Proteinske domene aminoacil-tRNAsintetaza i interakcije s drugim proteinima

master's thesis

• Title (english): Protein domains of aminoacyl-tRNA synthetases and interactions with other proteins
• Author: Marija Podolski
• Mentor: Ivana Weygand-Đurašević (mentor)
• Committee member: Ivana Weygand-Đurašević (predsjednik povjerenstva)
• Committee member: Mirjana Kalafatić (član povjerenstva)
• Committee member: Dijana Škorić (član povjerenstva)
• Granter: University of Zagreb; Faculty of Science; (Department of Biology); Zagreb
• Defense date and country: 2009, Croatia
• Scientific / art field, discipline and subdiscipline: NATURAL SCIENCES; Biology

Abstract

Prethodno otkrivena kristalna struktura seril-tRNA-sintetaze (SerRS) metanogenog tipa iz arheje Methanosarcina barkeri ukazala je na neke atipične karakteristike ovog enzima, jedna od kojih je bila i prisutnost motiva HTH unutar C-terminalne domene proteina. Za njega se pretpostavljalo da je ima važnu ulogu pri dimerizaciji SerRS i interakciji između N- i Cterminalnih domena proteina. Ova pretpostavka je potvrđena našim rezultatima dobivenim metodom dvaju hibrida. Osim toga, pretraživanje metodom dvaju hibrida iskorišteno je za utvrđivanje proteina s kojima SerRS iz Methanothermobacter thremautotrophicus interagira, jedan od kojih je bila i arginiltRNA- sintetaza (ArgRS). Ovu interakciju smo potvrdili afinitetnom kromatografijom („GSTpulldown assay“). Prethodno je pronađeno da neke SerRS metanogenog tipa mogu, osim pripadnih tRNASer i tRNASec prepoznati i eukariotske i prokariotske tRNASer. Naši rezultati predlažu da je to slučaj i kod SerRS iz M. thermautotrophicus.

Abstract (english)

Recently resolved crystal structure of methanogenic type seryl-tRNA synthetase (SerRS) from Methanosarcina barkeri revealed some atypical features, one of which was an idiosyncratic helix-turn-helix motif within C-terminal domain of the protein. This motif seems to be important for dimerization of the enzyme and interaction between Nand C-terminal domains. This assumption was supported by our experimental results achieved by yeast two-hybrid assays. In addition, the yeast two-hybrid approach was used to screen for proteins interacting with SerRS in Methanothermobacter thermautotrophicus. One of the proteins which was found to interact with SerRS was arginyl-tRNA synthetase. To support this finding, we proved this interaction by affinity chromatography (GST-pulldown assay). As it was established for some methanogenic type SerRSs previously, some of these enzymes are capable of recognizing both eukaryotic and prokariotyc tRNASer in addition to the homologous tRNASer and tRNASec substrates. Our results indicate that this feature is also displayed by M.thermautotrophicus SerRS.

• Keywords: seril-tRNA-sintetaza; proteinske interakcije; metanogene arheje; sustav dvaju hibrida
• Keywords (english): seryl-tRNA synthetase; protein-protein interactions; methanogenic archaea; yeast two-hybrid system
• Language: croatian
• URN:NBN: urn:nbn:hr:217:521645
• Study programme: Title: Molecular Biology; Study programme type: university; Study level: graduate; Academic / professional title: magistar/magistra molekularne biologije (magistar/magistra molekularne biologije)
• Type of resource: Text
• File origin: Born digital
• Access conditions: Open access
• Created on: 2018-03-15 09:43:19