Methionine- R-reductase plays a defensive role against cadmium toxicity in Schizosaccharomyces pombe

Hannah Jo,Kisup Ahn,Chang-Jin Lim 강원대학교 기초과학연구소 2012 기초과학연구 Vol.23 No.-

Methionine sulfoxide reductases (Msrs) are oxidoreductases which catalyze thiol-dependent reduction of free and protein- bound methionine sulfoxides back to corresponding methionine. Methionine-S-sulfoxide (Met-S-SO) and methionine-R-sulfoxide (Met-R-SO) are reduced by methionine-S-sulfoxiede reductase (MsrA) and methionine-R-sulfoxide reductase (MsrB), respectively. In the present study, the cellular functions of MsrB were further characterized using the MsrB-overexpressing recombinant plasmid pFMetSO in the fission yeast. The S. pombe cells harboring pFMetSO exhibited better growth than the vector control cells, when shifted into the fresh medium containing cadmium. In the presence of different levels of cadmium, the S. pombe cells harboring pFMetSO contained diminished NO and reactive oxygen species (RS) levels than the vector control cells. On the contrary, the S. pombe cells harboring pFMetSO contained higher levels of total glutathione (GSH) than the vector control cells under the same treatments. Taken together, the S. pombe MsrB plays a protective role against cadmium toxicity.

Protective Role of Schizosaccharomyces pombe Spp1 against Oxidative Stress Induced by Peroxide Compounds

Min-Sik Park,Kisup Ahn,Chang-Jin Lim 강원대학교 기초과학연구소 2012 기초과학연구 Vol.23 No.-

The Schizosaccharomyces pombe spp1⁺ gene encoding a putative member of the PPPDE superfamily was cloned into an E. coli-yeast shuttle vector pRS316, which resulted in the recombinant plasmid pYSTP. The encoded sequence shares a homology with the counterparts of Arabidopsis thaliana, Picea sitchensis and Chaetomium globosum. The S. pombe cells harboring pYSYP were shown to contain a significantly higher level of the spp1⁺ mRNA than the control cells, indicating that the cloned gene is genuinely functional, and they showed a better growth n liquid media than the control cells. The S. pombe cells harboring pYSTP gave rise to an enhanced survival in liquid medium with hydrogen peroxide (H₂O₂), tert-buty1 bydroeroxide (t-BOOH) or tert-butylhydroquinone (t-BHQ) than the vector control cells. Under stress, the S. pombe cells harboring pYSTP showed slighly lower reactive oxygen species(ROS) level than the vector control cells. The total glutathione (GSH) content is higher in the S. pombe cells harboring pYSTP than the vector control cells. In brief, the S. pombe spp1⁺ gene plays a protective role in the defensive response to oxidative stress.

Polymorphism in Intrinsic Antibiotic Resistance of Azospirillum Isolates from Ornamental Plants

Gadagi, Ravi,Krishnaraj, P.U.,Kulkarni, J.H.,Ahn, Kisup,Sa, Tongmin 한국환경농학회 2001 한국환경농학회지 Vol.20 No.5

The polymorphism of Azospirillum isolates from ornamental rhizosphere and two reference strains were examined with respect to intrinsic antibiotic resistance (IAR) profile. All the isolates showed different intrinsic resistances to different antibiotics viz., tetracycline, kanamycin, nalidixic acid, streptomycin, ampicillin, spectinomycin and chloramphenicol. All the strains demonstrated susceptibility to high concentration of all antibiotics used in the present experiment. In addition to these general patterns, we also obseved the multiple antibiotic resistances of Azospirillum strains. The Azospirillum sp. OAD-11 was resistant to tetracycline, streptomycin and ampicillin, and Azospirillum sp. OAD-57 was resistant to tetracycline and streptomycin. Conversely, Azospirillum sp. OAD-9 possessed the dual susceptibility to tetracycline and spectinomycin, whereas Azospirillum sp. OAD-37 was dual susceptible to streptomycin and kanamycin. Such multiple antibiotic resistant/susceptible traits could be useful for the identification of the strains in field experiments or in molecular genetic transfer experiments.

