Jana Krejčí

Czech Academy of Sciences, Institute of Biophysics

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Eva Bártová

Czech Academy of Sciences

Stanislav Kozubek

Czech Academy of Sciences, Institute of Biophysics

Soňa Legartová

Czech Academy of Sciences, Institute of Biophysics

Andrea Harničarová

Czech Academy of Sciences

Roman Hájek

University Hospital Ostrava

Gabriela Galiová

Czech Academy of Sciences

Hana Uhlířová

Czech Academy of Sciences, Institute of Scientific Instruments

Orazio Angelo Arcidiacono

Masaryk University

Tomáš Pikálek

Czech Academy of Sciences, Institute of Scientific Instruments

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Tomáš Čižmár

Czech Academy of Sciences, Institute of Scientific Instruments

Cooperative Institutions

Czech Academy of Sciences

Masaryk University

Czech Academy of Sciences, Institute of Biophysics

Charles University

University Hospital Brno

Central European Institute of Technology

Central European Institute of Technology – Masaryk University

Czech Academy of Sciences, Institute of Scientific Instruments

St. Anne's University Hospital Brno

Palacký University Olomouc

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110 μm thin endo-microscope for deep-brain in vivo observations of neuronal connectivity, activity and blood flow dynamics

Zenodo (CERN European Organization for Nuclear Research) (2023)

Miroslav Stibůrek Petra Ondráčková Tereza Tučková Sergey Turtaev Martin Šiler

Dynamics

10.5281/zenodo.7940322

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Post‐Translational Modifications of Histones in Human Sperm

Journal of Cellular Biochemistry (2015)

Jana Krejčí Lenka Stixová Eva Pagáčová Soňa Legartová Stanislav Kozubek

ABSTRACT We examined the levels and distribution of post‐translationally modified histones and protamines in human sperm. Using western blot immunoassay, immunofluorescence, mass spectrometry (MS), and FLIM‐FRET approaches, we analyzed the status of histone modifications and the protamine P2. Among individual samples, we observed variability in the levels of H3K9me1, H3K9me2, H3K27me3, H3K36me3, and H3K79me1, but the level of acetylated (ac) histones H4 was relatively stable in the sperm head fractions, as demonstrated by western blot analysis. Sperm heads with lower levels of P2 exhibited lower levels of H3K9ac, H3K9me1, H3K27me3, H3K36me3, and H3K79me1. A very strong correlation was observed between the levels of P2 and H3K9me2. FLIM‐FRET analysis additionally revealed that acetylated histones H4 are not only parts of sperm chromatin but also appear in a non‐integrated form. Intriguingly, H4ac and H3K27me3 were detected in sperm tail fractions via western blot analysis. An appearance of specific histone H3 and H4 acetylation and H3 methylation in sperm tail fractions was also confirmed by both LC‐MS/MS and MALDI‐TOF MS analysis. Taken together, these data indicate that particular post‐translational modifications of histones are uniquely distributed in human sperm, and this distribution varies among individuals and among the sperm of a single individual. J. Cell. Biochem. 116: 2195–2209, 2015. © 2015 Wiley Periodicals, Inc.

Histone H4

10.1002/jcb.25170

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Citations (30)

Irradiation by γ-rays reduces the level of H3S10 phosphorylation and weakens the G2 phase-dependent interaction between H3S10 phosphorylation and γH2AX

Biopolymers and Cell (2019)

Eva Bártová Gabriela Lochmanová Soňa Legartová Jana Suchánková Radek Fedr

10.7124/bc.0009eb

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Single-cell c-myc gene expression in relationship to nuclear domains

Chromosome Research (2008)

Eva Bártová Andrea Harničarová Jana Krejčí Luděk Strašák Stanislav Kozubek

Nucleoplasm

Transcription

Interphase

10.1007/s10577-007-1196-0

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Citations (11)

[Primary pulmonary adenocarcinoma (pulmonary adenocarcinoma of the alveolar type)].

PubMed (1958)

C Simeĉek Jana Krejčí B Simecková

Pulmonary adenocarcinoma

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Cell differentiation and aging accompanied by depletion of the ACE2 protein

Aging (2020)

Eva Bártová Soňa Legartová Jana Krejčí Orazio Angelo Arcidiacono

ACE2 was observed as the cell surface receptor of the SARS-CoV-2 virus. Interestingly, we also found ACE2 positivity inside the cell nucleus. The ACE2 levels changed during cell differentiation and aging and varied in distinct cell types. We observed ACE2 depletion in the aortas of aging female mice, similarly, the aging caused ACE2 decrease in the kidneys. Compared with that in the heart, brain and kidneys, the ACE2 level was the lowest in the mouse lungs. In mice exposed to nicotine, ACE2 was not changed in olfactory bulbs but in the lungs, ACE2 was upregulated in females and downregulated in males. These observations indicate the distinct gender-dependent properties of ACE2. Differentiation into enterocytes, and cardiomyocytes, caused ACE2 depletion. The cardiomyogenesis was accompanied by renin upregulation, delayed in HDAC1-depleted cells. In contrast, vitamin D2 decreased the renin level while ACE2 was upregulated. Together, the ACE2 level is high in non-differentiated cells. This protein is more abundant in the tissues of mouse embryos and young mice in comparison with older animals. Mostly, downregulation of ACE2 is accompanied by renin upregulation. Thus, the pathophysiology of COVID-19 disease should be further studied not only by considering the ACE2 level but also the whole renin-angiotensin system.

