• Title/Summary/Keyword: RNA secondary structure

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Phylogenetic Study of Ganoderma applanatum and Schizopora paradoxa Basd on 5S rRNA Sequences (5S rRNA 염기서열에 으한 잔나비걸상과 좀구멍버섯의 계통학적 연구)

  • Kim, Hak-Hyun;Jung, Hack-Sung
    • Korean Journal of Microbiology
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    • v.32 no.3
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    • pp.177-181
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    • 1994
  • The sequences of the cytoplasmic 5S rRNAs(EMBL accession number X73589 and X73890) from two polupores, Ganoderma applanatum and Schizopora paradoxa, were determined by the direct chemical method for sequencing RNA and compared to the sequences of 9 reported mushrooms. 5S rRNAs of Ganoderma applanatum and Schizopora paradoxa consisted of 118 bases and fit the secondary structure model of the 5S rRNAs of basidiomycetes proposed by Huysmans et al. Based on Kimura’s K_nuc values, the closest fungus to Ganoderma applanatum was Ceratobasidium cornigerum and the one to Schizopora paradoxa was Bjerkandera adusta. When the secondary structures of 5S rRNAs of 11 mushrooms were compared the base substitution occurred at helix regions more than at loop regions. When a phylogenetic tree was constructed using the Neighbor program of the PHYLIP package, it partially discriminated and separated the mushrooms of the Hymenomycetes by the order.

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Application of multi dimensional NMR experiments to VBS RNAs of Yeast Saccaromyces cerevisiae virus

  • Chaejoon Cheong;Cheong, Hae-Kap;Yoo, Jun-Seok
    • Journal of the Korean Magnetic Resonance Society
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    • v.5 no.1
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    • pp.29-36
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    • 2001
  • The structures of two VBS (viral binding site) RNAs, SL1 and SL2, of Yeast Saccharomyces cerevisiae vims have been studied by 2D and 3D NMR experiments. VBSs play a crucial role in viral particle binding to the plus strand and packaging of the RNA. The secondary structures of the two VBS RNAs share a common feature of the stem-internal loop-stem-hairpin loop structure although the size of the internal loops of SL1 and SL2 differs. 2D experiments were sufficient for fill assignments of SL1. However, isotope labeling of the sample and multidimensional experiments were required for 28-nucleotide-long SL2 due to the spectral overlap. Several 3D HCCH experiments have accomplished full assignment of SL2 RNA.

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NMR Study of Effects of $MgCl_2$ on the Structural and Dynamical Properties of Yeast Phenylalanyl tRNA

  • Se Won Suh;Byong Seok Choi;Ki Hang Choi;Jin Young Park
    • Bulletin of the Korean Chemical Society
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    • v.13 no.5
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    • pp.517-520
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    • 1992
  • Solvent exchange rates of selected protons were measured by NMR saturation recovery method for yeast $tRNA^{Phe}$, at temperature from 25 to $40^{\circ}C$, in the presence of 0.1 M NaCl and various low levels of added magnesium ion. The exchange rates in zero $Mg^{2+}$ concentration indicate early melting of acceptor stem, D stem, and tertiary structure. Addition of magnesium ion stabilizes the entire D stem more effectively than any other secondary or tertiary interactions.

1H, 15N, and 13C Resonance Assignments of the Anti-CRISPR AcrIIA4 from Listeria monocytogenes Prophages

  • Kim, Iktae;Kim, Nak-Kyoon;Suh, Jeong-Yong
    • Journal of the Korean Magnetic Resonance Society
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    • v.22 no.3
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    • pp.71-75
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    • 2018
  • The CRISPR-Cas system is the adaptive immune system in bacteria and archaea against invading phages or foreign plasmids. In the type II CRISPR-Cas system, an endonuclease Cas9 cleaves DNA targets of phages as directed by guide RNA comprising crRNA and tracrRNA. To avoid targeting and destruction by Cas9, phages employ anti-CRISPR (Acr) proteins that act against host bacterial immunity by inactivating the CRISPR-Cas system. Here we report the backbone $^1H$, $^{15}N$, and $^{13}C$ resonance assignments of AcrIIA4 that inhibits endonuclease activity of type II-A Listeria monocytogenes Cas9 and also Streptococcus pyogenesis Cas9 using triple resonance nuclear magnetic resonance spectroscopy. The secondary structures of AcrIIA4 predicted by the backbone chemical shifts show an ${\alpha}{\beta}{\beta}{\beta}{\alpha}{\alpha}$ fold, which is used to determine the solution structure.