Differential Regulation of Three Genes Encoding Glutathione S-Transferases in Schizosaccharomyces pombe

Chang-Jin Lim,Hong-Gyum Kim,김병철,박은희,Kisup Ahn 한국분자세포생물학회 2004 Molecules and cells Vol.18 No.3

Glutathione S-transferases (GSTs) are detoxifying enzymes that catalyze the conjugation of glutathione with a variety of reactive electrophilic compounds. Three GST genes were previously characterized in the fission yeast Schizosaccharomyces pombe. In this work, we examined the transcriptional regulation of these genes using individual GST-lacZ fusions and RT-PCR. Basal synthesis of β-galactosidase from the GSTII-lacZ fusion was higher than from the GSTI-lacZ and GSTIII-lacZ fusion. Diethylmaleate (0.2 mM) greatly enhanced the synthesis of β-galactosidase from the GSTII-lacZ fusion, but did not affect synthesis from the other two fusion genes. A switch to 0.3% glucose or 0.3% sucrose as sole carbon source enhanced expression from the GSTIIIlacZ fusion gene, while sodium nitroprusside (1.5 mM), tert-butylhydroquinone (0.2 mM), and L-buthionine- [S,R]-sulfoximine (0.01 mM) increased expression of the GSTII gene. The effects of these agents on GST mRNA levels were confirmed by measurements employing RTPCR. Our results suggest that transcription of the three S. pombe GST genes is subjected to differential regulation under various stress conditions, and may be linked to their different physiological functions.

Pap1p-dependent upregulation of thioredoxin 3 and thioredoxin reductase genes from the fission yeast under nitrosative stress.

Park, Min-Sik,Kim, Hyeon-Jung,Park, A Rum,Ahn, Kisup,Lim, Hye-Won,Lim, Chang-Jin National Research Council 2012 Canadian journal of microbiology Vol.58 No.2

<P>The thioredoxin system, consisting of thioredoxin, thioredoxin reductase, and NADPH, is involved in the response against a variety of stresses. The TRX3(+) and TrxR(+) genes encode thioredoxin 3 and thioredoxin reductase, respectively, in the fission yeast Schizosaccharomyces pombe . Their transcriptional regulations were studied using the lacZ fusion genes. Synthesis of β-galactosidase from the TRX3(+)-lacZ fusion gene was markedly enhanced by nitric-oxide-generating sodium nitroprusside in the Pap1p-positive cells but not in the Pap1p-negative cells. Similarly, synthesis of β-galactosidase from the TrxR(+)-lacZ fusion gene was upregulated by sodium nitroprusside in a Pap1p-dependent manner. Synthesis of β-galactosidase from the TRX3(+)-lacZ and TrxR(+)-lacZ fusion genes was also enhanced by S-nitrosoglutathione in the Pap1p-positive cells but not in the Pap1p-negative cells. In brief, the S. pombe genes encoding thioredoxin 3 and thioredoxin reductase are upregulated under nitrosative stress in a Pap1p-dependent manner.</P>

Performance of MPS Bacterial Inoculation in Two Consecutive Growth of Maize Plants

Park, Myungsu,Gadagi, Ravi,Singvilay, Olayvanh,Kim, Chungwoo,Chung, Heekyung,Ahn, Kisup,Sa, Tongmin 한국환경농학회 2001 한국환경농학회지 Vol.20 No.5

Two successive in vitro experiments were carried out to examine the effect of MPS bacterial inoculation on growth, and nitrogen and phosphorus accumulation of maize plants under greenhouse condition in the same soil. There were four treatments, uninoculated control and three phosphate solubilizing bacterial inoculations, viz., Pseudomonas striata, Burkholderia cepacia and Serratia marcescens. The inoculated plants showed the higher plant height, total dry mass, nitrogen and phosphorus accumulation when compared to uninoculated control plants in both experiments. In the combined data analysis from two experiments, the plants inoculated with P. striata and B. cepacia showed significantly higher plant height, total dry mass and P accumulation when compared to S. marcescens inoculated plant and uninoculated control plants. The P. striata and B. cepacia inoculation enhanced total dry matter accumulation by 14% and phosphorus accumulation by 25% over the uninoculated control plants. The nitrogen and phosphorus concentration of maize plants were also increased due to MPS bacterial inoculation, however, the effect was not significant.

Effects of Heterologous Expression of Thioredoxin Reductase on the Level of Reactive Oxygen Species in COS-7 Cells

Chang-Jin Lim,Hyun-Jung Kang,Sung-Min Hong,김병철,박은희,Kisup Ahn 한국분자세포생물학회 2006 Molecules and cells Vol.22 No.1

Anti-inflammatory activity of n-propyl gallate through down-regulation of NF-관B and JNK pathways.

