• Title/Summary/Keyword: RNA4

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Some RNases Involved in the Processing of Bacteriophage T4 RNA (박테리오파지 T4 tRNA의 프로세싱에 관여하는 몇가지 RNase들)

  • Thong-Sung Ko
    • Journal of the Korean Chemical Society
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    • v.26 no.6
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    • pp.396-402
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    • 1982
  • Bacteriophage T4 tRNA processing in E. coli mutant strains defective in RNase Ⅲ, RNase E$^-$, and RNase P, respectively, singly or in combinations, was investigated. In $RNase E^- strains, a RNA band, which would be referred as 9S RNA, accumulates, while in RNase$ P^-$ strains, lower band of 6S double band is accumulated. In RNase III$^-$ strains, the production of tRAN$^{Gln}$ coded by T4 tRNA gene cluster, is severely depressed and also production of species 1 RNA, which is coded by T4 DNA but not by the tRNA gene cluster, is in somewhat depressed amounts; on the other hand, at the same time, an upper band of 6S double bands, coded by T4 tRNA gene cluster, is accumulated in rather greater amounts as compared to the RNase $^+$ strain. The upper band RNA of the 6S double band, however, does not appear to be a precursor to the tRNA$^{Gln}$. The present work points to the lack of evidence for an essential cleavage role of RNase Ⅲ, although there must be a role for the RNase Ⅲ in the T4 tRNA processing.

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Molecular cDNA cloning and unusual RNA editings of NAD4 gene from Zea mays mitochondrion (옥수수 미토콘드리아 NAD4유전자의 cDNA cloning과 특이한 RNA editing 현상)

  • 설일환
    • Journal of Life Science
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    • v.8 no.2
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    • pp.203-207
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    • 1998
  • NAD4 as a gene encoding NADH dehydrogenase subunit 4 in the micodhondrion from maize has been cloned using RT-PCR and sequenced for examining RNA edited sites. Analysis of mt cDNA sequences showed the typical RNA editing patterns and unusual base changes as well;RNA editing from cDNA sequences occured base change from c to U in most cases, however transitions from t to g and G to A were also observed. Even though those editings appared to be occurred randomly, RNA edited sites showed mostly in exon 1 and exon 4 regions, when compared with NAD4 cDNA from wheat, locations of edited sites did not consistent with each other suggesting that the phenomenon of RNA editing occured randomly not site-specific manner.

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Inhibition of HIV-1 Replication in CD4+ Peripheral Blood Lymphocytes by Intracellular Expression of RNA Aptamer (RNA aptamer 발현을 통한 CD4+ peripheral blood lymphocytes에서의 인간 면역결핍 바이러스의 증식 억제)

  • Lee, Seong-Uk
    • Korean Journal of Microbiology
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    • v.39 no.4
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    • pp.235-241
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    • 2003
  • We have previously demonstrated that intracellular expression of an RNA aptamer termed RRE40, which was selected in vitro to bind HIV Rev 10-fold much tighter than wild-type RRE, efficiently protected human CD4+ T cell line, CEM, from HIV-1. In this study, to evaluate the efficacy of the RRE40 RNA in clinical settings, polyclonal CD4+ peripheral blood lymphocytes (PBLs) were transduced with retroviral vectors expressing RRE40 decoy RNA and then challenged with clinical isolates of HIV-1. In contrast to the control cells transduced with vectors expressing control tRNA, intracellular expression of RRE40 RNA more effectively inhibited HIV-1 replication in CD4+ PBLs. However, transient and diminished inhibition, rather than complete inhibition, of HIV-1 replication in PBLs expressing RRE40 decoys have been observed. These results suggest that RRE40 decoy RNA would be useful to inhibit HIV-1 replication in cells. However, development of more efficient gene transfer protocols and/or more effective decoy RNAs would be needed to apply RNA decoy to modulate HIV-1 patient.

Complete Genome Sequence of the RNAs 3 and 4 Segments of Rice stripe virus Isolates in Korea and their Phylogenetic Relationships with Japan and China Isolates

