• Title/Summary/Keyword: RNA2

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Cloning of RNA1 Gene from Saccharomyces cerevisiae (Saccharomyces cerevisiae에서 RNA1 유전자의 클로닝)

  • 송영환;고상석;이영석;강현삼
    • Korean Journal of Microbiology
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    • v.27 no.2
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    • pp.77-84
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    • 1989
  • The temperature sensitive (ts) mutation on RNA1 gene of Saccharomyces cerevisiae prevents growth at restrictive temperature ($36^{\circ}C$) by accumulation of precursor tRNA, rRNA and mRNA (Hutchison et al., 1969; Shiokawa and Pogo, 1974; Hopper et al., 1978). RNA1 gene was cloned by complementation of the temperature sensitive growth defect of an rna1-1 mutant strain and identified by retransformation and concomitant loss of recombinant plasmid on non-selective condition. By deletion mapping, it was found that RNA1 gene resides within 3.5kb of BgII fragment.

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Polymorphism of the Promoter Region of Hsp70 Gene and Its Relationship with the Expression of HSP70mRNA, HSF1mRNA, Bcl-2mrna and Bax-AMrna in Lymphocytes in Peripheral Blood of Heat Shocked Dairy Cows

  • Cai, Yafei;Liu, Qinghua;Xing, Guangdong;Zhou, Lei;Yang, Yuanyuan;Zhang, Lijun;Li, Jing;Wang, Genlin
    • Asian-Australasian Journal of Animal Sciences
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    • v.18 no.5
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    • pp.734-740
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    • 2005
  • The blood samples were collected from dairy cows at the same milking stage. The single-strand conformation polymorphism (PCR-SSCP) method was used to analyze for polymorphism at the 5'flanking region of the hsp70 gene. The mRNA expression levels of HSP70, HSF1, Bcl-2 and Bax-$\alpha$ at different daily-mean-temperature were analyzed by relative quantitative RTPCR. The DNA content, cell phase and the ratio of apoptosis of lymphocytes in peripheral blood of dairy cattle at different daily-meantemperature were determined by FCM. The PCR-SSCP products of primer pair 1 showed polymorphisms and could be divided into four genotypes: aa, ab, ac, cc, with the cis-acting element (CCAAT box) included. Mutations in the hsp70 5'flanking region (468-752 bp) had different effects on mRNA expression of HSP70, HSF1, Bcl-2 and Bax-$\alpha$. The ac genotypic cows showed higher expressions of HSP70mRNA, HSF1mRNA and Bcl-2mRNA/Bax-$\alpha$mRNA and lower ratio of apoptosis. These mutation sites can be used as molecular genetic markers to assist selection for anti-heat stress cows.

Bioinformatic Prediction of SNPs within miRNA Binding Sites of Inflammatory Genes Associated with Gastric Cancer

  • Song, Chuan-Qing;Zhang, Jun-Hui;Shi, Jia-Chen;Cao, Xiao-Qin;Song, Chun-Hua;Hassan, Adil;Wang, Peng;Dai, Li-Ping;Zhang, Jian-Ying;Wang, Kai-Juan
    • Asian Pacific Journal of Cancer Prevention
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    • v.15 no.2
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    • pp.937-943
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    • 2014
  • Polymorphisms in miRNA binding sites have been shown to affect miRNA binding to target genes, resulting in differential mRNA and protein expression and susceptibility to common diseases. Our purpose was to predict SNPs (single nucleotide polymorphisms) within miRNA binding sites of inflammatory genes in relation to gastric cancer. A complete list of SNPs in the 3'UTR regions of all inflammatory genes associated with gastric cancer was obtained from Pubmed. miRNA target prediction databases (MirSNP, Targetscan Human 6.2, PolymiRTS 3.0, miRNASNP 2.0, and Patrocles) were used to predict miRNA target sites. There were 99 SNPs with MAF>0.05 within the miRNA binding sites of 41 genes among 72 inflammation-related genes associated with gastric cancer. NF-${\kappa}B$ and JAK-STAT are the two most important signaling pathways. 47 SNPs of 25 genes with 95 miRNAs were predicted. CCL2 and IL1F5 were found to be the shared target genes of hsa-miRNA-624-3p. Bioinformatic methods could identify a set of SNPs within miRNA binding sites of inflammatory genes, and provide data and direction for subsequent functional verification research.

