• Title/Summary/Keyword: RNA secondary structure

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Continuous Synthesis of Escherichia coli GroEL at a high Temperature

  • Kwak, Young-Hak;Lee, Kyong-Sun;Kim, Ji-Yeon;Lee, Dong-Seok;Kim, Han-Bok
    • Journal of Microbiology
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    • v.38 no.3
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    • pp.145-149
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    • 2000
  • GroEL is a typical molecular chaperone. GroEL synthesis patterns at various culture temperatures in Escherichia coli were investigated in this study. No significant differences in the amount of GroEL produced from the chromosome were found at 30 and 37$^{\circ}C$. However, GroEL production increased 3.4-fold at 42$^{\circ}C$. GroEL synthesis was not transient but continuous at 42$^{\circ}C$, although most heat shock gene expression is known to be transient. To understand the role of the groEL structural gene, a groE promoter-lacZ fusion was constructed. Interestingly , while transcriptional fusion is not thermally inducible, it is inducible by ethanol, suggesting that the secondary structure of the groEL transcript is involved in thermal regulation of the groEL gene. Secondary structures of groE mRNA at 37 and 42$^{\circ}C$ were compared using the computer program RNAdraw. Distinct structures at the two temperatures were found, and these structures may be related to a high level of GroEL expression at 42$^{\circ}C$.

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A highly efficient computational discrimination among Streptococcal species of periodontitis patients using 16S rRNA amplicons

  • Al-Dabbagh, Nebras N.;Hashim, Hayder O.;Al-Shuhaib, Mohammed Baqur S.
    • Korean Journal of Microbiology
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    • v.55 no.1
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    • pp.1-8
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    • 2019
  • Due to the major role played by several species of Streptococcus in the etiology of periodontitis, it is important to assess the pattern of Streptococcus pathogenic pathways within the infected subgingival pockets using a bacterial specific 16S rRNA fragment. From the total of 50 patients with periodontitis included in the study, only 23 Streptococcal isolates were considered for further analyses, in which their 16S rRNA fragments were amplified and sequenced. Then, a comprehensive phylogenetic tree was constructed and in silico prediction was performed for the observed Streptococcal species. The phylogenetic analysis of the subgingival Streptococcal species revealed a high discrimination power of the 16S rRNA fragment to accurately identify three groups of Streptococcus on the species level, including S. salivarius (14 isolates), S. anginosus (5 isolates), and S. gordonii (4 isolates). The employment of state-of-art in silico tools indicated that each Streptococcal species group was characterized with particular transcription factors that bound exclusively with a different 16S rRNA-based secondary structure. In conclusion, the observed data of the present study provided in-depth insights into the mechanism of each Streptococcal species in its pathogenesis, which differ in each observed group, according to the differences in the 16S rRNA secondary structure it takes, and the consequent binding with its corresponding transcription factors. This study paves the way for further interventions of the in silico prediction, with the main conventional in vitro microbiota identification to present an interesting insight in terms of the gene expression pattern and the signaling pathway that each pathogenic species follows in the infected subgingival site.

Effects of Substrate RNA Structure on the Trans-splicing Reaction by Group I Intron of Tetrahymena thermophila (Tetrahymena thermophila의 group I intron에 의한 trans-splicing 반응에 미치는 표적 RNA 구조의 영향분석)

  • 이성욱
    • Korean Journal of Microbiology
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    • v.35 no.3
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    • pp.211-217
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    • 1999
  • Effects of subsh-ate RNA configuration on the tians-splicing reactcon by group I intron ribozyme of Tetralzynzena thern\ulcornerophila were analyzed with substrate RNAs which have been generated to have very stable structures with stem-loop. RNAinapping strategy was perfo~med in vivo as well as in virro to search the mosl accessible siles to the ~irms-splicing ribozymes in the substrate RNAs. Sequences present in the loop of the target RNAs have shown to be well recognized by and reacted with group I inlron ribozymes while sequences present in the stein do not. Thesc results were confirmed with the experiments of trans-cleavage and rmnssplicing reactmn with ihe specific ribozyines recognizing those sequences. Moreover, sequence analysis of the trans-splicing products have shown that irons-splicing reaction can proceed with high fidelity. In conclusion, the secondary structure of substrate RNAs is one of the most important factors to detemine the ribozyme activity.