  • Jonson, Miranda Gilda;Choi, Hong-Soo;Kim, Jeong-Soo;Choi, Il-Ryong;Kim, Kook-Hyung
    • The Plant Pathology Journal
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    • v.25 no.2
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    • pp.142-150
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    • 2009
  • The complete genome sequences of RNA3 and RNA4 of the 13 different Rice stripe virus (RSV) isolates were determined and characterized in this study to address the possible causes of the recent re-emergence of RSV that affected many rice fields in Korea. The genome size of each RNA segment varied among isolates and significant differences were observed in the intergenic region. There was up to 4% average divergence in the RNA4 nucleotide sequence among 13 Korean isolates and only 1.4% in the RNA3. Phylogenetic relationships among different Korean isolates revealed that there were at least 2 types of RNA3 and 4 distinct types of RNA4 genomes present in Korea. However, Korean isolates with one type of RNA3 predominate over the other while the occurrences of the RSV Korean isolates with the 4 types of RNA4 genome were not correlated to specific geographical areas. Results further indicate that RNA4 had diverged more than RNA3 and these differences in accumulation of mutations in the individual RNA segments indicate that genetic reassortment were likely to contribute to the genetic divergence in the 13 Korean isolates. All of the Korean-RNA3 sequences except for one isolate grouped with Chinese isolates (JY and Z). In contrast, the RNA 4 sequences segregated together with either Chinese (JY and Z) and Japanese (M and T) isolates but genetic relationships of Korean isolates- RNAs 3 and 4 segments to Chinese-Y isolate were low. Altogether, these results suggest that the occurrence of mixtures of RNAs 3 and 4 genotypes in the natural population of RSV may have contributed to the sudden outbreak in Korea.

RNA content of Bombyx mori egg during its development and irradiation effect on its RNA content (Part. 1) (가잠란 발육과정에 따른 RNA 함량의 변동 및 방사선이 잠란 RNA 함량에 미치는 영향 (1))

  • 김영수;이기영;최병희
    • Journal of Sericultural and Entomological Science
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    • v.3
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    • pp.23-28
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    • 1963
  • The RNA content of fertilized egg of Bombyx mori was shown a continuing great increase reaching peak at the 5th day after egg laying and a slight decrease there after. Such a change of RNA content is considered to be directly associated with the formation and development of egg embryo of silk worm. (2) The RNA content of nonfertilized egg is much less than that of fertilized one at first day of egg laying and it increased slightly until 4 th day after egg laying then decreased. (3) The RNA content of fertilized egg irradiated by gamma-ray (3,000 r) was shown a slight increase until 2nd day after irradiation, but no change was observed there after. This fact shows that irradiation suppressed the biosynthesis of RNA silk worm egg. (4) The RNA content of HCl treated silk worm egg was shown a continuing steep rise until 7 th day after the acid treat, while no change was observed in the non-treated egg. The RNA content of HCl treated egg with irradiation of gamma-ray (1,500 r), decreased until 3 rd day after irradiation in contrast to that of non-irradiated group, but it increased rapidly from 4 th day until 7 th day after acid treating.

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Effect of hnRNP-like protein THO4 on growth and mRNA export in fission yeast (분열효모에서 hnRNP-유사 단백질인 THO4가 생장 및 mRNA 방출에 미치는 영향)

  • Park, Jin Hee;Lee, Sojeong;Yoon, Jin Ho
    • Korean Journal of Microbiology
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    • v.54 no.2
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    • pp.91-97
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    • 2018
  • The evolutionally conserved TREX complex member, Yra1/ALY, belongs to the REF (RNA and export factor binding proteins) family of hnRNP-like proteins, which has been implicated in multiple processes including transcription, nuclear RNA stability, and mRNA export. Fission yeast, Schizosaccharomyces pombe, genome encodes two members of REF proteins. In addition to Mlo3 known previously as an mRNA export factor, there is the other REF protein, Tho4, which is predicted as a component of THO complex. Here we showed that deletion of tho4 (SPBC106.12c) gene does not inhibit both growth and nuclear mRNA export. However, overexpression of tho4 displays growth retardation and slight accumulation of $poly(A)^+$ RNA in the nucleus. Neither ${\Delta}tho4$ ${\Delta}mlo3$ nor ${\Delta}tho4$ ${\Delta}mex67$ double mutants exhibit additive growth defect. Moreover, yeast two-hybrid and co-immunoprecipitation analysis did not show that the Tho4 protein interacted with any members of TREX complex and mRNA export factor Rae1. Contrary to expectation, these observations support that the S. pombe Tho4 is not a component of TREX complex, and not directly involved in bulk mRNA export from the nucleus.

Enhanced Delivery of siRNA Complexes by Sonoporation in Transgenic Rice Cell Suspension Cultures

  • Cheon, Su-Hwan;Lee, Kyoung-Hoon;Kwon, Jun-Young;Choi, Sung-Hun;Song, Mi-Na;Kim, Dong-II
    • Journal of Microbiology and Biotechnology
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    • v.19 no.8
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    • pp.781-786
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    • 2009
  • Small interfering synthetic double-stranded RNA (siRNA) was applied to suppress the expression of the human cytotoxic-T-Iymphocyte antigen 4-immunoglobulin (hCTLA4Ig) gene transformed in transgenic rice cell cultures. The sequence of the 21-nucleotide siRNA was deliberately designed and synthesized with overhangs to inactivate the expression of hCTLA4Ig. The chemically synthesized siRNA duplex was combined with polyethyleneimine (PEl) at a mass ratio of 1:10 (0.33 ${\mu}g$ siRNA:3.3 ${\mu}g$ PEl) to produce complexes. The siRNA complexes (siRNA+PEI) were labeled with Cy3 in order to subsequently confirm the delivery by fluorescent microscopy. In addition, the cells were treated with sonoporation at 40 kHz and 419W for 90 s to improve the delivery. The siRNA complexes alone inhibited the expression of hCTLA4Ig to 45% compared with control. The siRNA complexes delivered with sonoporation downregulated the production of hCTLA4Ig to 73%. Therefore, we concluded that the delivery of siRNA complexes into plant cells could be enhanced successfully by sonoporation.