Development and Evaluation of a SYBR Green Real-time PCR Assay for Canine Cytokine Gene Expression (SYBR Green 실시간 역전사 중합효소연쇄반응을 이용한 개 싸이토카인 유전자 발현의 정량)

  • Yu, Do-Hyeon;Ihn, Dong-Chul;Park, Chul;Park, Jin-Ho
    • Journal of Veterinary Clinics
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    • v.27 no.5
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    • pp.508-513
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    • 2010
  • Cytokines are important mediators of the immune response, and quantitating cytokine mRNA is a highly sensitive and attractive method for measuring cytokine production. The objective of the current study was to develop and validate a SYBR green quantitative real-time reverse transcriptase PCR (qRT-PCR) assay for measuring canine cytokine mRNA. The optimal annealing temperatures ($T_a$) of the designed primers were $62^{\circ}C$ for interleukin (IL)-$1{\beta}$, IL-6 and IL-10; $60^{\circ}C$ for glyceraldehyde 3-phosphate dehydrogenase (GAPDH) and tumor necrosis factor (TNF)-${\alpha}$; and $58^{\circ}C$ for high mobility group box 1 (HMGB1). Primer efficiencies of all primers calculated for standard curve samples were between 97.1% and 102.6%. No evidence of secondary structure or primer-dimer formation was seen via melt-curve analysis or gel electrophoresis. The developed qRT-PCR assays are highly specific and sensitive and can be used to quantify gene expression levels of canine cytokines.

Molecular characterization and expression pattern of a novel Keratin-associated protein 11.1 gene in the Liaoning cashmere goat (Capra hircus)

  • Jin, Mei;Cao, Qian;Wang, Ruilong;Piao, Jun;Zhao, Fengqin;Piao, Jing'ai
    • Asian-Australasian Journal of Animal Sciences
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    • v.30 no.3
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    • pp.328-337
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    • 2017
  • Objective: An experiment was conducted to determine the relationship between the KAP11.1 and the regulation wool fineness. Methods: In previous work, we constructed a skin cDNA library and isolated a full-length cDNA clone termed KAP11.1. On this basis, we conducted a series of bioinformatics analysis. Tissue distribution of KAP11.1 mRNA was performed using semi-quantitative reverse transcription polymerase chain reaction (RT-PCR) analysis. The expression of KAP11.1 mRNA in primary and secondary hair follicles was performed using real-time PCR (real-time polymerase chain reaction) analysis. The expression location of KAP11.1 mRNA in primary and secondary hair follicles was performed using in situ hybridization. Results: Bioinformatics analysis showed that KAP11.1 gene encodes a putative 158 amino acid protein that exhibited a high content of cysteine, serine, threonine, and valine and has a pubertal mammary gland) structural domain. Secondary structure prediction revealed a high proportion of random coils (76.73%). Semi-quantitative RT-PCR showed that KAP11.1 gene was expressed in heart, skin, and liver, but not expressed in spleen, lung and kidney. Real time PCR results showed that the expression of KAP11.1 has a higher expression in catagen than in anagen in the primary hair follicles. However, in the secondary hair follicles, KAP11.1 has a significantly higher expression in anagen than in catagen. Moreover, KAP11.1 gene has a strong expression in inner root sheath, hair matrix, and a lower expression in hair bulb. Conclusion: We conclude that KAP11.1 gene may play an important role in regulating the fiber diameter.