Silence of LncRNA GAS5 Protects Cardiomyocytes H9c2 against Hypoxic Injury via Sponging miR-142-5p

  • Du, Jian;Yang, Si-Tong;Liu, Jia;Zhang, Ke-Xin;Leng, Ji-Yan
    • Molecules and Cells
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    • v.42 no.5
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    • pp.397-405
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    • 2019
  • The regulatory role of long noncoding RNA (lncRNA) growth arrest-specific transcript 5 (GAS5) in both cancerous and noncancerous cells have been widely reported. This study aimed to evaluate the role of lncRNA GAS5 in heart failure caused by myocardial infarction. We reported that silence of lncRNA GAS5 attenuated hypoxia-triggered cell death, as cell viability was increased and apoptosis rate was decreased. This phenomenon was coupled with the down-regulated expression of p53, Bax and cleaved caspase-3, as well as the up-regulated expression of CyclinD1, CDK4 and Bcl-2. At the meantime, the expression of four heart failure-related miR-NAs was altered when lncRNA GAS5 was silenced (miR-21 and miR-142-5p were up-regulated; miR-30b and miR-93 were down-regulated). RNA immunoprecipitation assay results showed that lncRNA GAS5 worked as a molecular sponge for miR-142-5p. More interestingly, the protective actions of lncRNA GAS5 silence on hypoxia-stimulated cells were attenuated by miR-142-5p suppression. Besides, TP53INP1 was a target gene for miR-142-5p. Silence of lncRNA GAS5 promoted the activation of PI3K/AKT and MEK/ERK signaling pathways in a miR-142-5p-dependent manner. Collectively, this study demonstrated that silence of lncRNA GAS5 protected H9c2 cells against hypoxia-induced injury possibly via sponging miR-142-5p, functionally releasing TP53INP1 mRNA transcripts that are normally targeted by miR-142-5p.

Heterogeneity Analysis of the 16S rRNA Gene Sequences of the Genus Vibrio (Vibrio 속 16S rRNA 유전자 염기서열의 이질성 분석)

  • Ki, Jang-Seu
    • Korean Journal of Microbiology
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    • v.45 no.4
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    • pp.430-434
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    • 2009
  • Bacterial 16S rRNA gene sequences have been widely used for the studies on molecular phylogeny, evolutional history, and molecular detections. Bacterial genomes have multiple rRNA operons, of which gene sequences sometimes are variable. In the present study, heterogeneity of the Vibrio 16S rRNA gene sequences were investigated. Vibrio 16S rRNA sequences were obtained from GenBank databases, considering the completion of gene annotation of Vibrio genome sequences. These included V. cholerae, V. harveyi, V. parahaemolyticus, V. splendidus, and V. vulnificus. Chromosome 1 of the studied Vibrio had 7~10 copies of the 16S rRNA gene, and their intragenomic variations were less than 0.9% dissimilarity (more than 99.1% DNA similarity). Chromosome 2 had none or single 16S rRNA gene. Intragenomic 16S rRNA genotypes were detected at least 5 types (V. vulnificus #CMCP6) to 8 types (V. parahaemolyticus #RIMD 2210633, V. harveyi #ATCC BAA-1116). These suggest that Vibrio has high heterogeneity of the 16S rRNA gene sequences.

Ginsenoside Rh2 reduces m6A RNA methylation in cancer via the KIF26B-SRF positive feedback loop

  • Hu, Chunmei;Yang, Linhan;Wang, Yi;Zhou, Shijie;Luo, Jing;Gu, Yi
    • Journal of Ginseng Research
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    • v.45 no.6
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    • pp.734-743
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    • 2021
  • Background: The underlying mechanisms of the potential tumor-suppressive effects of ginsenoside Rh2 are complex. N6-methyladenosine (m6A) RNA methylation is usually dysregulated in cancer. This study explored the regulatory effect of ginsenoside Rh2 on m6A RNA methylation in cancer. Methods: m6A RNA quantification and gene-specific m6A RIP-qPCR assays were applied to assess total and gene-specific m6A RNA levels. Co-immunoprecipitation, fractionation western blotting, and immunofluorescence staining were performed to detect protein interactions and distribution. QRT-PCR, dual-luciferase, and ChIP-qPCR assays were conducted to check the transcriptional regulation. Results: Ginsenoside Rh2 reduces m6A RNA methylation and KIF26B expression in a dose-dependent manner in some cancers. KIF26B interacts with ZC3H13 and CBLL1 in the cytoplasm of cancer cells and enhances their nuclear distribution. KIF26B inhibition reduces m6A RNA methylation level in cancer cells. SRF bound to the KIF26B promoter and activated its transcription. SRF mRNA m6A abundance significantly decreased upon KIF26B silencing. SRF knockdown suppressed cancer cell proliferation and growth both in vitro and in vivo, the effect of which was partly rescued by KIF26B overexpression. Conclusion: ginsenoside Rh2 reduces m6A RNA methylation via downregulating KIF26B expression in some cancer cells. KIF26B elevates m6A RNA methylation via enhancing ZC3H13/CBLL1 nuclear localization. KIF26B-SRF forms a positive feedback loop facilitating tumor growth.