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Genetic Differentiation among the Mitochondrial ND2 Gene and $tRNA^{Trp}$ Gene Sequences of Genus Rana (Anura) in Korea

  • Lee, Hyuk;Yang, Suh-Yung;Lee, Hei-Yung
    • Animal cells and systems
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    • v.4 no.1
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    • pp.31-37
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    • 2000
  • The genetic variations among six species of Rana from Korea (R. nigro-maculata, R. piancyi, R. dybowskii, R. sp, R. rugosa type A, B and R. amurensis) were investigated using 499 bases of mitochondrial DNA sequences for ND2 (NADH dehydrogenase subunit 2) gene and $tRNA^{Trp}$ gene. Partial sequences of ND2 gene (427 bp) and full sequences of $tRNA^{Trp}$ gene (73 bp) were identified. The level of sequence divergences ranged from 0.2 to 5.2% within species and 4.9-28.0% among 6 species of the genus Rana. The $tRNA^{Trp}$ gene of the genus Rana was composed of 77 nucleotides which showed a two dimensional "cloverleaf" structure. The secondary structure of $tRNA^{Trp}$ was not found compensatory changes which could potentially confound phylogenetic inference. In the neighborjoining tree, brown frogs were clustered first with the level of sequence divergence of 13.20% between R. amurensis and R. dybowskii, and 9% between R. dybowskii and R. sp. supported by 99% bootstrap iterations, respectively. R. nigromaculata and R. plancyi were clustered into another group with 5.1% divergence supported by 100% bootstrap iteration. R. rugosa A 8nd B types were grouped by 4.9% divergence and clustered into the last group with other two groups with 100% bootstrap iterations.

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1H, 15N and 13C resonance assignment and secondary structure prediction of ss-DNA binding protein 12RNP2 precursor, HP0827 from Helicobacter pylori

  • Jang, Sun-Bok;Ma, Chao;Chandan, Pathak Chinar;Kim, Do-Hee;Lee, Bong-Jin
    • Journal of the Korean Magnetic Resonance Society
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    • v.15 no.1
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    • pp.69-79
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    • 2011
  • HP0827 has two RNP motif which is a very common protein domain involved in recognition of a wide range of ssRNA/DNA.We acquired 3D NMR spectra of HP0827 which shows well dispersed and homogeneous signals which allows us to assign 98% of all $^1H_N$, $^{15}N$, $^{13}C_{\alpha}$, $^{13}C_{\beta}$ and $^{13}C$=O resonances and 90% of all sidechain resonances. The sequence-specific backbone resonance assignment of HP0827 can be used to gain deeper insights into the nucleic acids binding specificity of HP0827 in the future study. Here, we report secondary structure prediction of HP0827 derived from NMR data. Additionally, ssRNA/DNA binding assay studies was also conducted. This study might provide a clue for exact function of HP0827 based on structure and sequence.

The Terminal and Internal Hairpin Loops of the ctRNA of Plasmid pJB01 Play Critical Roles in Regulating Copy Number

  • Kim, Sam Woong;Jeong, In Sil;Jeong, Eun Ju;Tak, Je Il;Lee, John Hwa;Eo, Seong Kug;Kang, Ho Young;Bahk, Jeong Dong
    • Molecules and Cells
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    • v.26 no.1
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    • pp.26-33
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    • 2008
  • The plasmid pJB01, a member of the pMV158 family isolated from Enterococcus faecium JC1, contains three open reading frames, copA, repB, and repC. Plasmids included in this family produce counter-transcribed RNA (ctRNA) that contributes to copy number control. The pJB01 ctRNA, a transcript which consists of 54 nucleotides (nts), is encoded on the opposite strand from the copA/repB intergenic region and partially overlaps an atypical ribosome binding site (ARBS) for repB. The ARBS is integrated by the two underlined conserved regions: 5'-TTTTTGTNNNNTAANNNNNNNNNATG-3', and the ctRNA is complementary only to the 5' conserved sequence 5'-TTTTTGT-3'. This complementary sequence is located at a distance from the terminal loop of the ctRNA secondary structure. The ctRNA structure predicted by the mfold program suggests the possible generation of a terminal and an internal hairpin loop. The amount of in vitro translation product of repB mRNA was inversely proportional to the ctRNA concentration. Mutations in the terminal and internal hairpin loops of the ctRNA had inhibitory effects on its binding to the target mRNA. We propose that the intact structures of the terminal and internal hairpin loops, respectively, play important roles in forming the initial kissing and extending complexes between the ctRNA and target mRNA and that these regulate the copy number of this plasmid.

Identification of Nuclear Factors that UV-crosslink to Rev-responsive Element RNA (UV조사에 의해 Rev-responsive element RNA와 결합하는 핵단백질인자의 확인)

  • 박희성;남용석
    • Journal of Life Science
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    • v.7 no.3
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    • pp.161-166
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    • 1997
  • HIV-1 Rev protein plays an important role in regulating the expression of viral structural proteins. It allows the nuclear export and accumulation of unspliced and partially spliced viral mRNA in the cytoplasm. The Rev-responsive element RNA, present in the env gene, forms a higly ordered RNA secondary structure and is required for the Rev-mediated mRNA export. For this process to complete factor(s) are strongly suggested. From our experiments of electrophoretic mobility shift, UV-crosslinking and SDS/PAGE, RRE RNA was found to be recognized to several nuclear factors such as 36/37, 56, 41. 76, 150 kD proteins in the order of reactivity. Among them, 36/37 and 56 kD proteins are more reactive upon a brief UV treatment (5 min) and more persistent in the presence of high amount of nonspecific competitor, heparin. Certain nuclear protein9s) seemed to recognize the RRE RNA structure in competition with Rev to gel mobility shift assay.