A Modeling Study of Co-transcriptional Metabolism of hnRNP Using FMR1 Gene

  • Ro-Choi, Tae Suk;Choi, Yong Chun
    • Molecules and Cells
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    • v.23 no.2
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    • pp.228-238
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    • 2007
  • Since molecular structure of hnRNP is not available in foreseeable future, it is best to construct a working model for hnRNP structure. A geometric problem, assembly of $700{\pm}20$ nucleotides with 48 proteins, is visualized by a frame work in which all the proteins participate in primary binding, followed by secondary, tertiary and quaternary binding with neighboring proteins without additional import. Thus, 40S hnRNP contains crown-like secondary structure (48 stemloops) and appearance of 6 petal (octamers) rose-like architectures. The proteins are wrapped by RNA. Co-transcriptional folding for RNP fibril of FMR1 gene can produce 2,571 stem-loops with frequency of 1 stem-loop/15.3 nucleotides and 53 40S hnRNP beaded structure. By spliceosome driven reactions, there occurs removal of 16 separate lariated RNPs, joining 17 separate beaded exonic structures and anchoring EJC on each exon junction. Skipping exon 12 has 5'GU, 3'AG and very compact folding pattern with frequency of 1 stem-loop per 12 nucleotides in short exon length (63 nucleotides). 5' end of exon 12 contains SS (Splicing Silencer) element of UAGGU. In exons 10, 15 and 17 where both regular and alternative splice sites exist, SS (hnRNP A1 binding site) is observed at the regular splicing site. End products are mature FMR-1 mRNP, 4 species of Pri-microRNAs derived from introns 7,9,15 and 3'UTR of exon17, respectively. There may also be some other regulatory RNAs containing ALU/Line elements as well.

Isolation, Characterization and Whole-Genome Analysis of Paenibacillus andongensis sp.nov. from Korean Soil

  • Yong Guan;Zhun Li;Yoon-Ho Kang;Mi-Kyung Lee
    • Journal of Microbiology and Biotechnology
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    • v.33 no.6
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    • pp.753-759
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    • 2023
  • The genus Paenibacillus contains a variety of biologically active compounds that have potential applications in a range of fields, including medicine, agriculture, and livestock, playing an important role in the health and economy of society. Our study focused on the bacterium SS4T (KCTC 43402T = GDMCC 1.3498T), which was characterized using a polyphasic taxonomic approach. This strain was analyzed using antiSMASH, BAGEL4, and PRISM to predict the secondary metabolites. Lassopeptide clusters were found using all three analysis methods, with the possibility of secretion. Additionally, PRISM found three biosynthetic gene clusters (BGC) and predicted the structure of the product. Genome analysis indicated that glucoamylase is present in SS4T. 16S rRNA sequence analysis showed that strain SS4T most closely resembled Paenibacillus marchantiophytorum DSM 29850T (98.22%), Paenibacillus nebraskensis JJ-59T (98.19%), and Paenibacillus aceris KCTC 13870T (98.08%). Analysis of the 16S rRNA gene sequences and Type Strain Genome Server (TYGS) analysis revealed that SS4T belongs to the genus Paenibacillus based on the results of the phylogenetic analysis. As a result of the matrix-assisted laser desorption/ionization-time-of-flight mass spectrometry (MALDI-TOF/MS) results, SS4T was determined to belong to the genus Paenibacillus. Comparing P. marchantiophytorum DSM 29850T with average nucleotide identity (ANI 78.97%) and digital DNA-DNA hybridization (dDDH 23%) revealed values that were all less than the threshold for bacterial species differentiation. The results of this study suggest that strain SS4T can be classified as a Paenibacillus andongensis species and is a novel member of the genus Paenibacillus.

Biochemical and Structural Characterization of HP1423 (Y1423_HELPY) from Helicobacter pylori

  • Kim, Ji-Hun;Lee, Ki-Young;Park, Sung-Jean;Lee, Bong-Jin
    • Journal of the Korean Magnetic Resonance Society
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    • v.14 no.1
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    • pp.45-54
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    • 2010
  • HP1423 (Y1423_HELPY) is a conserved hypothetical protein from H. pylori strain 26695. However, Sequence Blast result indicates that HP1423 belongs to S4 (PF01479) superfamily. According to Pfam database, the S4 domain is a small domain consisting of 60-65 amino acid residues, that probably mediates binding to RNA. In this study, we report the sequence-specific backbone resonance assignment of HP1423, which has 84 amino acid residues. We could assign unambiguously about 88% of all $^{1}H_{N}$, $^{15}N$, $^{13}C_{\alpha}$, $^{13}C_{\beta}$ and $^{13}C=O$ resonances. We could not detect the resonances from residues 15-20, and disappearance of these peaks seems to be related with the intermediate-conformational exchange. These assigned NMR peaks of HP1423 can be used for studying the role of protein dynamics in millisecond timescale, and Protein-RNA binding.