• Title/Summary/Keyword: RNA secondary structure

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Nucleotide Sequence and Secondary Structure of 16S rRNA from Sphingomonas chungbukensis DJ77 (Sphingomonas chungbukensis DJ77의 16S rRNA 염기서열과 이차구조)

  • Lee Kwan-Young;Kwon Hae-Ryong;Lee Won-Ho;Kim Young-Chang
    • Korean Journal of Microbiology
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    • v.41 no.2
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    • pp.125-128
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    • 2005
  • A 16S ribosomal RNA gene from S. chungbukensis DJ77 has been sequenced. This sequence had a length of 1,502 bp and was extended for 29 bp at 5' and for 37 bp at 3' from the partial sequence (1,435 bp) registered in 2000 year. Besides, 1 bp was newly added near to the 3' end. We made the secondary structure of the 16S rRNA based on E. coli model and found four specific regions. We found constant and variable regions in genus Sphingomonas as the result of multiple alignment of 16S rRNA gene sequences from Sphingomonas spp. and S. chungbukensis DJ77. We found a stem loop structure in S. chungbukensis DJ77, which was only discovered in C. jejuni to date. It showed the structural agreement despite the difference of the sequences from the both organisms. Finally, S. chungbukensis DJ77 belonged to cluster II (Sphingobium) group, after the classification using phylogenetic analysis and nucleotide signature analysis.

SECONDARY STRUCTURE OF THE PANHANDLE RNA OF INFLUENZA VIRUS A STUDIED BY NMR SPECTROSCOPY

  • Cheong, Hae-Kap;Park, Byong-Seok;Chaejoon Cheong
    • Proceedings of the Korean Biophysical Society Conference
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    • 1996.07a
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    • pp.31-31
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    • 1996
  • The double-stranded panhandle structure of the influenza virus RNA is important for the replication, transcription and packaging into the virion of the vRNA. The solution structure of a 34-nucleotide-long RNA which contains the conserved panhandle sequences has been investigated by one- and two-dimensional NMR spectroscopies. (omitted)

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Determination of the Structure of 5S rRNA from Xanthomonas citri (Xanthomonas citri의 5S rRNA 의 구조 결정)

  • Bongrae Cho;Myung-Un Choi;Se Won Seh;ahei Ihm;Moonjoo Koh;Inwon Park
    • Journal of the Korean Chemical Society
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    • v.36 no.3
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    • pp.460-465
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    • 1992
  • The structure of th 5S rRNA isolated from Xanthomonas citri was determined by enzymatic and chemical degradation methods. It consists of 119 nucleotides and contains no modified nucleosides. As does the 5S rRNA of X. maltophilia, it contains an additional uridine residue on the 5'-terminus. The secondary structure of the 5S rRNA from X. citri was almost identical to the generalized models proposed by many other workers [De Wachter et al., Biochimie, 64, 311 (1982); Specht et al., Nucleic Acids Res., 18, 2215 (1990); Cho et al., Proceedings of the First Symposium on Biomolecules, p. 9 (1991)]. The secondary structure of 5S rRNA from X. citri consists of five helices, five loops and two bulges. This 5S rRNA has a uridine residue at position 66 as a bulge.

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Proper NMR methods for studying RNA thermometers

  • Kim, Won-Je;Kim, Nak-Kyoon
    • Journal of the Korean Magnetic Resonance Society
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    • v.19 no.3
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    • pp.143-148
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    • 2015
  • In some pathogenic bacteria, there are RNA thermometers, which regulate the production of virulence associated factors or heat shock proteins depending on temperature changes. Like a riboswitches, RNA thermometers are located in the 5'-untranslated region and involved translational gene regulatory mechanism. RNA thermometers block the ribosome-binding site and start codon area under the $37^{\circ}C$ within their secondary structure. After bacterial infection, increased the temperature in the host causes conformations changes of RNA, and the ribosome-binding site is exposed for translational initiation. Because structural differences between open and closed forms of RNA thermometers are mainly mediated by base pairing changes, NMR spectroscopy is a very useful method to study these thermodynamically changing RNA structure. In this review, we briefly provide a fundamental function of RNA thermometers, and also suggest a proper NMR experiments for studying RNA thermometers.

Backbone assignment of the anticodon binding domain of human Glycyl-tRNA synthetase

  • Mushtaq, Ameeq Ul;Cho, Hye Young;Byun, Youngjoo;Jeon, Young Ho
    • Journal of the Korean Magnetic Resonance Society
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    • v.20 no.2
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    • pp.50-55
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    • 2016
  • Backbone $^1H$, $^{13}C$ and $^{15}N$ resonance assignments are presented for the anticodon binding domain (residues 557-674) of human glycyl-tRNA synthetase (GRS). Role of the anticodon binding domain (ABD) of GRS as an anticancer ligand has recently been reported and its role in other diseases like Charcot-Marie-Tooth (CMT) and polymyositis have increased its interest. NMR assignments were completed using the isotope [$^{13}C/^{15}N$]-enriched protein and chemical shifts based secondary structure analysis with TALOS+ demonstrate similar secondary structure as reported in X-ray structure PDB 2ZT8, except some C-terminal residues. NMR signals from the N-terminal residues 557 to 571 and 590 to 614 showed very weak or no signals exhibiting dynamics or conformational exchange in NMR timescale.