Determination of Nucleotide Sequences of cDNA from Cucumber Mosaic Virus-As RNA4 (As계의 오이 모자이크 바이러스 RNA4의 염기서열 결정)

  • 김상현;박원목;이세영;박영인
    • Korean Journal Plant Pathology
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    • v.12 no.2
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    • pp.176-181
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    • 1996
  • Aster yomena로부터 분리한 오이 모자이크 바이러스(cucumber mosaic virus) (CMV-As)의 RNA4로부터 완전한 길이의 cDNA를 합성하고 그 전체적인 염기서열(1,043 nt`s)을 결정하였다. CMV-As RNA4는 73개의 염기로 구성된 5`말단의 leader 부위, 657개의 염기로 구성된 외피단백질(coat protein) 유전자 부위 및 312개의 염기로 구성된 3` 말단의 비번역 부위로 구성되어 있음을 확인하였다. 외피단백질 유전자 부위의 염기서열을 다른 계통의 CMV와 비교해 볼 때 그 염기서열이 보전적으로 존재하고 있으나 그 외의 부분은 다양함을 확인하였다. 특히 3` 말단부위의 61개의 염기로 구성된 부위(959-1019)는 다른 계통의 CMV에서는 상당히 유사하지만 CMV-As도 다른 CMV처럼 tRNA와 유사한 구조를 역시 형성함을 확인하였다. CMV-As의 RNA4 염기서열을 다른 계통의 CMV와 비교할 때 CMV-I17F와 가장 유사하였으며(91.9%) S형의 CMV-M과는 가장 낮은 동일성을 보였다(71.1%). 외와 같은 염기성열의 비교 결과와 EcoRI 제한효소 인식부위의 존재로 미루어 CMV-As는 WT형으로 분류된다.

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Oxidative stress causes Alu RNA accumulation via PIWIL4 sequestration into stress granules

  • Hwang, Yeo Eun;Baek, Yu Mi;Baek, Ahruem;Kim, Dong-Eun
    • BMB Reports
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    • v.52 no.3
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    • pp.196-201
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    • 2019
  • The Alu element, the most abundant transposable element, is transcribed to Alu RNA. We hypothesized that the PIWI protein regulates the expression of Alu RNA in retinal pigment epithelial (RPE) cells, where accumulated Alu RNA leads to macular degeneration. Alu transcription was induced in RPE cells treated with $H_2O_2$. At an early stage of oxidative stress, PIWIL4 was translocated into the nucleus; however, subsequently it was sequestered into cytoplasmic stress granules, resulting in the accumulation of Alu RNA. An elevated amount of Alu RNA was positively correlated with the disruption of the epithelial features of RPE via induction of mesenchymal transition. Therefore, we suggest that oxidative stress causes Alu RNA accumulation via PIWIL4 sequestration into the cytoplasmic stress granules.

Post-transcriptional and translational regulation of mRNA-like long non-coding RNAs by microRNAs in early developmental stages of zebrafish embryos

  • Lee, Kyung-Tae;Nam, Jin-Wu
    • BMB Reports
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    • v.50 no.4
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    • pp.226-231
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    • 2017
  • At the post-transcriptional and translational levels, microRNA (miRNA) represses protein-coding genes via seed pairing to the 3' untranslated regions (UTRs) of mRNA. Although working models of miRNA-mediated gene silencing are successfully established using miRNA transfections and knockouts, the regulatory interaction between miRNA and long non-coding RNA (lncRNA) remain unknown. In particular, how the mRNA-resembling lncRNAs with 5' cap, 3' poly(A)-tail, or coding features, are regulated by miRNA is yet to be examined. We therefore investigated the functional interaction between miRNAs and lncRNAs with/without those features, in miRNA-transfected early zebrafish embryos. We observed that the greatest determinants of the miRNA-mediated silencing of lncRNAs were the 5' cap and 3' poly(A)-tails in lncRNAs, at both the post-transcriptional and translational levels. The lncRNAs confirmed to contain 5' cap, 3' poly(A)-tail, and the canonical miRNA target sites, were observed to be repressed in the level of both RNA and ribosome-protected fragment, while those with the miRNA target sites and without 5' cap and 3' poly(A)-tail, were not robustly repressed by miRNA introduction, thus suggesting a role as a miRNA-decoy.