Obilježavanje i detekcija proteina Pir3 u različitim mutantima egzosoma kvasca Saccharomyces cerevisiae

undergraduate thesis

• Title (english): Labeling and detection of Pir3 protein in various mutants of the Saccharomyces cerevisiae yeast exosome
• Author: Korina Perišić
• Mentor: Igor Stuparević (mentor)
• Granter: University of Zagreb; Faculty of Food Technology and Biotechnology; (Department of Chemistry and Biochemistry); (Laboratory for Biochemistry); Zagreb
• Defense date and country: 2022-07-18, Croatia
• Scientific / art field, discipline and subdiscipline: BIOTECHNICAL SCIENCES; Interdisciplinary Technical Sciences

Abstract

U eukariotskim stanicama, procesiranje i razgradnja RNA molekula postignuta je aktivnošću multiproteinskog kompleksa RNA egzosoma koji je jedan od važnih regulatora stanične stijenke kvasca. Promjene u staničnoj stijenci događaju se ovisno o fazi staničnog ciklusa, dostupnosti hranjivih tvari i uvjetima okoline kao što su pH, temperatura i dostupnost kisika, a usko su povezane s promjenama u proteinskom sastavu koji se također mijenja kroz različite faze rasta stanice. Jezgra RNA egzosoma sastoji se od devet strukturnih podjedinica na koje su vezane dvije katalitičke podjedinice koje imaju 3'-5' egzoribonukleaznu aktivnost, Dis3 i Rrp6. Za aktivnost RNA egzosoma važnu ulogu ima i prisutnost kofaktora Rrp47, Mpp6 i TRAMP kompleksa. Problemi u funkcioniranju RNA egzosoma utječu na stabilnost stanične stijenke prilikom čega je pogođen i proteinski sastav stijenke. Cilj ovog rada bio je ispitati kako nedostatak pojedinog kofaktora RNA egzosoma utječe na ekspresiju proteina stanične stijenke kvasca, Pir3. U tu svrhu protein Pir3 je obilježen hemaglutininskom oznakom u različitim mutantima kojima nedostaje pojedini kofaktor RNA egzosoma, te je praćena njegova ekspresija tijekom logaritamske i stacionarne faza rasta stanica. Rezultati ovog rada pokazuju kako kofaktori RNA egzosoma imaju utjecaj na ekspresiju proteina stanične stijenke kvasca.

Abstract (english)

In eukaryotic cells, processing and degradation of RNA molecules occurs through the activity of the multiprotein RNA exosome complex, which is one of the major regulators of the yeast cell wall. Changes in the cell wall occur depending on the phase of the cell cycle, the availability of nutrients and environmental conditions such as pH, temperature and oxygen availability, and are closely related to changes in protein composition, which also changes during different phases of cell growth. The core of the RNA exosome consists of nine structural subunits to which two catalytic subunits with 3'-5' exoribonuclease activity, Dis3 and Rrp6, are attached. The presence of the cofactors Rrp47, Mpp6 and the TRAMP complex plays an important role in RNA exosome activity. Problems in the function of RNA exosomes affect the stability of the cell wall, which also affects the protein composition of the wall. The aim of this work was to investigate how the absence of a specific exosome RNA cofactor affects the expression of the yeast cell wall protein, Pir3. To this end, the Pir3 protein was tagged with a hemagglutinin tag in different mutants lacking a specific exosome RNA cofactor and its expression was monitored during the logarithmic and stationary phases of cell growth. The results of this work demonstrate how exosome RNA cofactors affect the expression of yeast cell wall proteins.

• Keywords: kvasac Saccharomyces cerevisiae; stanična stijenka; RNA egzosom; Pir3 protein
• Keywords (english): yeast Saccharomyces cerevisiae; cell wall; RNA exosome; Pir3 protein
• Language: croatian
• URN:NBN: urn:nbn:hr:159:904967
• Study programme: Title: Biotechnology; Study programme type: university; Study level: undergraduate; Academic / professional title: sveučilišni/a prvostupnik/prvostupnica (baccalaureus/baccalaurea) inženjer/inženjerka biotehnologije (sveučilišni/a prvostupnik/prvostupnica (baccalaureus/baccalaurea) inženjer/inženjerka biotehnologije)
• Type of resource: Text
• File origin: Born digital
• Access conditions: Open access
• Created on: 2022-10-21 12:54:35