10.18632/aging.202221

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Cell Differentiation and Aging Lead To Up-Regulation of FTO, While the ALKBH5 Protein Level Was Stable During Aging but Up-Regulated During in vitro-Induced Cardiomyogenesis

Physiological Research (2023)

Jana Krejčí OA Arcidiacono Radim Čegan Katarzyna Anna Radaszkiewicz Jiřı́ Pachernı́k

FTO and ALKBH5 proteins are essential erasers of N6-adenosine methylation in RNA. We studied how levels of FTO and ALKBH5 proteins changed during mouse embryonic development, aging, cardiomyogenesis, and neuroectodermal differentiation. We observed that aging in male and female mice was associated with FTO up-regulation in mouse hearts, brains, lungs, and kidneys, while the ALKBH5 level remained stable. FTO and ALKBH5 proteins were up-regulated during experimentally induced cardiomyogenesis, but the level of ALKBH5 protein was not changed when neuroectodermal differentiation was induced. HDAC1 depletion in mouse ES cells caused FTO down-regulation. In these cells, mRNA, carrying information from genes that regulate histone signature, RNA processing, and cell differentiation, was characterized by a reduced level of N6-adenosine methylation in specific gene loci, primarily regulating cell differentiation into neuroectoderm. Together, when we compared both RNA demethylating proteins, the FTO protein level undergoes the most significant changes during cell differentiation and aging. Thus, we conclude that during aging and neuronal differentiation, m6A RNA demethylation is likely regulated by the FTO protein but not via the function of ALKBH5.

RNA methylation

10.33549/physiolres.935078

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Citations (1)

Function of heterochromatin protein 1 during DNA repair

PROTOPLASMA (2017)

Eva Bártová Barbora Malyšková Denisa Komůrková Soňa Legartová Jana Suchánková

Heterochromatin protein 1

HMG-box

Replication protein A

10.1007/s00709-017-1090-3

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Citations (21)

Deacetylation of Histone H4 Accompanying Cardiomyogenesis is Weakened in HDAC1-Depleted ES Cells

International Journal of Molecular Sciences (2018)

Orazio Angelo Arcidiacono Jana Krejčí Jana Suchánková Eva Bártová

Cell differentiation into cardiomyocytes requires activation of differentiation-specific genes and epigenetic factors that contribute to these physiological processes. This study is focused on the in vitro differentiation of mouse embryonic stem cells (mESCs) induced into cardiomyocytes. The effects of clinically promising inhibitors of histone deacetylases (HDACi) on mESC cardiomyogenesis and on explanted embryonic hearts were also analyzed. HDAC1 depletion caused early beating of cardiomyocytes compared with those of the wild-type (wt) counterpart. Moreover, the adherence of embryonic bodies (EBs) was reduced in HDAC1 double knockout (dn) mESCs. The most important finding was differentiation-specific H4 deacetylation observed during cardiomyocyte differentiation of wt mESCs, while H4 deacetylation was weakened in HDAC1-depleted cells induced to the cardiac pathway. Analysis of the effect of HDACi showed that Trichostatin A (TSA) is a strong hyperacetylating agent, especially in wt mESCs, but only SAHA reduced the size of the beating areas in EBs that originated from HDAC1 dn mESCs. Additionally, explanted embryonic hearts (e15) responded to treatment with HDACi: all of the tested HDACi (TSA, SAHA, VPA) increased the levels of H3K9ac, H4ac, H4K20ac, and pan-acetylated lysines in embryonic hearts. This observation shows that explanted tissue can be maintained in a hyperacetylation state several hours after excision, which appears to be useful information from the view of transplantation strategy and the maintenance of gene upregulation via acetylation in tissue intended for transplantation.

Trichostatin A

HDAC1

10.3390/ijms19082425

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Citations (10)

Advanced Confocal Microscopy Techniques to Study Protein-protein Interactions and Kinetics at DNA Lesions

Journal of Visualized Experiments (2017)

Soňa Legartová Jana Suchánková Jana Krejčí Alena Svobodová Kovaříková Eva Bártová

Local microirradiation with lasers represents a useful tool for studies of DNA-repair-related processes in live cells. Here, we describe a methodological approach to analyzing protein kinetics at DNA lesions over time or protein-protein interactions on locally microirradiated chromatin. We also show how to recognize individual phases of the cell cycle using the Fucci cellular system to study cell-cycle-dependent protein kinetics at DNA lesions. A methodological description of the use of two UV lasers (355 nm and 405 nm) to induce different types of DNA damage is also presented. Only the cells microirradiated by the 405-nm diode laser proceeded through mitosis normally and were devoid of cyclobutane pyrimidine dimers (CPDs). We also show how microirradiated cells can be fixed at a given time point to perform immunodetection of the endogenous proteins of interest. For the DNA repair studies, we additionally describe the use of biophysical methods including FRAP (Fluorescence Recovery After Photobleaching) and FLIM (Fluorescence Lifetime Imaging Microscopy) in cells with spontaneously occurring DNA damage foci. We also show an application of FLIM-FRET (Fluorescence Resonance Energy Transfer) in experimental studies of protein-protein interactions.

Photobleaching

Bimolecular fluorescence complementation

Fluorescence-lifetime imaging microscopy

10.3791/55999

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Citations (7)