• Korean Journal of Microbiology
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• v.47 no.4
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• pp.289-294
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• 2011

We investigated the cold shock sensitivity of DEAD-box RNA helicase gene deleted strains of in Bacillus subtilis CU1065. To understand cold shock effects, cells were cultivated at $37^{\circ}C$ to log phase ($O.D_{600}$=0.5-0.6) and then temperature was shifted to $15^{\circ}C$. Cold shock slow down the growth rate of wild type and deleted strains of DEAD-box RNA helicase gene (ydbR, yfmL, yqfR, deaD). The growth rate of ydbR deleted strain is 5 times severely reduced compared to that of wild type strain (CU1065). But the growth rate of other three (yfmL, yqfR, deaD) deleted strains is nearly equal to the growth rate of wild type. Compared to $37^{\circ}C$, the amount of ydbR and yqfR mRNA transcripts are increased at the growth temperature of $15^{\circ}C$. On the other hands the mRNA transcripts of yfmL and deaD are not changed at both conditions of $37^{\circ}C$ and $15^{\circ}C$. Upon cold shock treatment ydbR mRNA transcript is clearly increased. After treatment of rifampicin (bacteria transcription inhibitor) the amount of ydbR mRNA was measured. Temperature shift from $37^{\circ}C$ to $15^{\circ}C$ and rifampicin treatment showed slowly decay of ydbR mRNA. But at $37^{\circ}C$ and rifampicin treatment ydbR mRNA is rapidly reduced. These results showed that cold shock induction of ydbR mRNA resulted from the stability of ydbR mRNA and not from the transcription induction of ydbR. In relation to these results, we found the cold box element of csp (cold shock protein gene) in 5' untranslated region of ydbR gene. Cold shock induction of ydbR is caused by the stability of ydbR mRNA like the stability of csp mRNA.

• Journal of Aquaculture
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• v.20 no.1
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• pp.65-72
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• 2007

Full length complementary DNA encoding apolipoprotein A-I (apoA-I) was isolated and characterized in mud loach (Misgurnus mizolepis). Mud loach apoA-I cDNA encoding 24 bp of 5'-untranslated region (UTR), 762 bp of single open reading frame (ORF) consists of 254 amino acids and 293 bp of 3'-UTR excluding stop codon and poly (A+) tail. Two overlapping polyadenylation signals (AATAAAATAAA) was found 9 bp prior to the poly (A+) tail. Mud loach apoA-I represented considerable homology to those from other teleost species at amino acid level with conserving common features of vertebrate apoA-I. Molecular phylogenetic analysis inferred the phylogenetic hypothesis that was generally in accordance with the previous taxonomic relationship. Apolipoprotein A-I mRNA was detected in various tissues, but the mRNA levels were quite varied depending on tissues based on semi-quantitative RT-PCR. Liver and brain showed the significantly higher levels of apoA-I transcripts than other tissues. mRNA expression of apoA-I was quite low in very early stage of embryonic development, however dramatically enhanced from 8 hours post fertilization. This increased mRNA level was retained consistently up to 14 days post hatching.

• Molecules and Cells
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• v.45 no.7
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• pp.465-478
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• 2022

MicroRNAs (miRNAs) are a class of small non-coding RNAs that regulate the expression of target messenger RNA (mRNA) complementary to the 3' untranslated region (UTR) at the post-transcriptional level. Hsa-miR-422a, which is commonly known as miRNA derived from transposable element (MDTE), was derived from short interspersed nuclear element (SINE). Through expression analysis, hsa-miR-422a was found to be highly expressed in both the small intestine and liver of crab-eating monkey. AT-Rich Interaction Domain 5 B (ARID5B) was selected as the target gene of hsa-miR-422a, which has two binding sites in both the exon and 3'UTR of ARID5B. To identify the interaction between hsa-miR-422a and ARID5B, a dual luciferase assay was conducted in HepG2 cell line. The luciferase activity of cells treated with the hsa-miR-422a mimic was upregulated and inversely downregulated when both the hsa-miR-422a mimic and inhibitor were administered. Nuclear factor erythroid-2 (NF-E2) was selected as the core transcription factor (TF) via feed forward loop analysis. The luciferase expression was downregulated when both the hsa-miR-422a mimic and siRNA of NF-E2 were treated, compared to the treatment of the hsa-miR-422a mimic alone. The present study suggests that hsa-miR-422a derived from SINE could bind to the exon region as well as the 3'UTR of ARID5B. Additionally, hsa-miR-422a was found to share binding sites in ARID5B with several TFs, including NF-E2. The hsa-miR-422a might thus interact with TF to regulate the expression of ARID5B, as demonstrated experimentally. Altogether, hsa-miR-422a acts as a super enhancer miRNA of ARID5B by collaborating with TF and NF-E2.

