• Bulletin of the Korean Chemical Society
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• v.22 no.5
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• pp.514-518
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• 2001

We have explored the importance of two ATP binding domains of Hsp104 protein in protection of yeast cells from cadmium exposure. In the previous study we have discovered that the presence of two ATP binding sites was essential in providing heat sh ock protection as well as rescuing cells from oxidative stress. In this paper we first report wild type cell with functional hsp104 gene is more resistant to cadmium stress than hsp104-deleted mutant cell, judging from decrease in survival rates as a result of cadmium exposure. In order to demonstrate functional role of two ATP binding sites in cadmium defense, we have transformed both wild type (SP1) and hyperactivated ras mutant (IR2.5) strains with several plasmids differing in the presence of ATP binding sites. When an extra copy of functional hsp104 gene with both ATP binding sites was overexpressed with GPD-promoter, cells showed increased survival rate against cadmium stress than mutants with ATP binding sites changed. The degree of protection in the presence of two ATP binding sites was similarly observed in ira2-deleted hyperactivated ras mutant, which was more sensitive to oxidative stress than wild type cell. We have concluded that the greater sensitivity to cadmium stress in the absence of two ATP binding sites is attributed to the higher concentration of reactive oxygen species (ROS) produced by cadmium exposure based on the fluorescence tests. These findings, taken all together, imply that the mechanism by which cadmium put forth toxic effects may be closely associated with the oxidative stress, which is regulated independently of the Ras-cAMP pathway. Our study provides a better understanding of cadmium defense itself and cross-talks between oxidative stress and metal stress, which can be applied to control human diseases due to similar toxic environments.

• Toxicological Research
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• v.22 no.4
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• pp.323-328
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• 2006

Global gene expression profile was analyzed by microarray analysis of rat liver RNA after acute acetaminophen (APAP) administration. A single dose of 1g/kg body weight of APAP was given orally, and the liver samples were obtained after 24, 48 h, and 2 weeks. Histopathologic and biochemical studies enabled the classification of the APAP effect into injury (24 and 48 h) and regeneration (2 weeks) stages. The expression levels of 4900 clones on a custom rat gene microarray were analyzed and 484 clones were differentially expressed with more than a 1.625-fold difference(which equals 0.7 in log2 scale) at one or more time points. Two hundred ninety seven clones were classified as injury-specific clones, while 149 clones as regeneration-specific ones. Characteristic gene expression profiles could be associated with APAP-induced gene expression changes in lipid metabolism, stress response, and protein metabolism. We established a global gene expression profile utilizing microarray analysis in rat liver upon acute APAP administration with a full chronological profile that not only covers injury stage but also later point of regeneration stage.

• BMB Reports
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• v.38 no.4
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• pp.432-439
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• 2005

$Ca^{2+}$/calmodulin transduction pathways have been implicated in mediating stress response and tolerance in plants. Here, three genes encoding calmodulin (Cam) members of the EF-hand family of $Ca^{2+}$-binding proteins were identified from Oryza sativa L. databases. Complementary DNA for each of the calmodulin genes, OsCam1, OsCam2, and OsCam3 were sequenced. OsCam1 and OsCam2 encode a conventional 148-amino acid calmodulin protein that contains four characteristic $Ca^{2+}$-binding motifs. OsCam3 encode a similar protein with a 38-amino-acid extension containing a putative prenylation site (CVIL) at the carboxyl terminus. RT-PCR showed that each of the genes is expressed in leaves and roots of 2-week old rice seedlings. By RNA gel blot analysis, OsCam1 mRNA levels strongly increased in response to NaCl, mannitol and wounding treatments. In contrast, OsCam2 mRNA levels were relatively unchanged under all conditions investigated. NaCl treatment and wounding also increased the OsCam3 mRNA level, but in a more transient manner. Our results indicate that although the expression of genes encoding different calmodulin isoforms is ubiquitous, they are differentially regulated by various stress signals. In addition, we have demonstrated that the calcium-channel blocker lanthanum chloride inhibited the induction of OsCam1 gene expression by both NaCl and mannitol treatments. These results suggest that osmotic stress induced expression of OsCam1 gene requires the $[Ca^{2+}]_{cyt}$ elevation that is known to occur in response to these stimuli.

