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Regulation of histone post-translational modifications and its use in the diagnosis and treatment of lung cancer
Marada, Kryštof ; Janoštiak, Radoslav (advisor) ; Červená, Klára (referee)
The aim of this bachelor's thesis is to summarize the epigenetic mechanisms, namely DNA methylation, non-coding RNA and post-translational modification of histones, within the scope of which the five currently most researched types are described, i.e. acetylation, methylation, phosphorylation, ubiquitinylation and sumoylation. Furthermore, a summary of the modifying enzymes whose function is to add and remove these epigenetic components. And also the description of the proteins that recognize these components. The following is a description of how epigenetic processes are involved in the course of cancer. The remaining part of the work is devoted to lung cancer, its most common causes, what methods are currently used to treat it, and how post-translational modifications of histones could be used in its treatment. Most of this chapter is devoted to histone deacetylase inhibitors, as these inhibitors represent potential as a new component of lung cancer treatment, which also makes them the target of much research.
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Post-Translational Modifications of Nuclear and Non-Nuclear Proteins in Spermatozoa
Řimnáčová, Hedvika ; Nevoral, Jan (advisor) ; Krapf, Dario (referee) ; Álvarez Rodríguez, Manuel (referee)
Posttranslational modifications of nuclear and nonnuclear proteins in spermatozoa Summary The number of couples who need the help of assisted reproductive technology (ART) has increased over the years. Approximately half of the cases are caused by male infertility, which is often diagnosed as idiopathic infertility. Therefore, the search for male fertility markers will improve male infertility diagnosis, thereby facilitating advanced sperm treatment and selection via ART. Posttranslational modifications (PTMs) of sperm nuclear and nonnuclear proteins are suitable candidates for such markers. The PTMs of protamines and histones reflect sperm chromatin maturity and its readiness for fertilization, and accordingly, they can predict the outcome of ART. However, the PTMs of nonnuclear proteins, including cytoplasmic, cytoskeletal, and membrane proteins, reflect the ability of sperm to undergo hyperactivation, capacitation, or acrosome reactions, which are processes essential for fertilization. We hypothesize that the PTMs of nuclear and nonnuclear proteins can reflect sperm quality and, thus, serve as a valuable marker in ART. Additionally, we suggest that the in vitro addition of hydrogen sulfide into the sperm-manipulating media improves sperm motility and viability via persulfidation. We used Western blot...
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Chromatin modifications in association with priming phenomenon in plants
Marková, Hana ; Holá, Dana (advisor) ; Fischer, Lukáš (referee)
Plants are exposed to many stressors to which they must be able to adapt. It has been found that plants can memorize the respective stress response and may respond to a subsequent stress exposure faster and to a greater extent. The mechanism of this so-called priming could be associated with the changes in the levels of chromatin modifications. Chromatin modifications are usually dynamically changing during the stress reaction. Some of these changes could persist for some time, thus the specific stress memory, mitotically and meiotically transmissible, could be established. Such a phenomenon was observed for stress caused by dehydration, salinity, mechanical damage, a combination of various stressors and a systemic resistance to pathogenic bacteria. So far, only a few studies on this topic exist; but even now it is clear that there are differences in the response of specific genes as well as the dependence on the length of the stress stimulus and the duration of the period between the first and second stress. A major disadvantage of existing studies is that they focus solely on histone modifications. Regarding the chromatin modifications studied so far, H3K4me3 could serve as the main mark for such priming. On the other hand, H3K27me3 modification is apparently not used as a memory tag. However, it is still...
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Isolation and characterisation of oviductal proteins involved in formation of sperm oviductal reservoir in cow
Kračmerová, Jana ; Liberda, Jiří (advisor) ; Postlerová, Pavla (referee)
The oviduct plays an important role in the complex process of fertilization and in the early phase of embryo development. Isolation and characterization of some components of bovine oviduct participating in the formation of oviductal sperm reservoir was a subject of the present study. Two affinity sorbents containing immobilized components of bull seminal plasma (non-separated proteins of bull seminal plasma and their phosphorylcholine binding fraction) were prepared and used for the study of interaction of immobilized ligands with components of oviductal epithelium. Two types of preparation of oviductal membrane proteins of the isthmus region were isolated: complete membrane proteins and the apical fraction of membrane proteins. These preparations were separated using prepared affinity matrices. in both cases only an insignificant amount of proteins was found in the adsorbed fractions. Further attention has been paid to the identification of four protein zones detected after electrophoretic separation in the presence of SDS (rel. mol. wt. 15 500 - 18 500) in the fraction of apical membrane proteins. These proteins were identified after the electrophoretic separation and tryptic digestion using MS analysis as bovine histones H4, H2A typ 2-C, H2B typ 1-K a H3.3. Proteins identified as bovine...
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