a cryptic endogenous binding site derived from the maize phosphoenolpyruvate carboxylase promoter (-300 bp region) in the nucleosomal context. Various DNA fragments comprising this promoter region were reconstituted into mononucleosomes from purified components, resulting in different positions of the DOF binding sites on the nucleosome surface. Binding of recombinant transcription factors to the different types of nucleosomes was examined using electrophoretic mobility shift assays. Changing the translational position of the binding site on the nucleosome surface strongly affected the efficiency of the interaction with the DOF factors. Deletion of individual recognition motifs revealed a positive impact of DOF protein binding to the main binding site on interactions with the cryptic binding site. The addition of the chromosomal high-mobility group (HMG) protein HMGB5 to the binding reaction mixture facilitated nucleosome binding of the transcription factor independent from the position of the recognition sites. The relevance of the data for the activation of the promoter in vivo is discussed.
AB - The expression of genes involved in C(4) photosynthesis in maize is under tight tissue-specific and light-dependent control. There is strong evidence that this control is at least in part brought about by DOF transcription factors binding to the respective promoters. We analyzed the interaction of DOF1 and DOF2 proteins with a functional and a cryptic endogenous binding site derived from the maize phosphoenolpyruvate carboxylase promoter (-300 bp region) in the nucleosomal context. Various DNA fragments comprising this promoter region were reconstituted into mononucleosomes from purified components, resulting in different positions of the DOF binding sites on the nucleosome surface. Binding of recombinant transcription factors to the different types of nucleosomes was examined using electrophoretic mobility shift assays. Changing the translational position of the binding site on the nucleosome surface strongly affected the efficiency of the interaction with the DOF factors. Deletion of individual recognition motifs revealed a positive impact of DOF protein binding to the main binding site on interactions with the cryptic binding site. The addition of the chromosomal high-mobility group (HMG) protein HMGB5 to the binding reaction mixture facilitated nucleosome binding of the transcription factor independent from the position of the recognition sites. The relevance of the data for the activation of the promoter in vivo is discussed.
KW - Binding Sites
KW - DNA, Plant/chemistry
KW - DNA-Binding Proteins/chemistry
KW - Electrophoretic Mobility Shift Assay/methods
KW - HMGB Proteins/chemistry
KW - Nucleosomes/chemistry
KW - Plant Proteins/chemistry
KW - Protein Binding
KW - Structure-Activity Relationship
KW - Transcription Factors/chemistry
KW - Zea mays/chemistry
U2 - 10.1021/bi026761r
DO - 10.1021/bi026761r
M3 - Article
C2 - 12590604
VL - 42
SP - 2149
EP - 2157
JO - Biochemistry
JF - Biochemistry
SN - 0006-2960
IS - 7
ER -