Naslov
The role of GCNA (ACRC) protein in the DNA-protein crosslink repair in zebrafish
(The role of GCNA (ACRC) protein in the DNA-protein crosslink repair in zebrafish)

Autori
Kutnjak, Marin ; Supina Pavić, Christine ; Otten, Cecile ; Popović, Marta

Vrsta, podvrsta i kategorija rada
Sažeci sa skupova, sažetak, znanstveni

Izvornik
Book of Abstracts of the Congress of the Croatian Society of Biochemistry and Molecular Biology "HDBMB22: From Science to Knowledge" / Dulić, Morana ; Sinčić, Nino ; Vrhovac Madunić, Ivana - Zagreb : Hrvatsko Društvo za Biotehnologiju, 2022, 99-99

Skup
International Congress of the Croatian Society of Biochemistry and Molecular Biology "HDBMB22: From Science to Knowledge"

Mjesto i datum
Brela, Hrvatska, 28.09.2022. - 01.10.2022

Vrsta sudjelovanja
Poster

Vrsta recenzije
Međunarodna recenzija

Ključne riječi
DNA repair ; DNA-protein crosslinks (DPCs) ; GCNA (ACRC) protease ; CRISPR/Cas9 ; zebrafish

Sažetak
DNA is a molecule in a chemically reactive environment and under these conditions various DNA lesions are constantly formed in cells. Proteins that interact with DNA or are in its proximity can become irreversibly covalently bound to DNA and form lesions called DNA-protein crosslinks (DPCs). DPCs can be induced by exogenous (UV and ionizing radiation, transition metals, chemotherapeutic agents) and endogenous factors (reactive oxygen and nitrogen species, aldehydes, DNA helix alterations). They affect cellular processes that include DNA (DNA replication, transcription, repair, recombination, and chromatin remodeling) and are obstacles to genome stability maintenance. Several groups have characterized the proteins Wss1 in yeast and SPRTN in metazoans as the first identified proteases involved in DPC repair. At the cellular level, impaired DPC repair can lead to DNA double-strand breaks, genomic instability, and consequently cell death, while at the organismal level it can cause cancer, premature aging, and neurodegenerative diseases. Recent studies have shown that there is a second potential DPC protease in metazoans: GCNA (germ cell nuclear acidic) or ACRC (acidic repeat- containing). Phylogenetic analysis of the SprT and WLM domains and structural modeling of the protease cores of SPRTN and GCNA have revealed similarities between these two proteins. The aim of our study was to determine whether GCNA is involved in DPC repair in vivo and whether its putative protease core is important for GCNA function. We used zebrafish (Danio rerio) as a model organism, CRISPR/Cas9 gene manipulation to generate zebrafish mutant strains, and the RADAR (rapid approach to DNA adduct recovery) method to isolate DPCs from mutant embryos. We introduced a mutation in the putative protease active site of GCNA that includes a deletion of E451 (putative catalytic glutamate). Our results showed that the mutant embryos had 100% mortality within the first 24 hours of development due to maternal effect. Primarily, they had an increased total amount of cellular DPCs. Our study demonstrates the importance of GCNA for embryonic development and DPC repair at the organismal level, as well as the importance of the putative protease core for GCNA function.

Izvorni jezik
Engleski

Znanstvena područja
Biologija

POVEZANOST RADA