• Journal of Microbiology and Biotechnology
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• v.20 no.9
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• pp.1283-1287
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• 2010

In eukaryotes, eRF3 participates in translation termination and belongs to the superfamily of GTPases. In this work, the dissociation constants for nucleosides bound to Euplotes octocarinatus eRF3 in the presence and absence of eRF1a were determined using fluorescence spectra methods. Furthermore, a GTP hydrolyzing assay of eRF3 was carried out using an HPLC method, and the kinetic parameters for GTP hydrolysis by eRF3 were determined. Consistent with data from humans, the results showed that eRF1a promoted the binding of GTP to eRF3 and the GTP hydrolyzing activity of eRF3. However, in contrast to the lack of GTP binding in the absence of eRF1 in human eRF3, the E. octocarinatus eRF3 was able to bind GTP by itself. The nucleotide binding affinity of the E. octocarinatus eRF3 also differed from the human data. A structure model and amino acid sequence alignment of potential G domains indicated that these differences may be due to valine 317 and glutamate 452 displacing the conserved glycine and lysine involved in GTP binding.

• Journal of Species Research
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• v.12 no.3
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• pp.203-211
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• 2023

Five ciliate species of Euplotes were isolated from fresh and coastal water during a sampling survey to identify unrecorded ciliates in South Korea. Their morphology was investigated using live observation, protargol and "wet" silver nitrate staining methods. Brief descriptions and microphotographs of each species and a comparison with related species are provided. Euplotes focardii is characterized by an average size of 65×47 ㎛ after protargol impregnation, 6 dorsal and 3 ventral ridges and dorsal argyrome pattern of double-eurystomus type. Euplotes nobilii shows an average size of 34×20 ㎛ after protargol staining, 6 dorsal and 3 ventral ridges and dorsal argyrome pattern of double-patella type. Euplotes octocarinatus, the only freshwater species described in the present study, is characterized by an average size of 66×46 ㎛ after protargol impregnation, 6 dorsal and 3 ventral ridges and dorsal argyrome pattern of double-patella type. Euplotes petzi has an average size of 43×30 ㎛ after protargol staining, a macronucleus hook-shaped and dorsal argyrome pattern in double-patella type. Euplotes raikovi is characterized by an average size of 40×24 ㎛ after protargol staining, 6 dorsal and 3 ventral ridges and dorsal argyrome pattern of double-patella type.

• Journal of Microbiology and Biotechnology
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• v.22 no.2
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• pp.176-183
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• 2012

Eukaryotic translation termination is governed by eRF1 and eRF3. eRF1 recognizes the stop codons and then hydrolyzes peptidyl-tRNA. eRF3, which facilitates the termination process, belongs to the GTPase superfamily. In this study, the effect of the MC domain of eRF1a (eRF1aMC) on the GTPase activity of eRF3 was analyzed using fluorescence spectra and high-performance liquid chromatography. The results indicated eRF1aMC promotes the GTPase activity of eRF3, which is similar to the role of eRF1a. Furthermore, the increased affinity of eRF3 for GTP induced by eRF1aMC was dependent on the concentration of $Mg^{2+}$. Changes in the secondary structure of eRF3C after binding GTP/GDP were detected by CD spectroscopy. The results revealed changes of conformation during formation of the eRF3C GTP complex that were detected in the presence of eRF1a or eRF1aMC. The conformations of the eRF3C eRF1a GTP and eRF3C eRF1aMC GTP complexes were further altered upon the addition of $Mg^{2+}$. By contrast, there was no change in the conformation of GTP bound to free eRF3C or the eRF3C eRF1aN complex. These results suggest that alterations in the conformation of GTP bound to eRF3 is dependent on eRF1a and $Mg^{2+}$, whereas the MC domain of eRF1a is responsible for the change in the conformation of GTP bound to eRF3 in Euplotes octocarinatus.