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Amazonocrinis thailandica sp. nov. (Nostocales, Cyanobacteria), a novel species of the previously monotypic Amazonocrinis genus from Thailand

  • Tawong, Wittaya;Pongcharoen, Pongsanat;Pongpadung, Piyawat;Ponza, Supat;Saijuntha, Weerachai
    • ALGAE
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    • v.37 no.1
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    • pp.1-14
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    • 2022
  • Cyanobacteria are distributed worldwide, and many new cyanobacterial species are discovered in tropical region. The Nostoc-like genus Amazonocrinis has been separated from the genus Nostoc based on polyphasic methods. However, species diversity within this genus remains poorly understood systematically because only one species (Amazonocrinis nigriterrae) has been described. In this study, two novel strains (NUACC02 and NUACC03) were isolated from moist rice field soil in Thailand. These two strains were characterized using a polyphasic approach, based on morphology, 16S rRNA phylogenetic analysis, internal transcribed spacer secondary structure and ecology. Phylogenetic analyses based on 16S rRNA gene sequences confirmed that the two novel strains formed a monophyletic clade related to the genus Amazonocrinis and were distant from the type species A. nigriterrae. The 16S rRNA gene sequence similarity (<98.1%) between novel strains and all other closely related taxa including the Amazonocrinis members exceeded the cutoff for species delimitation in bacteriology, reinforcing the presence of a new Amazonocrinis species. Furthermore, the novel strains possessed unique phenotypic characteristics such as the presence of the sheath, necridia-like cells, larger cell dimension and akinete cell arrangement in long-chains and the singularity of D1-D1', Box-B, V2, and V3 secondary structures that distinguished them from other Amazonocrinis members. Considering all the results, we described our two strains as Amazonocrinis thailandica sp. nov. in accordance with the International Code of Nomenclature for Algae, Fungi and Plants.

Visualization of RNA secondary structure using vector-based heuristics (벡터에 기반한 휴리스틱을 이용한 RNA 이차 구조의 시각화)

  • 김도형;한경숙
    • Proceedings of the Korean Information Science Society Conference
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    • 1998.10c
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    • pp.633-635
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    • 1998
  • RNA 분자의 이차 구조를 예측하고 예측된 구조를 분석, 평가하기 위하여 시각화하는 작업은 RNA의 구조에 대한 연구에 있어서 가장 필수적인 과정이다. 본 논문은 이차 구조를 시각화하는 알고리즘과 이를 구현한 프로그램을 소개한다. 이 알고리즘은 vector와 vector space를 이용하여 RNA 분자의 구조 요소가 배치될 방향과 공간을 나타낸다. 구조 요소가 겹치지 않도록 배치될 방향과 공간을 효율적으로 찾기 위하여, 구조 요소를 배치하는 순서에 관한 휴리스틱과 구조 요소를 배치하는 방법에 관한 휴리스틱을 사용한다. 기존의 시각화 알고리즘이 구조요소를 순차적으로 배치하면서 겹침 현상이 발생하면, 이를 제거 하기 위하여 이미 배치한 구조 요소들을 재배치 하거나 변형하는 것에 반하여, 이 알고리즘은 사용자의 판단이나 수작업에 의존하지 않고 겹침 현상이 거의 없는 이차 구조를 효율적으로 생성한다는 점에서 기존 방법을 많이 개선하였다.

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Visualization and EDiting of RNA Secondary Structure (RNA 이차 구조의 시각화와 편집)

  • Han, Gyeong-Suk;Kim, Do-Hyeong
    • Journal of KIISE:Computer Systems and Theory
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    • v.26 no.5
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    • pp.539-548
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    • 1999
  • RNA 분자의 이차 구조를 예측하고 예측된 구조를 분석 평가하기 위하여 시각화하는 작업은 RNA의 구조에 대한 연구에 있어서 가장 필수적인 과정이다. 본 논문은 이차 구조를 시각화하고 편집하는 실용적이면서 새로운 알고리즘을 소개한다. 이 시각화 알고리즘은 벡터와 백터공간을 이용하여 RNA 분자의 구조 요소가 배치될 방향과 영역을 나타낸다. 구조 요소가 겹치지 않도록 배치될 방향ㄹ과 공간을 효율적으로 찾기위하여 구조 요소를 배치하는 순서에 관한 휴리스틱과 구조 요소를 배치하는 방법에 관한 휴리스틱을 사용한다. 이 시각화 알고리즘은 IBM PC/Windows 95 환경에서VizQFolder 라는 프로그램으로 구현되었다. 실험 결과는 VizQFolder 가 이미 배치한 구조 요소들을 재배치하거나 변형하지 않으면서 helix의 회전만으로 겹침 현상이 거의 없는 이차구조를 효율적으로 생성한다는 것을 보인다.