Unutarstanične interakcije adenovirusnog E3-14.7K proteina s proteinima NF-kappaB signalnog puta

master's thesis

• Title (english): Intracellular interactions of Adenoviral E3-14.7K protein with NF-kappaB proteins
• Author: Vedrana Bukal
• Mentor: Andreja Ambriović Ristov (mentor)
• Granter: University of Zagreb; Faculty of Food Technology and Biotechnology; Zagreb
• Defense date and country: 2012-07-02, Croatia
• Scientific / art field, discipline and subdiscipline: BIOTECHNICAL SCIENCES; Biotechnology

Abstract

Adenovirusi sadrže mnogo gena koji utječu na imunološki sustav domaćina i suzbijaju njegov odgovor, te se mnogi od tih gena nalaze u ranoj regiji 3 (E3). U ovom radu smo se orijentirali na E3-14.7K imunoregulatorni protein i njegov utjecaj na NF-κB signalni put. NF-κB transkripcijski faktori su glavni regulatori gena zaduženih za imunosni i upalni odgovor, te gena zaduženih za preživljavanje. Ovdje smo pokušali rasvijetliti utjecaj Ad E3-14.7K proteina na NF-κB promotor, utvrditi njegove interakcije s p50 i p65 podjedinicom ovog signalnog puta, kao i s peptidima izvedenim iz E3-14.7K proteina. Nakon transfekcije stanica s E3-14.7K proteinom, zabilježena je aktivacija NF-κB promotora, te je aktivacija bila značajnija nakon tretmana s TNF-α koji je jedan od aktivatora NF-κB signalnog puta. Interakcije između E3-14.7K i p50, te p65 podjedinice su detektirane pomoću BRET tehnike, te je pomoću konfokalnog mikroskopa utvrđeno da E3-14.7K ima tendenciju stupanja u interakcije s p65 podjedinicom.

Abstract (english)

Adenoviruses have a variety of immunoregulatory genes, and many of them are located in early region 3 (E3). In our research we were oriented on E3-14.7K immunoregulatory protein and its influence on NF-κB pathway. NF-κB family of transcription factors are main regulators of immune, inflammatory and cell survival associated genes. We were trying to elucidate influence of Ad E3-14.7K protein on NF-κB promoter, detect its interactions with p50 and p65, as well as for peptides derived from E3-14.7K. Increase in activation of NF-κB promoter was noticed after transfection of cells with E3-14.7K, and this increase was even more significant after the treatment with human TNF – α which is one of the activators of NF-κB pathway. With BRET assay we were able to detect interactions between E3-14.7K and p50, and between E3-14.7K and p65 subunit of NF-κB pathway. E3-14.7K has shown preference to interact with p65, as we confirmed with confocal microscopy.

• Keywords: NF-κB signalni put; Ad E3-14.7K; BRET tehnika; p50; p65.
• Keywords (english): NF-κB pathway; E3-14.7K; BRET assay; p50; p65.
• Language: croatian
• URN:NBN: urn:nbn:hr:159:476606
• Study programme: Title: Molecular Biotechnology; Study programme type: university; Study level: graduate; Academic / professional title: magistar/magistra inženjer/inženjerka molekularne biotehnologije (magistar/magistra inženjer/inženjerka molekularne biotehnologije)
• Type of resource: Text
• File origin: Born digital
• Access conditions: Access restricted to students and staff of home institution
• Created on: 2017-05-08 12:53:13