• Journal of Microbiology and Biotechnology
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• v.17 no.9
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• pp.1504-1512
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• 2007

The fpr gene, which encodes a ferredoxin-$NADP^+$ reductase, is known to participate in the reversible redox reactions between $NADP^+$/NADPH and electron carriers, such as ferredoxin or flavodoxin. The role of Fpr and its regulatory protein, FinR, in Pseudomonas putida KT2440 on the oxidative and osmotic stress responses has already been characterized [Lee at al. (2006). Biochem. Biophys. Res. Commun. 339, 1246-1254]. In the genome of P. putida KT2440, another Fpr homolog (FprB) has a 35.3% amino acid identity with Fpr. The fprB gene was cloned and expressed in Escherichia coli. The diaphorase activity assay was conducted using purified FprB to identify the function of FprB. In contrast to the fpr gene, the induction of fprB was not affected by oxidative stress agents, such as paraquat, menadione, $H_2O_2$, and t-butyl hydroperoxide. However, a higher level of fprB induction was observed under osmotic stress. Targeted disruption of fprB by homologous recombination resulted in a growth defect under high osmotic conditions. Recovery of oxidatively damaged aconitase activity was faster for the fprB mutant than for the fpr mutant, yet still slower than that for the wild type. Therefore, these data suggest that the catalytic function of FprB may have evolved to augment the function of Fpr in P. putida KT2440.

• Journal of Microbiology and Biotechnology
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• v.17 no.4
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• pp.586-593
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• 2007

Seed coating by a phenazine-producing bacterium, Pseudomonas chlororaphis O6, induced dose-dependent inhibition of germination in wheat and barley seeds, but did not inhibit germination of rice or cucumber seeds. In wheat seedlings grown from inoculated seeds, phenazine production levels near the seed were higher than in the roots. Deletion of the gacS gene reduced transcription from the genes required for phenazine synthesis, the regulatory phzI gene and the biosynthetic phzA gene. The inhibition of seed germination and the induction of systemic disease resistance against a bacterial soft-rot pathogen, Erwinia carotovora subsp. carotovora, were impaired in the gacS and phzA mutants of P chlororaphis O6. Culture filtrates of the gacS and phzA mutants of P. chlororaphis O6 did not inhibit seed germination of wheat, whereas that of the wild-type was inhibitory. Our results showed that the production of phenazines by P. chlororaphis O6 was correlated with reduced germination of barley and wheat seeds, and the level of systemic resistance in tobacco against E. carotovora.

• Journal of Life Science
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• v.11 no.5
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• pp.464-469
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• 2001

A gene, dnaK2, encoding a distinct member of the HSP70 family of molecular chaperones is isolated from the halotolerant cyanobactrium Aphanothece halophytica. The dnak2 gene encodes a molecular wight of 68 kDa polypeptide with predicted 616 amino acid residues. The DnaK2 protein has a structural characteristic of bacterial DnaK homologues and shows high similarity to other HSP70/Dank proteins. The danK2 transcripts are hardly detectable at 28$^{\circ}C$ and strongly induced upon heat stress. It is also found that dnaK2 transcript is increased by high-salinity stress even in the absence of heat stress. These results suggest that the DnaK2 protein plays an important role in protecting A. halophytica against damage caused by salt stress at well as heat stress.

