Ekspresija gena xpa, acrc i tdp1 u ribe zebrice (Danio rerio) i identifikacija zebrica mutiranog gena xpa

master's thesis

• Title (english): Expression of xpa, acrc and tdp1 genes in zebrafish (Danio rerio) and identification of xpa deficient mutants
• Author: Luka Vinković
• Mentor: Marta Popović (mentor)
• Mentor: Nenad Malenica (komentor)
• Committee member: Nenad Malenica (predsjednik povjerenstva)
• Committee member: Inga Urlić (član povjerenstva)
• Committee member: Ivana Ivančić Baće (član povjerenstva)
• Committee member: Maja Matulić (član povjerenstva)
• Granter: University of Zagreb; Faculty of Science; (Department of Biology); Zagreb
• Defense date and country: 2022-12-06, Croatia
• Scientific / art field, discipline and subdiscipline: NATURAL SCIENCES; Biology

Abstract

Križne veze DNA i proteina su ireverzibilne, kovalentne veze DNA i proteina koje
ograničavaju obavljanje staničnih procesa. Protein Acrc se zbog metaloproteazne domene i
uloge u očuvanju genomske stabilnosti povezuje s popravkom križnih veza DNA i proteina.
Još jedan mehanizam koji sudjeluje u popravku križnih veza DNA i proteina je popravak
izrezivanjem nukleotida u kojem protein Xpa ima esencijalnu ulogu proteina nosača u ovom
mehanizmu popravka. Tirozil DNA fosfodiesteraza 1 (Tdp1) specifična je za određenu vrstu
oštećenja pa svojom hidrolitičkom aktivnošću razrješuje fosfotirozilnu vezu između
topoizomeraze 1 i jednolančane DNA. Analizom ekspresije gena acrc, xpa i tdp1 lančanom
reakcijom polimeraze u stvarnom vremenu u tkivima ribe zebrice (Danio rerio) uočeno je da,
osim pada ekspresije ovih gena starenjem, postoje tkiva u kojima ekspresija raste, što je
suprotno očekivanjima. Analizom razlike ekspresije gena između spolova uočeno je da je
ekspresija u pojedinim tkivima veća kod ženki zebrice, a u pojedinim kod mužjaka pa se za
određeni spol ne može tvrditi da je otporniji na križne veze DNA i proteina na temelju
ekspresije gena acrc, xpa i tdp1. Uz analizu ekspresije gena cilj je bio identificirati zebrice s
mutiranim genom xpa u svrhu stvaranja Xpa deficijente linije zebrica. Pronađena mutacija nije
inaktivirala protein Xpa.

Abstract (english)

DNA-protein crosslinks are irreversible, covalent bonds of DNA and proteins that limit the cell
performance. Acrc protein is being connected to DNA-protein crosslink repair because of its
metalloprotease domain and a role in genome stability. Another mechanism involved in DNAprotein crosslink repair is nucleotide excision repair in which protein Xpa has an essential
scaffolding role. Tyrosyl DNA phosphodiesterase 1 (Tdp1) is specific for a particular type of
damage as it resolves the phosphotyrosyl bond between topoisomerase I and single-stranded
DNA. Analysis of acrc, xpa and tdp1 gene expression with real-time polymerase chain reaction
in zebrafish (Danio rerio) tissues showed that besides of the reduction in expression of these
genes there are tissues that exhibit increased expression which wasn’t expected. Analysis of
expression differences between zebrafish genders showed that some tissues exhibit higher
expression in females while some have higher expression in males so it can’t be declared that
a particular gender is more resistant to DNA-protein crosslinks based on expression of acrc,
xpa and tdp1 genes. There was also a goal to identify zebrafish with mutated xpa gene with a
purpose of creating a Xpa deficient zebrafish line, but the only detected modification doesn’t
inactivate the protein.

• Keywords: DNA popravak; križne veze DNA i proteina; starenje; ekspresija; genska promjena
• Keywords (english): DNA repair; DNA-protein crosslinks; expression; gene modification
• Language: croatian
• URN:NBN: urn:nbn:hr:217:656517
• Study programme: Title: Molecular Biology; Study programme type: university; Study level: graduate; Academic / professional title: magistar/magistra molekularne biologije (magistar/magistra molekularne biologije)
• Type of resource: Text
• File origin: Born digital
• Access conditions: Embargoed access; Embargo expiration date: 2024-12-06
• Created on: 2022-12-19 12:07:20