Jung, Hyun-Joo,Kim, Su-Jung,Jeon, Woo-Kwang,Kim, Byung-Chul,Ahn, Kisup,Kim, Kyunghoon,Kim, Young-Myeong,Park, Eun-Hee,Lim, Chang-Jin Kluwer Academic/Plenum Publishers 2011 INFLAMMATION Vol.34 No.5

<P>The present study aimed to assess anti-inflammatory activity and underlying mechanism of n-propyl gallate, the n-propyl ester of gallic acid. n-Propyl gallate was shown to contain anti-inflammatory activity using two experimental animal models, acetic acid-induced permeability model in mice, and air pouch model in rats. It suppressed production of nitric oxide and induction of inducible nitric oxide synthase and cyclooxygenase-2 in the lipopolysaccharide (LPS)-stimulated RAW264.7 macrophage cells. It was able to diminish reactive oxygen species level elevated in the LPS-stimulated RAW264.7 macrophage cells. It also suppressed gelatinolytic activity of matrix metalloproteinase-9 enhanced in the LPS-stimulated RAW264.7 macrophage cells. It inhibited inhibitory 관B-관 degradation and enhanced NF-관B promoter activity in the stimulated macrophage cells. It was able to suppress phosphorylation of c-Jun NH(2)-terminal kinase 1/2 (JNK1/2) and activation of c-Jun promoter activity in the stimulated macrophage cells. In brief, n-propyl gallate possesses anti-inflammatory activity via down-regulation of NF-관B and JNK pathways.</P>

Expression, Characterization and Regulation of a Saccharomyces cerevisiae Monothiol Glutaredoxin (Grx6) Gene in Schizosaccharomyces pombe

Jae-Hoon Lee,Kyunghoon Kim,Eun-Hee Park,Kisup Ahn,Chang-Jin Lim 한국분자세포생물학회 2007 Molecules and cells Vol.24 No.3

Thermoresistant properties of bacterioferritin comigratory protein against high temperature stress in Schizosaccharomyces pombe

류인왕,이수희,임혜원,안기섭,박광학,사재훈,정경진,임창진,김경훈,Ryu, In Wang,Lee, Su Hee,Lim, Hye-Won,Ahn, Kisup,Park, Kwanghark,Sa, Jae-Hoon,Jeong, Kyung Jin,Lim, Chang-Jin,Kim, Kyunghoon The Microbiological Society of Korea 2016 미생물학회지 Vol.52 No.4

The Schizosaccharomyces pombe structural gene encoding bacterioferritin comigratory protein (BCP) was previously cloned using the shuttle vector pRS316 to generate the BCP-overexpressing plasmid pBCP10. The present work aimed to evaluate the thermoresistant properties of BCP against high temperature stress using the plasmid pBCP10. When the S. pombe cells were grown to the early exponential phase and shifted from $30^{\circ}C$ to $37^{\circ}C$ or $42^{\circ}C$, the S. pombe cells harboring pBCP10 grew significantly more at both $37^{\circ}C$ and $42^{\circ}C$ than the vector control cells. After 6 h of the shifting to higher incubation temperatures, they contained the lower reactive oxygen species (ROS) and nitrite content, an index of nitric oxide (NO), than the vector control cells. After the temperature shifts, total glutathione (GSH) content and total superoxide dismutase (SOD) activities were much higher in the S. pombe cells harboring pBCP10 than in the corresponding vector control cells. Taken together, the S. pombe BCP plays a thermoresistant role which might be based upon its ability both to down-regulate ROS and NO levels and to up-regulate antioxidant components, such as total GSH and SOD, and subsequently to maintain thermal stability. 이전의 연구에서, bacterioferritin comigratory protein (BCP)을 인코딩하는 Schizosaccharomyces pombe의 구조유전자를 shuttle vector인 pRS316에 클로닝하여 BCP 과잉발현 플라즈미드인 pBCP10을 제조한 바 있다. 본 연구에서는, 플라즈미드 pBCP10을 사용하여 고온 스트레스에 대한 BCP의 열저항적 성질을 평가하였다. 대수기의 초기까지 성장시킨 S. pombe 세포의 배양 온도를 $30^{\circ}C$에서 $37^{\circ}C$나 $42^{\circ}C$로 전이시키는 경우, pBCP10 함유 S. pombe 세포가 벡터 대조 세포보다 $37^{\circ}C$와 $42^{\circ}C$ 모두에서 유의하기 더 잘 성장하였다. 높은 배양 온도로 전이한 뒤 6시간에서, pBCP10 함유 S. pombe 세포가 벡터 대조 세포보다 낮은 활성산소종(ROS)과 일산화질소(NO)의 지표로 측정된 아질산염(nitrite) 함량을 갖고 있음이 확인되었다. 온도 전이 뒤에, 총 글루타치온(total glutathione) 함량과 총 수퍼옥사이드 디스뮤타제(superoxide dismutase) 활성은 대응되는 벡터 대조 세포보다 pBCP10 함유 S. pombe 세포에서 현저하게 높다는 사실도 확인되었다. 종합하면, S. pombe BCP는 열저항적 역할을 보유하는 데, 활성산소종과 일산화질소에 대한 하강시키는 활성과 총 글루타치온과 수퍼옥사이드 디스뮤타제 등의 항산화 성분들을 상승시키는 활성, 즉 종합적으로 열안정성을 유지하는 활성에 근거하는 것으로 추정되었다