|
|
Genetically encoded biosensors of cellular tension and their use in cellular biology
Pelantová, Markéta ; Rösel, Daniel (advisor) ; Lánský, Zdeněk (referee)
1 Abstract and key words Mechanical forces have great impact on the life of cells. They influence cell proliferation, migration or differentiation and defects in cellular mechanosensing were reported to be the cause of various diseases, such as deafness, atherosclerosis or cancer. However, mechanisms of mechanical sensing are not thoroughly examined and not many tools for doing such research are available. Genetically encoded FRET-based biosensors are one of the existing methods for studying transfer of mechanical signal in cells. It is a non-invasive method allowing to observe changes in mechanical tension across proteins in living cells. In this thesis, different types of existing genetically encoded FRET-based tension biosensors are introduced together with the process of their development and knowledge gained by their use in research. Key words: mechanical force, mechanosensing, FRET, tension sensor, biosensor development
|
|
|
Molecular mechanism of microtubule severing by katanin
Podhájecký, Roman ; Lánský, Zdeněk (advisor) ; Libusová, Lenka (referee)
Microtubule cytoskeleton is a dynamic filamentous network, whose reorga- nization underpins important cellular processes, such as cell motility or cell di- vision. This remodeling highly depends on microtubule-associated proteins that can remodel individual microtubules, such as katanin. Katanin is a microtubule- associated protein that employs ATP hydrolysis to sever microtubules. This function can rapidly reorganize microtubule networks by providing fast amplification of the microtubule polymer number as well as microtubule fast degradation. The exact mechanism of microtubule severing and its regulation by katanin is largely dis- cussed nowadays. In this project I would like to describe katanin's mechanism of action and its regulation on a molecular level. Key words: cytoskeleton, remodelling of microtubule networks, microtubules, microtubule se- vering enzymes, katanin 1
|
|
|
Characterization of WASH complex member protein SWIP
Humhalová, Tereza ; Libusová, Lenka (advisor) ; Lánský, Zdeněk (referee)
WASH complex regulates actin dynamics on endosomes by activating the Arp2/3 complex, which subsequently induces generation of branched actin patches. WASH complex is required for proper recycling of many important transmembrane proteins. Although the general physiological function of WASH complex is known, the role of its single subunits have not yet been adequately specified. This work focuses on one of these subunits - protein SWIP. This protein maintains vesicular localization of some WASH complex subunits in the slime mold Dictyostelium discoideum and a point mutation in its sequence causes a severe neurodegenerative disease - autosomal recessive intellectual disorder (ARID). Our results show that SWIP truncation results in its inability to incorporate into WASH complex. However, the C-terminal part of SWIP is able to localize onto intracellular vesicles, which are not WASH complex positive. We have also studied the impact of ARID-causing SWIP mutation, and we show, that it does neither change the protein's ability to bind the complex nor the overall localization of WASH complex.
|
|
| |
|
| |
guest ::
