• Title/Summary/Keyword: Glu-AdT

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Growth Inhibition of Escherichia coli during Heterologous Expression of Bacillus subtilis Glutamyl-tRNA Synthetase that Catalyzes the Formation of Mischarged Glutamyl-$tRNA_{l}$$^{Gln}$

  • Baick, Ji-Won;Yoon, Jang-Ho;Suk Namgoong;Dieter Soll;Kim, Sung-Il;Eom, Soo-Hyun;Hong, Kwang-Won
    • Journal of Microbiology
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    • v.42 no.2
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    • pp.111-116
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    • 2004
  • It is known that Bacillus subtilis glutamyl-tRNA synthetase (GluRS) mischarges E. coli $tRNA_{1}$$^{Gln}$ with glutamate in vitro. It has also been established that the expression of B. subtilis GluRS in Escherichia coli results in the death of the host cell. To ascertain whether E. coli growth inhibition caused by B. subtilis GluRS synthesis is a consequence of Glu-$tRNA_{1}$$^{Gln}$ formation, we constructed an in vivo test system, in which B. subtilis GluRS gene expression is controlled by IPTG. Such a system permits the investigation of factors affecting E. coli growth. Expression of E. coli glutaminyl-tRNA synthetase (GlnRS) also amelio-rated growth inhibition, presumably by competitively preventing $tRNA_{1}$$^{Gln}$ misacylation. However, when amounts of up to 10 mM L-glutamine, the cognate amino acid for acylation of $tRNA_{1}$$^{Gln}$, were added to the growth medium, cell growth was unaffected. Overexpression of the B. subtilis gatCAB gene encoding Glu-$tRNA^{Gln}$ amidotransferase (Glu-AdT) rescued cells from toxic effects caused by the formation of the mis-charging GluRS. This result indicates that B. subtilis Glu-AdT recognizes the mischarged E. coli Glu-$tRNA_{1}$$^{Gln}$, and converts it to the cognate Gln-$tRNA_{1}$$^{Gln}$ species. B. subtilis GluRS-dependent Glu-$tRNA_{1}$$^{Gln}$ formation may cause growth inhibition in the transformed E. coli strain, possibly due to abnormal protein synthesis.

Cloning of the $Glu-tRNA^{Gln}$ Amidotransferase (gatCAB) Gene from Staphylococcus aureus

  • Namgoong, Suk;Hong, Kwang-Won;Lee, Se-Yong
    • Journal of Microbiology and Biotechnology
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    • v.11 no.2
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    • pp.251-258
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    • 2001
  • In Gram(+) bacteria and organelles in higher eukarotes, $Gln-tRNA^{Gln}$ utilized for protein biosynthesis is formed by a tRNA-dependent amino acid transformation using mischarged $Gln-tRNA^{Gln}$ as the intermediate. In this study, the gatCAB gene encoding $Gln-tRNA^{Gln}$ amidotransferase (Glu-AdT) of Staphylococcus aureus was cloned and its nucleotide sequence wa determined. The S. aureus gatCAB gene was organized in an operon structure consisting of three open reading frames (gatC, gatA, and gatB), similar to that of Bacillus subtilis. The gene sequences for the A and B subunits of$Gln-tRNA^{Gln}$ amidotransferase showed significant homology (77 and 87% homology with amino acid sequence) with the gatA and gatB genes of B. subtilis, yet the C subunit (gatC) showed a relatively lowe homology with the B. subtilis gatC gene and other orthologues. The cloned S. aureus <$Gln-tRNA^{Gln}$ amidotransferase gene was highly expressed in Escherichia coli, and the resulting crude enzyme could convert misacylated <$Gln-tRNA^{Gln}$ into $Gln-tRNA^{Gln}$ in vitro.

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Effects of Selective Thromboxane $A_2$-Receptor Antagonist, KT2-962 on Adriamycin-induced Nephrotoxicity in Rats (흰쥐에서 Adriamycin-유발 신독성에 대한 Thromboxane $A_2$ 수용체 길항제인 KT2-962의 효과)

  • 문삼영;이순복;신현진;고현철;엄애선;강주섭
    • Biomolecules & Therapeutics
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    • v.8 no.3
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    • pp.241-247
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    • 2000
  • The present study was designed to assess the protective effect of a selective thromboxane $A_2$ receptor antagonist, KT2-962 (KT2) and possible mechanisms of adriamycin(AD)-induced nephrotoxicity in rats. The male Wistar rats were given either of AD (7.5 mg/kg, i.v.) alone in the AD-group (n=5) or in KT2+AD- group (n=5) which is a combination of AD and KT2 (30 mg/kg/day, i.p.) for 10 days from 3 days before and 7 days after AD injection. The body weight, 24-hours urine volume, urine protein and urinary N-acetyl-$\beta$-D-glu-cosaminidase (NAG) activity were measured with an interval of 2 days during 1 week. BUN, serum creatinine and creatinine clearance were measured on the 7th day. KT2 has significantly suppressed AD-induced change of body weight, 24-hours urine volume, urine protein and urinary NAG activity in the KT2+AD-group. The change of BUN, serum creatinine and creatinine clearance were significantly inhibited in the B7T2+AD-group. Based on these results, it is concluded that KT2 prevents AD-induced nephrotoxicity and suggests that endogenous thromboxane A2 may play an important role in AD-induced nephrotoxicity in rats.

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The Effect of Feeding with Sasa quelpaertensis Nakai Extract on Change in Economic Traits of the Pig (제주조릿대 (Sasa quelpaertensis Nakai) 추출물 급여가 돼지의 경제형질 특성 변화에 미치는 영향)

  • Hyeon Ah Kim;Sang Hwi Im;Ju Sung Kim;Mi Hyeon Park;Jong An Lee;Yong Jun Kang;In Cheol Cho;Moon Cheol Shin
    • Journal of The Korean Society of Grassland and Forage Science
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    • v.43 no.4
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    • pp.240-247
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    • 2023
  • This experiment investigated the effects of feed additives of Sasa quelpaertensis Nakai (SQN) extract on Landrace pigs on economic traits such as the quality, physiological characteristics, and productivity. Sixteen pigs with an average age of 154 days were selected as experimental subjects. The experiment was conducted by dividing the group into eight pigs for the supplementation group, feeding with SQN extract, and another eight for the control group feeding without SQN extract. Water was fed ad libitum. On the 30th day, there was no significant difference between meat quality and productivity. However, the glucose and thyroxine were statistically lower with the supplementation group than with the control group (p<0.05). Also, the levels of creatinine difference between 1.18 ± 0.12 mg/㎗ with the supplementation group and 0.70 ± 0.06 mg/㎗ with the control group (p<0.05). However, all serum biochemistry values were within a normal range, with no health problems. The present study will help solve the problem of reducing the diversity of plant species in Halla Mountain by increasing the availability of the SQN as a pig feed additive.