Circadian Variation of the RNA/DNA Ratios in Marbled Flounder Pleuronectes yokohamae (문치가자미 RNA/DNA의 일주기적 변화)

  • Gwak, Woo-Seok
    • Korean Journal of Ichthyology
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    • v.22 no.4
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    • pp.267-272
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    • 2010
  • Laboratory-reared marbled flounder (Pleuronectes yokohamae) juveniles ($23.2{\pm}0.2mm$ total length; age 89 day) were sampled over a 48-h period to determine any diel patterns in RNA/DNA ratios. RNA/DNA ratios were highest during daytime periods (0800, 1100, 1400, 1700 h) and significantly reduced at night (2000, 2300, 0200, 0500 h). Findings from this study indicate a diel variation in biochemical condition and suggest that special care should be taken in designing sampling plans, including sampling time and data analysis to account for this source of variability.

Recent Progress of Structural Biology of tRNA Processing and Modification

  • Nakanishi, Kotaro;Nureki, Osamu
    • Molecules and Cells
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    • v.19 no.2
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    • pp.157-166
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    • 2005
  • Transfer RNA (tRNA) is a key molecule to decode the genetic information on mRNA to amino aicds (protein), in a ribosome. For tRNA to fulfill its adopter function, tRNA should be processed into the standard length, and be post-transcriptionally modified. This modification step is essential for the tRNA to maintain the canonical L-shaped structure, which is required for the decoding function of tRNA. Otherwise, it has recently been proposed that modification procedure itself contributes to the RNA (re)folding, where the modification enzymes function as a kind of RNA chaperones. Recent genome analyses and post-genome (proteomics and transcriptomics) analyses have identified genes involved in the tRNA processings and modifications. Furthermore, post-genomic structural analysis has elucidated the structural basis for the tRNA maturation mechanism. In this paper, the recent progress of the structural biology of the tRNA processing and modification is reviewed.

Single-molecule fluorescence measurements reveal the reaction mechanisms of the core-RISC, composed of human Argonaute 2 and a guide RNA

  • Jo, Myung Hyun;Song, Ji-Joon;Hohng, Sungchul
    • BMB Reports
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    • v.48 no.12
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    • pp.643-644
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    • 2015
  • In eukaryotes, small RNAs play important roles in both gene regulation and resistance to viral infection. Argonaute proteins have been identified as a key component of the effector complexes of various RNA-silencing pathways, but the mechanistic roles of Argonaute proteins in these pathways are not clearly understood. To address this question, we performed single-molecule fluorescence experiments using an RNA-induced silencing complex (core-RISC) composed of a small RNA and human Argonaute 2. We found that target binding of core-RISC starts at the seed region of the guide RNA. After target binding, four distinct reactions followed: target cleavage, transient binding, stable binding, and Argonaute unloading. Target cleavage required extensive sequence complementarity and accelerated core-RISC dissociation for recycling. In contrast, the stable binding of core-RISC to target RNAs required seed-match only, suggesting a potential explanation for the seed-match rule of microRNA (miRNA) target selection.

Paramyxoviridae -Classification, structure, genomes and their encoded proteins- (파라믹소바이러스 -분류, 구조, 게놈 및 코드화된 단백질-)

  • 송희종;어성국;채효석
    • Korean Journal of Veterinary Service
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    • v.25 no.2
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    • pp.153-174
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    • 2002
  • 파라믹소바이러스과 (Parnxouiridae) 바이러스는 외피당단백질의 생물학적 특성에 있어서 Orthomyxoviridae와, 분절되지 않은(nonseg-mented)게놈과 그 발현의 구성 측면에서 Rhbdoviridae와 유사성을 갖는 바이러스로서 외피로 둘러 싸여 있는 마이너스쇄(negative strand) RNA 바이러스이다. 마이너스쇄 RNA 바이러스의 게놈 RNA는 2가지 기능으로 작용한 다. 즉, 첫째는 mRNA 합성을 위한 주형(鑄型, template)으로 작용하고, 둘째는 플러스쇄 (anti-genome(+) strand)의 합성을 위한 주형으로서 작용한다. 마이너스쇄 RNA바이러스는 그들 자신의 RNA전사효소를 코드화 하여 저장하지만, mRNA는 감염세포 내에서 게놈이 노출된 후에만 합성된다. 바이러스 복제는 mRNA의 합성후 일어나며 연속된 바이러스단백질의 합성을 필요로 한다. 새로 합성된 플러스쇄(antigenome(+)strand)는 마이너스쇄 게놈 RNA의 복제를 도모하기 위한 주형(鑄型)으로서 작용한다.