• Korean Journal of Plant Resources
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• v.23 no.1
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• pp.25-30
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• 2010

This study was carried out to understand the effect of low temperature($4^{\circ}C$), heat shock($37^{\circ}C$) and drought stresses on the growth and gene expression of Arabidopsis ATSIZ3(at1g08910) mutants. The seedling growth of SIZ3-mutants were markedly inhibited by the treatment of heat shock or chilling stresses. However, there was no significant differences between wild type and SIZ3-mutants in seeding fresh weight. As compared to wild type plants, SIZ3-mutants showed 63.9% inhibition of seedling fresh weight by the treatment of 10 days drought stress, suggesting that SIZ3 is involved in the resistance of Arabidopsis to drought stress. Base on RT-PCR analysis, expression of SIZ3 mRNA in the wild type showed 20% inhibition by chilling stress, 3.7 and 4.5 fold increase by the treatment of heat shock or drought stresses, respectively.

• The Plant Pathology Journal
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• v.35 no.5
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• pp.445-458
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• 2019

The lipopolysaccharide (LPS) composed of lipid A, core, and O-antigen is the fundamental constituent of the outer membrane in gram-negative bacteria. This study was conducted to investigate the roles of LPS in Burkholderia glumae, the phytopathogen causing bacterial panicle blight and seedling rot in rice. To study the roles of the core oligosaccharide (OS) and the O-antigen region, mutant strains targeting the waaC and the wbiFGHI genes were generated. The LPS profile was greatly affected by disruption of the waaC gene and slight reductions were observed in the O-antigen region following wbiFGHI deletions. The results indicated that disruption in the core OS biosynthesis-related gene, waaC, was associated with increased sensitivity to environmental stress conditions including acidic, osmotic, saline, and detergent stress, and to polymyxin B. Moreover, significant impairment in the swimming and swarming motility and attenuation of bacterial virulence to rice were also observed in the waaC-defective mutant. The motility and virulence of O-antigen mutants defective in any gene of the wbiFGHI operon, were not significantly different from the wild-type except in slight decrease in swimming and swarming motility with wbiH deletion. Altogether, the results of present study indicated that the LPS, particularly the core OS region, is required for tolerance to environmental stress and full virulence in B. glumae. To our knowledge, this is the first functional study of LPS in a plant pathogenic Burkholderia sp. and presents a step forward toward full understanding of B. glumae pathogenesis.

• Proceedings of the Korean Society of Plant Pathology Conference
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• 2003.10a
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• pp.81.1-81
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• 2003

Although plants have evolved to possess various defense mechanisms from local biotic and abiotic stressors, most of yield loss is caused by theses stressors. Recent studies have revealed that several different stress responsive reactions are inter-networking. Therefore, the identification and dissection of stress responsive genes is an essential and first step towards understanding of the global defense mechanism in response to various stressors. For this purpose, we applied cDNA microarray analysis, because it has powerful ability to monitor the global gene expression in a specific situation. To date, more than 10,000 non-redundant genes were identified from seven different cDNA libraries and deposited in our EST database (http://plant.pdrs.re.kr/ks200201/pepper.html). For this study, we have built 5K cDNA microarray containing 4,685 unigene clones from three different cDNA libraries. Monitoring of gene expression profiles of hot pepper interactions with biotic stress, abiotic stresses and chemical treatments will be presented. Although this work shows expression profiling at the sub-genomic level, this could be a good starting point to understand the complexity of global defense mechanism in hot pepper.

• Journal of Life Science
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• v.11 no.1
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• pp.30-33
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• 2001

To identify novel components involved in the salt stress signaling pathway of yeast cells, we used mTn3-mediated transposon tagging library and screened mutants displaying enhanced tolerance to NaCl. Southern blot analysis indicated that more than 80% of the sre (salt resistant) mutants possessed only one insertion of the tagged transposon, suggesting that the NaCl resistant phenotype was mediated by a single gene in the majority of the mutants. To define the role of SRE genes in the salt stress signaling pathway, we introduced NaCl stress-inducible ENA1::LacZ construct into the sre mutants and examined the expression of ${\beta}$-galactosidase activity. Interestingly, we could detect high level of ${\beta}$-galactosidase activity without any NaCl treatment in the sre-3, 4, 6 and 7 mutants. These results indicate that SRE-3, 4, and 7 gene are components of salt stress signaling pathway of yeast cells.