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master's thesis
Uloga proteina ACRC u popravku unakrsnih veza DNA-protein u modelnom organizmu zebrici (Danio rerio)
Zagreb: University of Zagreb, Faculty of Science, 2021. urn:nbn:hr:217:305599
Kutnjak, Marin
University of Zagreb
Faculty of Science
Department of Biology

Institutional repository: Repository of the Faculty of Science

Cite this document

Kutnjak, M. (2021). Uloga proteina ACRC u popravku unakrsnih veza DNA-protein u modelnom organizmu zebrici (Danio rerio) (Master's thesis). Zagreb: University of Zagreb, Faculty of Science. Retrieved from https://urn.nsk.hr/urn:nbn:hr:217:305599

Kutnjak, Marin. "Uloga proteina ACRC u popravku unakrsnih veza DNA-protein u modelnom organizmu zebrici (Danio rerio)." Master's thesis, University of Zagreb, Faculty of Science, 2021. https://urn.nsk.hr/urn:nbn:hr:217:305599

Kutnjak, Marin. "Uloga proteina ACRC u popravku unakrsnih veza DNA-protein u modelnom organizmu zebrici (Danio rerio)." Master's thesis, University of Zagreb, Faculty of Science, 2021. https://urn.nsk.hr/urn:nbn:hr:217:305599

Kutnjak, M. (2021). 'Uloga proteina ACRC u popravku unakrsnih veza DNA-protein u modelnom organizmu zebrici (Danio rerio)', Master's thesis, University of Zagreb, Faculty of Science, accessed 22 November 2024, https://urn.nsk.hr/urn:nbn:hr:217:305599

Kutnjak M. Uloga proteina ACRC u popravku unakrsnih veza DNA-protein u modelnom organizmu zebrici (Danio rerio) [Master's thesis]. Zagreb: University of Zagreb, Faculty of Science; 2021 [cited 2024 November 22] Available at: https://urn.nsk.hr/urn:nbn:hr:217:305599

M. Kutnjak, "Uloga proteina ACRC u popravku unakrsnih veza DNA-protein u modelnom organizmu zebrici (Danio rerio)", Master's thesis, University of Zagreb, Faculty of Science, Zagreb, 2021. Available at: https://urn.nsk.hr/urn:nbn:hr:217:305599

Metadata
TitleUloga proteina ACRC u popravku unakrsnih veza DNA-protein u modelnom organizmu zebrici (Danio rerio)
Title (english)The role of ACRC protein in the DNA-protein crosslink repair in zebrafish (Danio rerio) model organism
AuthorMarin Kutnjak
MentorMarta Popović (mentor)
MentorNenad Malenica (sumentor)
Committee memberNenad Malenica (predsjednik povjerenstva)
Committee memberŽeljka Vidaković-Cifrek (član povjerenstva)
Committee memberMarin Ježić (član povjerenstva)
Committee memberIvana Ivančić Baće (član povjerenstva)
GranterUniversity of Zagreb
Faculty of Science
(Department of Biology)
Zagreb
Defense date and country2021-09-30, Croatia
Scientific / art field,
discipline and subdiscipline		NATURAL SCIENCES
Biology
Abstract
Proteini koji stupaju u interakciju s DNA ili se nalaze u njenoj blizini mogu postati s njom ireverzibilno kovalentno vezani, tvoreći lezije nazvane unakrsne veze DNA-protein (DPC-ovi, engl. DNA-protein crosslinks). DPC-ovi ometaju stanične procese koji uključuju DNA (replikaciju, transkripciju, rekombinaciju, popravak DNA), stoga predstavljaju prepreku u održavanju genomskog integriteta. Protein SPRTN prva je otkrivena proteaza uključena u popravak DPC-ova u metazoa. Protein ACRC/GCNA evolucijski je očuvan regulator genomske stabilnosti metazoa. Eksprimiran je u germinativnim i pluripotentnim stanicama. Modeliranjem in silico uočena je strukturna sličnost proteaznih jezgara proteina SPRTN i ACRC. Stanice s mutiranim genom acrc karakteriziraju nestabilnost kromosoma, replikacijski stres i nakupljanje DPC-ova. Tijekom izvedbe diplomskog rada sudjelovao sam u uzgoju mutanata zebrice (Danio rerio) za protein ACRC kako bih istražio njegovu ulogu u popravku DPC-ova. Određivao sam razinu ukupnih DPC-ova embrija zebrice s delecijom C-kraja proteina i delecijom pretpostavljenog katalitičkog glutamata E451. Također sam izvodio ciljanu inaktivaciju gena Acrc u embrijima zebrice metodom CRISPR/Cas. Rezultati rada pokazali su da bi E451 i C-kraj proteina ACRC zebrice mogli biti bitni za njegovu katalitičku aktivnost u popravku DPC-ova, te da je specifičnim RNA vodiljama (gRNA) za 5'-kraj i 3'-kraj gena Acrc zebrice moguće djelomično ciljano inaktivirati ovaj gen metodom CRISPR/Cas.
Abstract (english)
Proteins which interact with DNA or are in its proximity can get irreversibly covalently bonded with DNA and form lesions called DNA-protein crosslinks (DPCs). DPCs impair cellular processes which include DNA and are obstacles in genomic stability maintenance. SPRTN protein was a first discovered protease involved in DPC repair in metazoans. ACRC/GCNA protein is a conserved regulator of genomic stability of metazoans. It’s expressed in germ and pluripotent cells. Structural modelling of SPRTN and ACRC protease cores revealed similarities between them. Cells with mutated acrc gene are prone to chromosome instability, replication stress and DPCs accumulation. In my research, I participated in creation of zebrafish (Danio rerio) ACRC mutants, with the goal to investigate its role in DPC repair. I have determined total DPC levels of zebrafish embryos with deletion of C-end and deletion of putative catalytic glutamate E451 of ACRC protein. I also performed targeted gene inactivation of Acrc gene in zebrafish embryos with CRISPR/Cas method. My results showed that E451 and C-end of ACRC protein could be important for its catalytic activity in DPC repair, and that it’s possible to partially inactivate Acrc gene with CRISPR/Cas method using specific guide RNAs for 5’- and 3’-end of this gene.
Keywords
Keywords (english)
Languagecroatian
URN:NBNurn:nbn:hr:217:305599
Study programmeTitle: Molecular Biology
Study programme type: university
Study level: graduate
Academic / professional title: magistar/magistra molekularne biologije (magistar/magistra molekularne biologije)
Type of resourceText
File originBorn digital
Access conditionsOpen access
Embargo expiration date: 2023-09-30
Terms of useLicence In copyright icon
Created on2021-10-08 11:23:27