DNA binding mediated by the wheat HMGa protein: a novel instance of selectivity against alternating GC sequence.
Identifieur interne : 003445 ( Main/Exploration ); précédent : 003444; suivant : 003446DNA binding mediated by the wheat HMGa protein: a novel instance of selectivity against alternating GC sequence.
Auteurs : Y L Chua [Singapour] ; K H Pwee ; R M Kini ; C Y Leng ; P K HockSource :
- Plant molecular biology [ 0167-4412 ] ; 2001.
Descripteurs français
- KwdFr :
- ADN des plantes (métabolisme), Amorces ADN, Chromatographie en phase liquide à haute performance, Cinétique, Liaison aux protéines, Protéines HMG (métabolisme), Régions promotrices (génétique), Résonance plasmonique de surface, Séquence nucléotidique, Triticum (génétique), Triticum (métabolisme), Électrophorèse sur gel de polyacrylamide.
- MESH :
- génétique : Triticum.
- métabolisme : ADN des plantes, Protéines HMG, Triticum.
- Amorces ADN, Chromatographie en phase liquide à haute performance, Cinétique, Liaison aux protéines, Régions promotrices (génétique), Résonance plasmonique de surface, Séquence nucléotidique, Électrophorèse sur gel de polyacrylamide.
English descriptors
- KwdEn :
- MESH :
- chemical , metabolism : DNA, Plant, High Mobility Group Proteins.
- chemical : DNA Primers.
- genetics : Triticum.
- metabolism : Triticum.
- Base Sequence, Chromatography, High Pressure Liquid, Electrophoresis, Polyacrylamide Gel, Kinetics, Promoter Regions, Genetic, Protein Binding, Surface Plasmon Resonance.
Abstract
The high-mobility-group (HMG) chromosomal protein wheat HMGa was purified to homogeneity and tested for its binding characteristics to double-stranded DNA. Wheat HMGa was able to bind to P268, an A/T-rich fragment derived from the pea plastocyanin gene promoter, producing a small mobility shift in gel retardation assays where the bound complex was sensitive to addition of proteinase K but resistant to heat treatment of the protein, consistent with the identity of wheat HMGa as a putative HMG-I/Y protein. Gel retardation assays and southwestern hybridization analysis revealed that wheat HMGa could selectively interact with the DNA polynucleotides poly(dA).poly(dT), poly(dAdT).poly(dAdT), and poly(dG).poly(dC), but not with poly(dGdC).poly(dGdC). Surface plasmon resonance analysis determined the kinetic and affinity constants of sensor chip-immobilized wheat HMGa for double-stranded DNA 10-mers, revealing a good affinity of the protein for various dinucleotide combinations, except that of alternating GC sequence. Thus contrary to prior reports of a selectivity of wheat HMGa for A/T-rich DNA, the protein appears to be able to interact with sequences containing guanine and cytosine residues as well, except where G/C residues alternate directly in the primary sequence.
DOI: 10.1023/a:1010696604330
PubMed: 11442059
Affiliations:
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Le document en format XML
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type="wicri:Area/Main/Curation">003445</idno><idno type="wicri:Area/Main/Exploration">003445</idno></publicationStmt><sourceDesc><biblStruct><analytic><title xml:lang="en">DNA binding mediated by the wheat HMGa protein: a novel instance of selectivity against alternating GC sequence.</title><author><name sortKey="Chua, Y L" sort="Chua, Y L" uniqKey="Chua Y" first="Y L" last="Chua">Y L Chua</name><affiliation wicri:level="4"><nlm:affiliation>Plant Molecular Biology Laboratory, National University of Singapore, Republic of Singapore.</nlm:affiliation><orgName type="university">Université nationale de Singapour</orgName><country>Singapour</country></affiliation></author><author><name sortKey="Pwee, K H" sort="Pwee, K H" uniqKey="Pwee K" first="K H" last="Pwee">K H Pwee</name></author><author><name sortKey="Kini, R M" sort="Kini, R M" uniqKey="Kini R" first="R M" last="Kini">R M Kini</name></author><author><name sortKey="Leng, C Y" sort="Leng, C Y" uniqKey="Leng C" first="C Y" last="Leng">C Y Leng</name></author><author><name sortKey="Hock, P K" sort="Hock, P K" uniqKey="Hock P" first="P K" last="Hock">P K Hock</name></author></analytic><series><title level="j">Plant molecular biology</title><idno type="ISSN">0167-4412</idno><imprint><date when="2001" type="published">2001</date></imprint></series></biblStruct></sourceDesc></fileDesc><profileDesc><textClass><keywords scheme="KwdEn" xml:lang="en"><term>Base Sequence</term><term>Chromatography, High