Application of fluorescence anisotropy to monitor protein binding to telomeric DNA

ZIMMERMANN, Michal, Ctirad HOFR, Pavla ŠULTESOVÁ, Iva MOZGOVÁ, Petra PROCHÁZKOVÁ SCHRUMPFOVÁ and Jiří FAJKUS. Application of fluorescence anisotropy to monitor protein binding to telomeric DNA. In FluoroFest Prague 2009. 2009.

Basic information

• Original name: Application of fluorescence anisotropy to monitor protein binding to telomeric DNA
• Name in Czech: Sledování vazby proteinů a telomerické DNA pomocí měření anizotropie fluorescence
• Authors: ZIMMERMANN, Michal (203 Czech Republic), Ctirad HOFR (203 Czech Republic), Pavla ŠULTESOVÁ (203 Czech Republic), Iva MOZGOVÁ (203 Czech Republic), Petra PROCHÁZKOVÁ SCHRUMPFOVÁ (203 Czech Republic) and Jiří FAJKUS (203 Czech Republic, guarantor).
• Edition: FluoroFest Prague 2009, 2009.

Other information

• Original language: English
• Type of outcome: Conference abstract
• Field of Study: Genetics and molecular biology
• Country of publisher: United States of America
• Confidentiality degree: is not subject to a state or trade secret
• RIV identification code: RIV/00216224:14310/09:00029388
• Organization unit: Faculty of Science
• Keywords in English: Protein-DNA interaction; Arabidopsis thaliana; SMH protein; fluorescence anisotropy; electrophoretic mobility shift assay
• Tags: Arabidopsis thaliana, electrophoretic mobility shift assay, fluorescence anisotropy, Protein-DNA interaction, SMH protein

Abstract

Using fluorescence anisotropy, we were able to compare the affinity of AtTRB proteins to fluorescently labeled DNA fragments harboring specific and nonspecific sequences and also to determine the optimal binding site by varying the size of the DNA fragments. Moreover, because fluorescence anisotropy enables the measurements to be performed in variety of experimental conditions, we managed to measure how the affinity depends on ionic strength. This allowed us to determine the electrostatic and nonelectrostatic contributions to the overall binding affinity of proteins to telomeric DNA. Based on these findings, a putative functional model of the complex between AtTRB proteins and telomeric DNA was constructed.

Abstract (in Czech)

Technikou měření anizotropie fluorescence byla určena afinita AtTRB proteinů k fluorescenčně značené DNA nesoucí specifické a nespecifické sekvence a zároveň byla zjištěna délka optimálního vazebného místa pro tyto proteiny. Protože anizotropie fluorescence umožňuje provádět experimenty v celé řadě podmínek, byla změřena závislost afinity na iontové síle. To umožnilo stanovit podíl elektrostatických a neelektrostatických sil na celkové afinitě. Na základě získaných výsledků byl nově navržen možný funkční model vytváření komplexu proteinů AtTRB a telomerové DNA.

Links

• GD204/08/H054, research and development project: Name: Molekulární mechanismy proliferace a diferenciace buněk

Investor: Czech Science Foundation, Molecular mechanisms of the cell proliferation and differentiation

• GP521/08/P452, research and development project: Name: Charakterizace a funkční diverzita proteinů rodiny SMH, která je specifická pro rostliny

Investor: Czech Science Foundation, Characterisation of functional diversity of the plant-specific SMH family of proteins

• MSM0021622415, plan (intention): Name: Molekulární podstata buněčných a tkáňových regulací

Investor: Ministry of Education, Youth and Sports of the CR, Molecular basis of cell and tissue regulations