Interakcije peptidov domene SUD M Nsp3 virusa SARS-CoV-2 s 3’-neprevedenimi regijami človeške mRNA

Dolenc, Katja (author), Marušič, Maja (mentor), Gunčar, Gregor (comentor)

Koronavirus SARS-CoV-2 je povzročil pandemijo COVID-19, bolezni, ki je v zadnjih letih svojimi številnimi resnimi simptomi pomembno vplivala na življenja ljudi po svetu. Nukleotidno zaporedje virusa SARS-CoV-2 je 79.6% enako virusu SARS CoV, ki je povzročil epidemijo SARS. Domena SUD M proteina Nsp3 SARS-CoV z aminokislinskimi ostanki K563, K565, K568 in E571 veže G-kvadruplekse. Ker mutacija vezavnega mesta prepreči replikacijo virusa SARS-CoV in vivo, ima interakcija z G-kvadrupleksom velik terapevtski potencial. Primerjali smo aminokislinsko zaporedje domene SUD M SARS-CoV in SARS-CoV-2 in ugotovili, da je vezavno mesto K563, K565, K568 in E571 ohranjeno pri virusu SARS-CoV-2. Ker SARS-CoV-2 ne vsebuje genomskih zaporedij, ki bi tvorila stabilne G-kvadruplekse, je verjetno, da so tarče SUD M celične z gvanini bogate mRNA. Analizirali smo 3' neprevedene regije genov MAPK1, RAB6B, BBC3 in TAB3 ter z enodimenzionalnimi NMR eksperimenti dokazali, da 21 nukleotidov dolgo z gvanini bogato zaporedje RAB6B tvori G-kvadrupleksne strukture. S podaljšanjem zaporedja z uracilnimi nukleotidi smo zmanjšali agregacijo oligonukleotida. Z dvodimenzionalnimi NMR metodami smo pokazali, da 19 aminokislin dolg peptid, ki ustreza zaporedju vezavnega mesta za G-kvadrupleks domene SUD M SARS-COV-2 interagira z G-kvadrupleksno RAB6B RNA.

• Language: Slovenian
• Keywords: SUD M, SARS-CoV-2, G-kvadrupleks, NMR
• Work type: Master's thesis/paper
• Typology: 2.09 - Master's Thesis
• Organization: FKKT - Faculty of Chemistry and Chemical Technology
• Year: 2023
• PID: 20.500.12556/RUL-144875
• COBISS.SI-ID: 146292483
• Publication date in RUL: 20.03.2023

Secondary language

• Language: English
• Title: Human 3’-UTR mRNA interactions with peptides of SUD M domain of the SARS-CoV-2 Nsp3 protein

Abstract

SARS-CoV-2 pandemic has caused major negative socio-economic consequences around the globe. SARS-CoV-2 shares a 79.6% nucleotide sequence identity with the SARS-CoV virus that was the cause of the SARS epidemic. Amino acid residues K563, K565, K568 and E571 of the SUD M domain of Nsp3 SARS-CoV bind G-quadruplexes. Because mutations in this binding sequence abolish SARS-CoV viral replication in vivo, the interaction of SUD M with G-quadruplexes possesses great therapeutic potential. A comparison of the SUD M domains of both Coronaviruses revealed an identical G-quadruplex binding sequence in the SARS-CoV-2 SUD M. However, no stable G-quadruplex structures were found in SARS-CoV-2 genomic sequences, and it is therefore believed that cellular G-quadruplex mRNAs are the target of SUD M. 3'UTR mRNA sequences of the genes MAPK1, RAB6B, BBC3 in TAB3 were analysed and a 21-nucleotide guanine-rich sequence from the RAB6B gene was found to form G-quadruplexes using one-dimensional solution NMR. Aggregation was minimised by the addition of uracil residues at the 5’- and 3’-end. Using two-dimensional NMR experiments we have demonstrated that a 19 amino acid peptide of the SARS-CoV-2 SUD M and G-quadruplex RAB6B RNA interact in vitro.

• Keywords: SUD M, SARS-CoV-2, G-quadruplex, NMR