magistrsko delo
Sara Laznik (Author), Janez Plavec (Mentor), Miha Pavšič (Thesis defence commission member), Jurij Lah (Thesis defence commission member)

Abstract

G-kvadrupleks se lahko potencialno tvori na dolgi nekodirajoči RNA psevdogena REG1CP, ki interagira s helikazo FANCJ. Interakcija vpliva na transkripcijo gena REG3A, kar je povezano z napredovanjem raka debelega črevesa. V magistrskem delu smo dokazali, da izbrano zaporedje iz dolge nekodirajoče RNA REG1CP tvori G kvadrupleks s tremi naloženimi G kvarteti. Pri sobni temperaturi je oligonukleotid v raztopini obstajal v dveh sekundarnih oblikah, ki sta v ravnotežju, in sicer kot lasnica in G kvadrupleks RNA. Pri fiziološki temperaturi se je to ravnotežje pomaknilo proti tvorbi G kvadrupleksa, kar nakazuje na to, da je tudi in vivo bolj zastopan G kvadrupleks. Iz zaporedja dolge nekodirajoče RNA (lncRNA) smo sintetizirali različno dolge oligonukleotide RNA in z zamenjavami posameznih nukleotidov dobili zaporedje, ki ima dobro ločene resonance v NMR spektru. Določili smo njegovo termično stabilnost in s pomočjo NMR spektroskopije proučevali 3D strukturo G-kvadrupleksa REG1CP. S pomočjo CD spektroskopije smo ugotovili, da ima omenjeni G-kvadrupleks paralelno topologijo. Prav tako smo G-kvadrupleks titrirali z različnimi peptidi in spremljali interakcije G kvadrupleks:peptid. S peptidoma Rhau18 in Rhau23 je G-kvadrupleks REG1CP tvoril kompleks. S peptidom iz zaporedja helikaze FANCJ je z G kvadrupleksom prišlo do šibkejše interakcije.

Keywords

G-kvadrupleksi;lncRNA;NMR spektroskopija;proteini;peptidi;FANCJ;tumorigenost;magistrska dela;

Data

Language: Slovenian
Year of publishing:
Typology: 2.09 - Master's Thesis
Organization: UL FKKT - Faculty of Chemistry and Chemical Technology
Publisher: [S. Laznik]
UDC: 577.21(043.2)
COBISS: 159777283 Link will open in a new window
Views: 46
Downloads: 17
Average score: 0 (0 votes)
Metadata: JSON JSON-RDF JSON-LD TURTLE N-TRIPLES XML RDFA MICRODATA DC-XML DC-RDF RDF

Other data

Secondary language: English
Secondary title: Structural study of G-quadruplex formed by long non-coding RNA of pseudogene REG1CP
Secondary abstract: G-quadruplex can potentially form on the long non-coding RNA of pseudogene REG1CP, which interacts with the FANCJ helicase. This interaction affects the transcription of the REG3A gene, which is associated with the progression of colon cancer. In the master's thesis, we proved that the selected sequence from the long non coding RNA REG1CP forms a G-quadruplex with three stacked G quartets. At room temperature, the oligonucleotide existed in two secondary forms, as a hairpin and a G quadruplex, which were in equilibrium. At the physiological temperature, this equilibrium shifted towards the formation of the G-quadruplex, indicating that the G quadruplex is more prevalent in vivo. We synthesized RNA oligonucleotides of different lengths from the lncRNA sequence. By substituting individual nucleotides, we obtained a sequence with well-separated resonances in the NMR spectrum. We determined its thermal stability and studied the 3D structure of the G quadruplex REG1CP with the help of different NMR spectroscopy experiments. With the CD spectroscopy, we found that the mentioned G quadruplex has a parallel topology. Additionally, we titrated the G-quadruplex with different peptides and monitored their interactions. The G-quadruplex REG1CP formed a complex with Rhau18 and Rhau23 peptides, while the interaction with the peptide from the FANCJ helicase was weaker.
Secondary keywords: G-quadruplex;incRNA;NMR spectroscopy;FANCJ;tumorigenicity;RNK;Univerzitetna in visokošolska dela;
Type (COBISS): Master's thesis/paper
Study programme: 1000377
Embargo end date (OpenAIRE): 1970-01-01
Thesis comment: Univ. v Ljubljani, Fak. za kemijo in kemijsko tehnologijo, smer Biokemija
Pages: 53 str.
ID: 19336056