Effect of random Shine-Dalarno sequence on the expression of Bovine Growth Hormone Gene in Escherichia coli (대장균에서 무작위 샤인-달가노 서열이 소성장호르몬 유전자 발현에 미치는 영향)

  • 나경수;나경수;백형석;이용세
    • Journal of Life Science
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    • v.10 no.4
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    • pp.422-430
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    • 2000
  • In order to search for the effects of Shine-Dalgarno (SD) sequence and nucleotide sequence of spacer region (SD-ATG) on bGH expression, oligonucleotides containing random SD sequences and a spacer region were chemically synthesized. The distance between SD region and initiation codon (ATG) was fixed to 9 nucleotides in length. The expression vectors have been constructed using pT7-1 vector containing a T7 promoter. Positive clones were screened with colony hybridization and named pT7A or pT7B plasmid series. The selected clones were confirmed by DNA sequencing and finally, 19 clones having various SD combinations were obtained. When bovine growth hormone was induced by IPTG in E. coli BL21(DE3), all cells harboring these plasmids produced a detectable level of bGH in western blot analysis. However, various SD sequences did not affect on bGH expression, indicating that the sequences of SD and the spacer region did not sufficiently destabilize mRNA secondary structure of bGH gene. Therefore, these results indicate that the disruption of mRNA secondary structure might be a major factor for regulating bGH expression in the translational initiation process.

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Designing Signal Peptides for Efficient Periplasmic Expression of Human Growth Hormone in Escherichia coli

  • Jeiranikhameneh, Meisam;Moshiri, Farzaneh;Falasafi, Soheil Keyhan;Zomorodipour, Alireza
    • Journal of Microbiology and Biotechnology
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    • v.27 no.11
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    • pp.1999-2009
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    • 2017
  • The secretion efficiency of a protein in a Sec-type secretion system is mainly determined by an N-terminal signal peptide and its combination with its cognate protein. Five signal peptides, namely, two synthetic Sec-type and three Bacillus licheniformis alpha-amylase-derived signal peptides, were compared for periplasmic expression of the human growth hormone (hGH) in E. coli. Based on in silico predictions on the signal peptides' cleavage efficiencies and their corresponding mRNA secondary structures, a number of amino acid substitutions and silent mutations were considered in the modified signal sequences. The two synthetic signal peptides, specifically designed for hGH secretion in E. coli, differ in their N-terminal positively charged residues and hydrophobic region lengths. According to the mRNA secondary structure predictions, combinations of the protein and each of the five signal sequences could lead to different outcomes, especially when accessibility of the initiator ATG and ribosome binding sites were considered. In the experimental stage, the two synthetic signal peptides displayed complete processing and resulted in efficient secretion of the mature hGH in periplasmic regions, as was demonstrated by protein analysis. The three alpha-amylase-derived signal peptides, however, were processed partially from their precursors. Therefore, to achieve efficient secretion of a protein in a heterologous system, designing a specific signal peptide by using a combined approach of optimizations of the mRNA secondary structure and the signal peptide H-domain and cleavage site is recommended.

Nucleotide sequence analysis of the 5S ribosomal RNA gene of the mushroom tricholoma matsutake

  • Hwang, Seon-Kap;Kim, Jong-Guk
    • Journal of Microbiology
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    • v.33 no.2
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    • pp.136-141
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    • 1995
  • From a cluster of structural rRNA genes which has previsouly been cloned (Hwang and Kim, in submission; J. Microbiol. Biotechnol.), a 1.0-kb Eco RI fragment of DNA which shows significant homology to the 25S and rRNA s of Tricholoma matsutake was used for sequence analysis. Nucleotide sequence was bidirectionally determined using delection series of the DNA fragment. Comparing the resultant 1016-base sequence with sequences in the database, both the 3'end of 25S-rRNA gene and 5S rRNA gene were searched. The 5S rRNA gene is 118-bp in length and is located 158-bp downstream of 3'end of the 25S rRNA gene. IGSI and IGS2 (partial) sequences are also contained in the fragment. Multiple alignment of the 5S rRNA sequences was carried out with 5S rRNA sequences from some members of the subdivision Basidiomycotina obtained from the database. Polygenetic analysis with distance matrix established by Kimura's 2-parameter method and phylogenetic tree by UPGMA method proposed that T. matsutake is closely related to efibulobasidium allbescens. Secondary structure of 5S rRNA was also hypothesized to show similar topology with its generally accepted eukaryotic counterpart.

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