Pressure Liquid</term><term>DNA Primers</term><term>DNA, Plant (metabolism)</term><term>Electrophoresis, Polyacrylamide Gel</term><term>High Mobility Group Proteins (metabolism)</term><term>Kinetics</term><term>Promoter Regions, Genetic</term><term>Protein Binding</term><term>Surface Plasmon Resonance</term><term>Triticum (genetics)</term><term>Triticum (metabolism)</term></keywords><keywords scheme="KwdFr" xml:lang="fr"><term>ADN des plantes (métabolisme)</term><term>Amorces ADN</term><term>Chromatographie en phase liquide à haute performance</term><term>Cinétique</term><term>Liaison aux protéines</term><term>Protéines HMG (métabolisme)</term><term>Régions promotrices (génétique)</term><term>Résonance plasmonique de surface</term><term>Séquence nucléotidique</term><term>Triticum (génétique)</term><term>Triticum (métabolisme)</term><term>Électrophorèse sur gel de polyacrylamide</term></keywords><keywords scheme="MESH" type="chemical" qualifier="metabolism" xml:lang="en"><term>DNA, Plant</term><term>High Mobility Group Proteins</term></keywords><keywords scheme="MESH" type="chemical" xml:lang="en"><term>DNA Primers</term></keywords><keywords scheme="MESH" qualifier="genetics" xml:lang="en"><term>Triticum</term></keywords><keywords scheme="MESH" qualifier="génétique" xml:lang="fr"><term>Triticum</term></keywords><keywords scheme="MESH" qualifier="metabolism" xml:lang="en"><term>Triticum</term></keywords><keywords scheme="MESH" qualifier="métabolisme" xml:lang="fr"><term>ADN des plantes</term><term>Protéines HMG</term><term>Triticum</term></keywords><keywords scheme="MESH" xml:lang="en"><term>Base Sequence</term><term>Chromatography, High Pressure Liquid</term><term>Electrophoresis, Polyacrylamide Gel</term><term>Kinetics</term><term>Promoter Regions, Genetic</term><term>Protein Binding</term><term>Surface Plasmon Resonance</term></keywords><keywords scheme="MESH" xml:lang="fr"><term>Amorces ADN</term><term>Chromatographie en phase liquide à haute performance</term><term>Cinétique</term><term>Liaison aux protéines</term><term>Régions promotrices (génétique)</term><term>Résonance plasmonique de surface</term><term>Séquence nucléotidique</term><term>Électrophorèse sur gel de polyacrylamide</term></keywords></textClass></profileDesc></teiHeader><front><div type="abstract" xml:lang="en">The high-mobility-group (HMG) chromosomal protein wheat HMGa was purified to homogeneity and tested for its binding characteristics to double-stranded DNA. Wheat HMGa was able to bind to P268, an A/T-rich fragment derived from the pea plastocyanin gene promoter, producing a small mobility shift in gel retardation assays where the bound complex was sensitive to addition of proteinase K but resistant to heat treatment of the protein, consistent with the identity of wheat HMGa as a putative HMG-I/Y protein. Gel retardation assays and southwestern hybridization analysis revealed that wheat HMGa could selectively interact with the DNA polynucleotides poly(dA).poly(dT), poly(dAdT).poly(dAdT), and poly(dG).poly(dC), but not with poly(dGdC).poly(dGdC). Surface plasmon resonance analysis determined the kinetic and affinity constants of sensor chip-immobilized wheat HMGa for double-stranded DNA 10-mers, revealing a good affinity of the protein for various dinucleotide combinations, except that of alternating GC sequence. Thus contrary to prior reports of a selectivity of wheat HMGa for A/T-rich DNA, the protein appears to be able to interact with sequences containing guanine and cytosine residues as well, except where G/C residues alternate directly in the primary sequence.</div></front></TEI><affiliations><list><country><li>Singapour</li></country><orgName><li>Université nationale de Singapour</li></orgName></list><tree><noCountry><name sortKey="Hock, P K" sort="Hock, P K" uniqKey="Hock P" first="P K" last="Hock">P K Hock</name><name sortKey="Kini, R M" sort="Kini, R M" uniqKey="Kini R" first="R M" last="Kini">R M Kini</name><name sortKey="Leng, C Y" sort="Leng, C Y" uniqKey="Leng C" first="C Y" last="Leng">C Y Leng</name><name sortKey="Pwee, K H" sort="Pwee, K H" uniqKey="Pwee K" first="K H" last="Pwee">K H Pwee</name></noCountry><country name="Singapour"><noRegion><name sortKey="Chua, Y L" sort="Chua, Y L" uniqKey="Chua Y" first="Y L" last="Chua">Y L Chua</name></noRegion></country></tree></affiliations></record>Pour manipuler ce document sous Unix (Dilib)
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