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Analysis of endomembrane markers in the cortical cytoplasm and their co-localization with Arp2/3 complex
Jelínková, Barbora ; Schwarzerová, Kateřina (advisor) ; Fendrych, Matyáš (referee)
ARP2/3 is an evolutionarily conserved heteroheptameric protein complex. Its main activity lies in the nucleation of dendritic actin filaments that are involved in membrane remodeling. ARP2/3 takes part in plasma membrane remodeling and the formation of cytoplasmic protrusions that serve in the amoeboid motion of mammalian cells and some protists and plays role in exocytosis and endocytosis of animal and yeast cells. The main objective of this work was to find a connection between the ARP2/3 complex and the regulation of the plant endomembrane system. Using TIRF microscopy we visualized the localization of the ARP2/3 complex in the cortical layer of plant cells and compared it to the localization of several endomembrane markers from the Rab family and an exocytotic marker Exo84b. In the vicinity of the plasma membrane, the ARP2/3 complex subunits localized to dynamic dots very similar to the localization of Exo84b protein. Colocalization analysis showed that a small portion of Exo84b marker and ARP2/3 complex signals colocalize and this result was seconded by the biochemical approach of coimmunoprecipitation. Key words: ARP2/3, endomembrane system, cortical layer, RabA1g, RabC1, RaD2a, Exo84b
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Structure- and sequence-based identification of functionally important amino acids in a protein family
Peclinovská, Iveta ; Novotný, Marian (advisor) ; Pleskot, Roman (referee)
A group of small GTPases consist of over twenty protein families in the super class P-loop. It has a very diverse cell functions. Small GTPases regulate the formation of vesicular follicles, cytoskeleton and nuclear transport. They participate also on cell proliferation and signaling. The aim of my work is to find important amino acids that define family and distinguish each other. I focus on families Arf, Rab, Ran, Ras and Rho. At the Rho family I am also devoted to classes Rho, Rac and Cdc42. Amino acids are identified using bioinformatic programs selected Consurf and Sca5. The objective is also to test P2RANK specialized tool developed at the Charles University in Prague that predict ligand binding sites from protein structure in different families. Founding amino acids can have a big role in the functional divergence of individual families and classes of small GTPases and can be the basis for future study example for the proliferation of cancerous cells. 1.1 Keywords Powered by TCPDF (www.tcpdf.org)
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Vesicular trafficking into the plant vacuole
Semerádová, Hana ; Kulich, Ivan (advisor) ; Vosolsobě, Stanislav (referee)
Vacuole is very important plant cell organelle which can occupy 90% of cell volume. It provides wide range of functions. Considering enormous size of the vacuole, vesicle trafficking into the plant vacuole is major vesicle movement inside the cell. Transport pathway into the vacuole is very dynamic field of plant cell biology. This sorting machinery shares similarities within all eukaryotes, but plants also have their own specificities. Soluble cargo is targeted through secretory pathways by vacuolar sorting receptors (VSRs). These proteins due to its transmembrane localization can interact with sorted cargo and take it to the right organelle within the cell. Membrane fusion is facilitated with Rab-GTPase and SNARE protein complexes. A special kind of vesicle traffic is autophagy, the self consuming process that protects the cell from various type of stress or enables apoptosis.
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