• Journal of Life Science
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• v.33 no.12
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• pp.987-994
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• 2023

This study examined the associations between the genotypes of the galactose mutarotase (GALM) gene and carcass traits in the Hanwoo population of Jeju Island, South Korea. The GALM genotypes were determined by the 14-bp (5'-GGTCTAATGACCAG-3') insertion/deletion (InDel) polymorphisms of the 3'-untranslated region (UTR). All three genotypes (LL, LS, and SS) were found in the Hanwoo steer population. The association analysis showed significant associations between genotypes and several carcass traits, including traits related to intramuscular fat content, such as meat quality, marbling score, and backfat thickness (p<0.05). Animals harboring the SS genotype showed not only higher levels of intramuscular fat content but also lower levels of backfat thickness than animals harboring the LL and LS genotypes. On the other hand, no significant associations were found between the GALM genotypes and carcass weight, eye muscle area, meat color, or fat color (p>0.05). Deleting the 14-bp segment in the 3'-UTR resulted in the modification of the secondary structure of RNA and appeared to affect gene expression by interfering with the binding ability of GALM mRNA with RNA-binding proteins and microRNAs. These results suggest that the 14-bp InDel polymorphism in the 3'-UTR region of the GALM gene affects cattle growth traits and carcass quality through galactose metabolism-mediated fat accumulation in muscle and backfat tissues.

• BMB Reports
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• v.51 no.11
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• pp.602-607
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• 2018

Aberrant expression of microRNAs (miRNAs) plays important roles in carcinogenesis and tumor progression. However, the expression and biological role of miR-301b in triple-negative breast cancer (TNBC) remains unclear. Here we aimed to evaluate the roles and mechanisms of miR-301b in TNBC cells. miR-301b expression was assessed in TNBC specimens and cell lines by quantitative Real-Time PCR (qRT-PCR). TNBC cells were transfected with miR-301b mimics, inhibitors or Cylindromatosis (CYLD) small interfering RNA (siRNA) using Lipofectamine 2000. The functional roles of miR-301b were determined by cell proliferation, colony formation, and apoptosis assays. Western blots and qRT-PCR were used to measure the expression of mRNAs and proteins in the cells. We found that miR-301b was upregulated in TNBC specimens and cell lines. Overexpression of miR-301b promoted cell proliferation in TNBC cells, while inhibited the apoptosis induced by 5-FU. CYLD was downregulated by miR-301b at both mRNA and protein levels in TNBC cells. Dual-luciferase report assay confirmed that miR-301b downregulated CYLD by direct interaction with the 3'-untranslated region(3'-UTR) of CYLD mRNA. $NF-{\kappa}B$ activation was mechanistically associated with miR-301b-mediated downregulation of CYLD. However, inhibition of miR-301b reversed all the effects of miR-301b. In conclusion, miR-301b plays an oncogenic role in TNBC possibly by downregulating CYLD and subsequently activating $NF-{\kappa}B$ p65, and this may provide a novel therapeutic approach for TNBC.

• Asian Pacific Journal of Cancer Prevention
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• v.15 no.16
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• pp.6949-6954
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• 2014

Background: Recent studies have indicated that microRNA-15a (miR-15a) is dysregulated in breast cancer (BC). We aimed to evaluate the expression of miR-15a in BC tissues and corresponding para-carcinoma tissues. We also focused on effects of miR-15a on cellular behavior of MDA-MB-231 and expression of its target gene synuclein-${\gamma}$ (SNCG). Materials and Methods: The expression levels of miR-15a were analysed in BC formalin fixed paraffin embedded (FFPE) tissues by microarray and quantitative real-time PCR. CCK-8 assays, cell cycle and apoptosis assays were used to explore the potential functions of miR-15a in MDA-MB-231 human BC cells. A luciferase reporter assay confirmed direct targets. Results: Downregulation of miR-15a was detected in most primary BCs. Ectopic expression of miR-15a promoted proliferation and suppressed apoptosis in vivo. Further studies indicated that miR-15a may directly interact with the 3'-untranslated region (3'-UTR) of SNCG mRNA, downregulating its mRNA and protein expression levels. SNCG expression was negatively correlated with miR-15a expression. Conclusions: MiR-15a has a critical role in mediating cell cycle arrest and promoting cell apoptosis of BC, probably by directly targeting SNCG. Thus, it may be involved in development and progression of BC.

