Eukaryotic features of isoleucyl-tRNA synthetase from bacteria Streptomyces griseus

Biluš, Mirna (2013) Eukaryotic features of isoleucyl-tRNA synthetase from bacteria Streptomyces griseus. Diploma thesis, Faculty of Science > Department of Biology.

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Abstract

Isoleucyl-tRNA synthetases (IleRS) catalyze the formation of Ile-tRNAIle. Eukaryotic IleRSs differ from prokaryotic homologues by primary structures, resistance to antibiotic mupirocin and substrate specificity. Mupirocin strongly inhibits IleRS from numerous bacteria, however it does not affect eukaryotes. However, IleRS from some prokaryotes, including Streptomyces griseus, align with the eukaryotic enzymes. The gene for IleRS from S. griseus was cloned and expressed in Escherichia coli. The protein was purified by affinity chromatography. Kinetic characterization established the ability of S. griseus IleRS to heterologously aminoacylate yeast and E. coli tRNAs. As prokaryotic IleRS (E. coli IleRS) does not aminoacylate yeast tRNAIle, the results indicate that S. griseus IleRS resembles the yeast homologue which aminoacylates E. coli tRNA. Diffusion tests showed that like the eukaryotic homologue from yeast, S. griseus IleRS is resistant to mupirocin. Additionally, S. griseus IleRS was able to complement the growth of E. coli in the presence of mupirocin.

Item Type: Thesis (Diploma thesis)
Keywords: aminoacyl-tRNA synthetases, complementation, identity elements, mupirocin, tRNA
Supervisor: Gruić Sovulj, Ita
Date: 2013
Number of Pages: 68
Subjects: NATURAL SCIENCES > Biology
Divisions: Faculty of Science > Department of Biology
Depositing User: Silvana Šehić
Date Deposited: 29 Sep 2014 12:21
Last Modified: 29 Sep 2014 12:21
URI: http://digre.pmf.unizg.hr/id/eprint/2861

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