ABSTRACTS – ORAL PRESENTATIONS - AMCA, spol. s r.o.

66. DNA REPAIR STUDIES IN LIVING CELLS BY THE USE OF CONFOCAL MICROSCOPY
Eva Bártová, Soňa Legartová, Petra Sehnalová, Lenka Stixová, Stanislav Kozubek
Institute of Biophysics, Academy of Science of Czech Republic, v.v.i., Brno, Czech Republic;
bartova@ibp.cz
The maintenance of genome integrity is fundamental for proper cellular functions.
Genomes are continuously exposed to genotoxic injuries, including UV irradiation
and oxidative stress caused by pollutants. Thus, starting an appropriate DNA repair
signaling pathway is more than demanding for genome stability. Genotoxic stress
generally leads to induction of strand breaks in DNA and especially double-stranded
breaks are dangerous in terms of their faulty repair. The result of inappropriate DNA
repair is the emergence of mutations or chromosomal translocations leading to cancer
progression. Thus, here, we tried to address the function of epigenetic factors, including
histone acetylation, phosphorylation and methylation during DNA damage response. In
addition, in living cells, we analyzed histone code-related proteins and their functional
properties in relationship to optimal DNA repair. In tumor cells, we found acetylationdependent
recruitment of BMI1 and HP1β proteins into locally induced DNA lesions.
Moreover, appearance of Polycomb group-related BMI1 protein and heterochromatin
protein HP1β to DNA lesions was ATPdependent. Changes in histone acetylation and
phosphorylation of H2AX, we also studied in embryonic stem cells, characterized by
acetylation-dependent recruitment of OCT4 protein to DNA lesions, induced in living
cells by local micro-irradiation. Besides OCT4, transcription factors UBFs were recruited
to DNA lesions induced in both nucleoli and non-nucleolar compartment of interphase
nuclei. These experiments showed us a new direction of how to study the fate of
transcription factors after micro-irradiation of living cells.
Acknowledgements
Work was supported by Grant Agency of the Czech Republic, project No.: 13-07822S
and by national COST-CZ project No.: LD11020.
67. INTRODUCTION TO AUTOMATED MICROSCOPY FOR HIGH-THROUGHPUT AND
HIGH-CONTENT SCREENING
Eva Vesela 1 , Tomas Furst 2 , Lenka Radova 3 and Martin Mistrik 1
1
Laboratory of Integrity of Genome, Institute of Molecular and Translational Medicine,
Palacky University, Olomouc, Czech Republic; eva.vesela@upol.cz
2
Department of Matematical Analysis and Math Apllications, Faculty of Science, Palacky
University,Olomouc, Czech Republic
3
Laboratory of Experimental Medicine, Institute of Molecular and Translational
Medicine, Palacky University, Olomouc, Czech Republic
Microscopy-based readout provides the most detailed single cell evaluation. Recent
Analytical Cytometry VII 83