Experimental systems for identification of proteins involved in stability of palindromes in DNA

Palindromske sekvencije predstavljaju posebnu skupinu obrnutih ponavljanja koja nisu odijeljena niti jednim parom baza, a pronađene su u genomima svih organizama. Iako imaju važnu funkcionalnu ulogu u regulatornim dijelovima genoma kao što su promotori, terminatori i ishodišta replikacije, duži palindromi predstavljaju opasnost za stabilnost genoma budući da mogu tvoriti sekundarne strukture koje uzrokuju pucanje kromosoma te preraspodjelu genetičkog materijala, što može dovesti i do teških genetičkih bolesti kod ljudi. Pretpostavlja se da je nestabilnost palindroma posredovana cijelim nizom proteina, od kojih mnogi još uvijek nisu poznati. Zbog toga je cilj ovoga rada bio konstruirati eksperimentalni sustav za brzu i jednostavnu identifikaciju tih proteina, kako u genomu kvasca, tako i u humanom genomu. Primjena ovog sustava za identifikaciju nepoznatih proteina uključenih u (ne)stabilnost palindroma mogla bi unaprijediti buduća istraživanja genetičkih poremećaja potaknutih postojanjem palindroma u molekuli DNA.Palindromic sequences represent a special group of inverted repeats that are not seperated by any base pairs, and are found in genomes of all organisms. Although they have important functional role in regulatory regions of genome such as promotors, terminators and replication initiation sites, longer palindromes represent danger for the genome stability due to their ability to form secondary structures that act as hotspots for chromosome breakage and genomic rearrangements, which can lead to severe genetic diseases in humans. It is believed that palindrome instability is mediated by an array of various proteins, many of which are still unknown. Therefore, the purpose of this thesis was developing an experimental system for quick and simple identification of those proteins in yeast, as well as in human genome. The usage of this system for identification of unknown proteins involved in palindrome (in)stability could improve future research of palindrome-mediated genetic disorders in DNA molecule