Določanje proteinov, vezanih na dipeptidne ponovitve

Markič, Lara (author), Rogelj, Boris (mentor)

Celični mehanizmi, odgovorni za nastanek patologije, ki vodi v amiotrofično lateralno sklerozo, še niso dobro poznani in trenutno ni zdravil, ki bi bolezen pozdravila. Proteini z dipeptidnimi ponovitvami (DPR), ki se prevajajo iz podaljšanih heksanukleotidnih ponovitev znotraj gena C9orf72, imajo sposobnost vezave na različne znotrajcelične proteine. Le-ti pri bolnikih z amiotrofično lateralno sklerozo nastanejo pri posebni obliki translacije, ki poteka neodvisno od začetnega kodona AUG. V nalogi smo izražali pet različnih zvrsti dipeptidnih ponovitev (poli (GA), poli (GP), poli (GR), poli (PA) in poli (PR)), ki smo jih konjugirali z encimom BioID2, in z metodo BioID poiskali kandidatne celične proteine, ki se prehodno ali stalno vežejo na DPR, ali pa se nahajajo v njihovi bližini. Z ontološkimi analizami kandidatnih proteinov smo določili, da so proteini poli (GA) vpleteni v procese, povezane z mikrotubuli, in v katabolične procese proteinov; proteini poli (GP) vpleteni v procese prenosa signala; proteini poli (GR) vpleteni v procese začetka prevajanja proteinov in metabolične procese amidov; proteini poli (PA) vpleteni v uravnavanje encimske aktivnosti in pozitivnega uravnavanja apoptoze; proteini poli (PR), vpleteni v procesiranje rRNA. Z magistrsko nalogo smo prispevali nova spoznanja o celičnih mehanizmih pri najpogostejši mutaciji amiotrofične lateralne skleroze, ki bodo, skupaj z nadaljnjimi raziskavami, pripomogla k boljšemu razumevanju bolezni in novim načinom zdravljenja ali preprečevanja bolezni.

• Language: Slovenian
• Keywords: biotehnologija, nevrodegeneracija, proteini, interaktorji, amiotrofična, lateralna, skleroza, frontotemporalna, demenca, C9orf72, C9-ALS, ALS, dipeptidne, ponovitve, RAN-translacija
• Work type: Master's thesis/paper
• Typology: 2.09 - Master's Thesis
• Organization: BF - Biotechnical Faculty
• Publisher: [L. Markič]
• Year: 2019
• PID: 20.500.12556/RUL-107622
• UDC: 606:616.8-009.5:577.1:576.54(043.2)
• COBISS.SI-ID: 9215609
• Publication date in RUL: 07.05.2019

Secondary language

• Language: English
• Title: Determination of proteins bound to dipeptide repeats

Abstract

The cellular mechanisms that are responsible for the development of pathology leading to amyotrophic lateral sclerosis are not yet well understood, and currently there is no cure for the disease. Dipeptide repeat (DPR) proteins that are translated from extended hexanucleotide repeats within the C9orf72 gene can bind to various intracellular proteins. These proteins in patients with amyotrophic lateral sclerosis are generated by a special form of repeat-associated non-AUG translation. We expressed five different types of dipeptide repeats (poly(GA), poly(GP), poly(GR), poly(PA) and poly(PR)), which were conjugated with the BioID2 enzyme, and with the BioID method determined candidate intracellular proteins that transiently or permanently bind to DPRs or can be found in their vicinity. With ontological analysis of candidate proteins we determined that poly(GA) proteins are involved in microtubule-based processes and in protein catabolic processes; poly(GP) proteins are involved in signal transduction processes; poly(GR) proteins are involved in translational initiation processes and in cellular amide processes; poly (PA) proteins are involved in the regulation of catalytic activity and in positive regulation of apoptotic processes; poly (PR) proteins are involved in the rRNA processing. With this master thesis we have contributed new findings about cellular mechanisms in the most common mutation of amyotrophic lateral sclerosis, which will along with further research contribute to a better understanding of the disease as well as new ways of treatment and prevention of the disease.

• Keywords: biotechnology, neurodegeneration, proteins, interactors, amyotrophic, lateral, sclerosis, frontotemporal, dementia, C9orf72, C9-ALS, ALS, dipeptide, repeats, RAN-translation