Ekspresija gena uključenih u popravak križnih veza DNA i proteina u zebricama (Danio rerio) koje nose mutaciju u genu tirozil-DNA fosfodiesteraza 2b

master's thesis

• Title (english): Expression of genes involved in DNA-protein crosslink repair in zebrafish (Danio rerio) carrying a mutation in the tyrosyl-DNA phosphodiesterase 2b gene
• Author: Petra Radović
• Mentor: Cecile Francoise Eleonore Otten (mentor)
• Mentor: Ivana Ivančić Baće (komentor)
• Committee member: Ivana Ivančić Baće (predsjednik povjerenstva)
• Committee member: Draginja Mrvoš-Sermek (član povjerenstva)
• Committee member: Mirela Sertić Perić (član povjerenstva)
• Committee member: Mirta Rubčić (član povjerenstva)
• Granter: University of Zagreb; Faculty of Science; (Department of Biology); Zagreb
• Defense date and country: 2024-09-17, Croatia
• Scientific / art field, discipline and subdiscipline: NATURAL SCIENCES; Biology

Abstract

Križne veze DNA i proteina (DPC) su oštećenja DNA u kojima su proteini nepovratno križno
vezani s DNA. Zbog njihove veličine i kemijske stabilnosti, potrebno ih je ukloniti kako bi se
omogućio nastavak bitnih procesa, uključujući replikaciju i transkripciju. Specifični put
popravka nazvan DPC popravkom (DPCR) nedavno je otkriven, a način djelovanja nekoliko
ključnih enzima proučavan je u uvjetima in vitro i u staničnoj kulturi. Jedan takav enzim je
tirozil-DNA fosfodiesteraza 2 (Tdp2), koja sudjeluje u uklanjanju umrežene topoizomeraze 2
iz DNA. Osim proteaza i nukleaza, za potpuni popravak DPC oštećenja vjerojatno su potrebni
i drugi pomoćni proteini. Za proučavanje potencijalnih interakcija između produkta gena tdp2
i popravka DPCR in vivo, upotrijebila sam zebricu Danio rerio, koja je dobro okarakteriziran
mali kralježnjak koji se rutinski koristi kao životinjski model. U ovom radu istražila sam
promjene ekspresije gena u mutantu tdp2b zebrice, dominantnog ortologa odgovornog za
većinu aktivnosti Tdp2 u embrijima zebrice. Od svih analiziranih gena, analiza ekspresije
otkrila je značajno smanjenu ekspresiju znf451 i mre11a, te pojačanu regulaciju gena acrc što
bi moglo ukazivati na potencijalnu međuovisnost s genom tdp2b kod zebrica.

Abstract (english)

DNA-Protein Crosslinks (DPCs) are DNA lesions in which proteins are irreversibly crosslinked
to DNA. Due to their size and chemical stability, they need to be removed to enable
essential processes including replication and transcription to proceed. A specific repair pathway
termed DPC repair (DPCR) has recently been uncovered and the mode of action of several key
enzymes has been studied in vitro and in cell culture. One such enzyme is the tyrosyl-DNA
phosphodiesterase 2 (Tdp2), which is involved in the removal of cross-linked Topoisomerase
2 from DNA. However, proteases, nucleases and other accessory proteins are likely required
for the complete repair of DPC lesions. To study the potential interactions between tdp2 gene
product and DPCR in vivo, I used the zebrafish Danio rerio, which is a well-characterized small
vertebrate that is routinely used as an animal model. In this work, I investigated gene expression
changes in the tdp2b zebrafish mutant, of the more dominantly expressed orthologue that
accounts for the majority of Tdp2 activity in zebrafish embryos. Out of all analysed genes,
expression analysis revealed significantly reduced expression of znf451 and mre11a, and
upregulation of acrc gene that could indicate potential interdependency with tdp2b gene in
zebrafish.

• Keywords: popravak DNA; DPCR; genska promjena
• Keywords (english): DNA repair; DPCR; gene modification
• Language: english
• URN:NBN: urn:nbn:hr:217:777029
• Study programme: Title: Integrated undergraduate and graduate university study of Biology and Chemistry Education; specializations in: Integrated undergraduate and graduate university study of Biology and Chemistry Education; Course: Integrated undergraduate and graduate university study of Biology and Chemistry Education; Study programme type: university; Study level: integrated undergraduate and graduate; Academic / professional title: magistar/magistra edukacije biologije i kemije (magistar/magistra edukacije biologije i kemije)
• Type of resource: Text
• File origin: Born digital
• Access conditions: Open access
• Created on: 2024-09-22 11:03:46