• Journal of the Korean Society of Food Science and Nutrition
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• v.35 no.5
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• pp.661-670
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• 2006

Selenium (Se) obtained from dietary sources including cereals, grains and vegetables is an essential micronutrient for normal function of the body. Plants convert Se into selenomethionine and incorporate it into proteins in place of methionine, while higher animals synthesize selenoproteins containing selenocysteine. Excessive Se in the body is methylated stepwise to methylated selenium metabolites from selenide. Both inorganic and organic forms of selenium can be the nutritional sources in human, and they are transformed to selenide and then the amino acid selenocysteine attached to a specific $tRNA^{ser(sec)}$. The selenocysteine (Sec) is incorporated into selenoprotein sequences by the UGA codon. The decoding of UGA as Sec requires specific mechanisms because UGA is normally read as a stop codon: cis-acting sequences in the mRNA (the selenocysteine insertion sequence, SECIS, within the 3'untranslated region) and trans -acting factors dedicated to Sec incorporation are required for incorporation of Sec during translation of selenoprotein mRNAs. Approximately 25 selenoproteins have been identified in mammals. Several of these, including glutathione peroxidases, thioredoxin reductases and selenoprotein P, have been purified or cloned, allowing further characterization of their biological function. The antioxidant properties of selenoproteins help prevent cellular damage from free radicals which may contribute to the development of chronic disease such as cancer and heart disease. Other selenoproteins have important roles in regulation of thyroid function and play a role in the immune system. Daily selenium iatake was reported to be $42.0{\pm}16.9{\mu}g/day$ in Korean adult women. This review focuses on the metabolism and biological functions of selenium, and the nutritional status of selenium in the Korean population.

• Development and Reproduction
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• v.14 no.1
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• pp.13-19
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• 2010

We developed a novel dicistronic system for the expression of target cDNA sequences in the milk of transgenic animals using goat beta-casein/hGH fusion construct, pGbc5.5hGH (Lee, 2006) and internal ribosome entry site (IRES) sequences of encephalomyocarditis virus (EMCV). Granulocyte colony-stimulating factor (hG-CSF) cDNA was linked to 3' untranslated region of hGH gene in the pGbc5.5hGH via EMCV IRES sequences. Transgenic mice were generated by microinjection and transgene expression was examined in the milk and mammary gland of transgenic mice at 10 days of lactation. Northern blot analysis showed that hGH gene and hG-CSF cDNA were transcribed as a single dicistronic mRNA. The hG-CSF and hGH proteins were independently translated from the dicistronic mRNA and secreted into the milk of transgenic mice. The highest concentration of hG-CSF and hGH in the milk of transgenic mice were $237{\mu}g/m{\ell}$ and $8,990{\mu}g/m{\ell}$, respectively. In contrast, another hG-CSF expression cassette, in which hG-CSF genomic sequences were inserted into a commercial milk-specific expression vector (pBC1), generated a lower level ($91{\mu}g/m{\ell}$) of hG-CSF expression in the milk of transgenic mice. These results demonstrated that the novel pGbc5.5hGH-based dicistronic construct could be useful for an efficient cDNA expression in the milk of transgenic animals.

• BMB Reports
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• v.50 no.4
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• pp.208-213
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• 2017

Vascular endothelial growth factor (VEGF) is an essential cytokine that has functions in the formation of new blood vessels and regression of cardiac hypertrophy. VEGF/VEGF-receptor-1 (VEGFR1) signaling plays a key role in the regression of cardiac hypertrophy, whereas VEGF/VEGFR2 signaling leads to cardiac hypertrophy. In this study, we identified the prohypertrophic role of miR-374 using neonatal rat ventricular myocytes (NRVMs). Our results showed that overexpression of miR-374 activated G protein-coupled receptor-mediated prohypertrophic pathways by the inhibition of VEGFR1-dependent regression pathways. Luciferase assays revealed that miR-374 could directly target the 3'-untranslated regions of VEGFR1 and cGMP-dependent protein kinase-1. Collectively, these findings demonstrated that miR-374 was a novel pro-hypertrophic microRNA functioning to suppress the VEGFR1-mediated regression pathway.

• Asian Pacific Journal of Cancer Prevention
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• v.14 no.9
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• pp.5017-5021
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• 2013

Objective: MicroRNAs (miRNAs) are a small class of non-coding, single-stranded RNAs with a critical role in genesis and maintenance of renal cancer mainly through binding to 3'-untranslated regions (3'UTR) of target mRNAs, which causes a block of translation and/or mRNA degradation. The aim of the present study was to investigate the potential effects of miR-122 in human renal cell carcinomas. Methods: The expression level of miR-122 was quantified by qRT-PCR. MTT, colony formation, invasion and migration assays were used to explore the potential functions of miR-122 in human renal cell carcinoma cells. Results: Cellular growth, invasion and migration in two A498 and 786-O cells were significantly increased after miR-122 transfection. Further experiments demonstrated that overexpression of miR-122 resulted in the increase of phospho-Akt (Ser473) and phospho-mTOR (Ser2448), then activation of mTOR targets, p70S6K and 4E-BP1. Conclusions: The up-regulation of miR-122 may play an important role in the progress of renal cancer through activating PI3K/Akt signal pathway and could be a potential molecular target for anti-cancer therapeutics.