Proteins that bind telomeric DNA modulate the structure of chromosome ends and control telomere function and maintenance. It has been shown that AtTRB (Arabidopsis thaliana telomere-repeat-binding factor) proteins from the SMH (single-Myb-histone) family selectively bind double-stranded telomeric DNA and interact with the telomeric protein AtPOT1b (A. thaliana protection of telomeres 1b), which is involved in telomere capping. In the present study, we performed the first quantitative DNA-binding study of this plant-specific family of proteins. Interactions of full-length proteins AtTRB1 and AtTRB3 with telomeric DNA were analysed by electrophoretic mobility-shift assay, fluorescence anisotropy and surface plasmon resonance to reveal their binding stoichiometry and kinetics. Kinetic analyses at different salt conditions enabled us to estimate the electrostatic component of binding and explain different affinities of the two proteins to telomeric DNA. On the basis of available data, a putative model explaining the binding stoichiometry and the protein arrangement on telomeric DNA is presented.
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Research Article|
March 13 2009
Single-Myb-histone proteins from Arabidopsis thaliana: a quantitative study of telomere-binding specificity and kinetics
Ctirad Hofr;
Ctirad Hofr
1
*Department of Functional Genomics and Proteomics, Institute of Experimental Biology, Faculty of Science, Masaryk University, CZ-62500 Brno, Czech Republic
1Correspondence may be addressed to either of these authors (email hofr@sci.muni.cz or fajkus@sci.muni.cz).
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Pavla Šultesová;
Pavla Šultesová
*Department of Functional Genomics and Proteomics, Institute of Experimental Biology, Faculty of Science, Masaryk University, CZ-62500 Brno, Czech Republic
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Michal Zimmermann;
Michal Zimmermann
*Department of Functional Genomics and Proteomics, Institute of Experimental Biology, Faculty of Science, Masaryk University, CZ-62500 Brno, Czech Republic
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Iva Mozgová;
Iva Mozgová
*Department of Functional Genomics and Proteomics, Institute of Experimental Biology, Faculty of Science, Masaryk University, CZ-62500 Brno, Czech Republic
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Petra Procházková Schrumpfová;
Petra Procházková Schrumpfová
*Department of Functional Genomics and Proteomics, Institute of Experimental Biology, Faculty of Science, Masaryk University, CZ-62500 Brno, Czech Republic
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Michaela Wimmerová;
Michaela Wimmerová
†National Centre for Biomolecular Research and Department of Biochemistry, Faculty of Science, Masaryk University, CZ-61137 Brno, Czech Republic
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Jiří Fajkus
Jiří Fajkus
1
*Department of Functional Genomics and Proteomics, Institute of Experimental Biology, Faculty of Science, Masaryk University, CZ-62500 Brno, Czech Republic
‡Laboratory of DNA–Molecular Complexes, Institute of Biophysics, Czech Academy of Sciences, CZ-61265 Brno, Czech Republic
1Correspondence may be addressed to either of these authors (email hofr@sci.muni.cz or fajkus@sci.muni.cz).
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Publisher: Portland Press Ltd
Received:
November 07 2008
Revision Received:
December 18 2008
Accepted:
December 22 2008
Accepted Manuscript online:
December 22 2008
Online ISSN: 1470-8728
Print ISSN: 0264-6021
© The Authors Journal compilation © 2009 Biochemical Society
2009
Biochem J (2009) 419 (1): 221–230.
Article history
Received:
November 07 2008
Revision Received:
December 18 2008
Accepted:
December 22 2008
Accepted Manuscript online:
December 22 2008
Citation
Ctirad Hofr, Pavla Šultesová, Michal Zimmermann, Iva Mozgová, Petra Procházková Schrumpfová, Michaela Wimmerová, Jiří Fajkus; Single-Myb-histone proteins from Arabidopsis thaliana: a quantitative study of telomere-binding specificity and kinetics. Biochem J 1 April 2009; 419 (1): 221–230. doi: https://doi.org/10.1042/BJ20082195
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