Razvoj in vivo sustava za testiranje inhibicije triptofanil-tRNA sintetaze

master's thesis

• Title (english): Development of an in vivo tryptofanyl-tRNA synthetase inhibition assay
• Author: Kristian Subota
• Mentor: Marko Močibob (mentor)
• Committee member: Marko Močibob (predsjednik povjerenstva)
• Committee member: Petra Korać (član povjerenstva)
• Committee member: Ivana Ivančić Baće (član povjerenstva)
• Committee member: Inga Urlić (član povjerenstva)
• Granter: University of Zagreb; Faculty of Science; (Department of Biology); Zagreb
• Defense date and country: 2025-02-07, Croatia
• Scientific / art field, discipline and subdiscipline: NATURAL SCIENCES; Biology

Abstract

Aminoacil-tRNA-sintetaze (AaRS), predstavljaju dobru metu za razvoj novih antibiotika, jer
postoje značajne razlike između evolucijski očuvanih prokariotskih i eukariotskih enzima. Većina
bakterija koristi jednu konstitutivno eksprimiranu triptofanil-tRNA-sintetazu (TrpRS), no Bacillus
thuringiensis ima i bona fide inducibilnu TrpRS2, kojoj je zasad uloga nepoznata. U prijašnjim
istraživanjima na izoleucil-tRNA-sintetazi je pokazano da je IleRS2 inducibilna te daje bakterijama
otpornost na... More mupirocin, dok konstitutivna verzija ima bolju učinkovitost u aminoacilaciji.
Postavljena je hipoteza da bi TrpRS2 mogla imati sličnu ulogu. Stoga je klonirana promotorska
regija gena za TrpRS2 iz B. thuringiensis u Bacillus subtilis s povezanim reporterskim genom
(lacZ). Ovaj sintetski sustav omogućio bi brzi probir prirodnih spojeva koji mogu inhibirati
triptofanil-tRNA-sintetazu, što bi moglo doprinijeti razvoju novih antibiotika. Do sada je u
kliničkoj primjeni samo tri inhibitora AaRS: mupirocin, halofuginon i tavaborol. Less

Abstract (english)

Aminoacyl-tRNA-synthetases (AaRS) represent a good target for the development of new
antibiotics, because there are significant differences between evolutionarily conserved prokaryotic
and eukaryotic enzymes. Most bacteria use one constitutively expressed tryptophanyl-tRNAsynthetase (TrpRS), but Bacillus thuringiensis also has a bona fide inducible TrpRS2, whose role
is currently unknown. In previous research, on isoleucyl-tRNA-synthetase, it was shown that
IleRS2 is responsible for... More bacterial resistance to mupirocin, while the constitutive version has better
efficiency in aminoacylation. It was hypothesized that TrpRS2 might play a similar role. Therefore,
the promoter region of the TrpRS2 gene from B. thuringiensis was cloned into Bacillus subtilis
with an associated reporter gene (lacZ). This synthetic system would enable rapid screening of
natural compounds that can inhibit tryptophanyl-tRNA-synthetase, which could contribute to the
development of new antibiotics. So far, only three AaRS inhibitors are in clinical use: mupirocin,
halofuginone and tavaborole. Less

• Keywords: aminoacil-tRNA-sintetaze; antibiotici; genetičko inženjerstvo; triptofan
• Keywords (english): aminoacyl-tRNA-synthetases; antibiotics; genetic engineering; tryptophan
• Language: croatian
• URN:NBN: urn:nbn:hr:217:154981
• Project: Number: IP-2022-10-1400; Title: Istraživanja aminoacil-tRNA-sintetaza kao antibiotskih meta i mehanizama kojima razvijaju rezistenciju; Title: Investigation of aminoacyl-tRNA syntetases as targets for antibiotics and how they develop resistance mechanisms; Acronym: ARSTARE; Leader: Ita Gruić-Sovulj; Jurisdiction: Croatia; Funder: Hrvatska zaklada za znanost; Funding stream: Research Projects
• Study programme: Title: Molecular Biology-University graduate programme; Study programme type: university; Study level: graduate; Academic / professional title: sveučilišni/a magistar/magistra molekularne biologije (sveučilišni/a magistar/magistra molekularne biologije)
• Type of resource: Text
• File origin: Born digital
• Access conditions: Open access
• Created on: 2025-02-27 12:22:09