• Title/Summary/Keyword: 3'-rRNase

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Species-Specific Cleavage by RNase E-Like Enzymes in 5S rRNA Maturation

  • RYOU SANG-MI;KIM JONG-MYUNG;YEOM JI-HYUN;KIM HYUN-LI;GO HA-YOUNG;SHIN EUN-KYOUNG;LEE KANGSEOK
    • Journal of Microbiology and Biotechnology
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    • v.15 no.5
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    • pp.1100-1105
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    • 2005
  • Previous work has identified a Streptomyces coelicolor gene, rns, encoding a 140 kDa protein (RNase ES) that exhibits the endoribonucleolytic cleavage specificity characteristic of RNase E and confers viability on and allows the propagation of E. coli cells lacking RNase E. Here, we identify a putative S. coelicolor 9S rRNA sequence and sites cleaved by RNase ES. The cleavage of the S. coelicolor 9S rRNA transcript by RNase ES resulted in a 5S rRNA precursor (p5S) that had four and two additional nucleotides at the 5' end and 3' ends of the mature 5S rRNA, respectively. However, despite the similarities between RNase E and RNase ES, these enzymes could accurately process 9S rRNA from just their own bacteria, indicating that these ancient enzymes and the rRNA segments that they attack appear to have co-evolved.

Induction of Ribosomal Ribonuclease during Catabolic Repression in Saccharomyces uvarum (Saccharomyces uvarum의 Catabolic Repression 시기에 유도되는 Ribosomal Ribonuclease에 대한 연구)

  • Yoon, Seong-Nyo;Lee, Ki-Sung;Choi, Yong-Keel
    • The Korean Journal of Mycology
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    • v.14 no.3
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    • pp.201-207
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    • 1986
  • In order to study subcellular locality and characteristics of ribonuclease in Saccharomyces uvarum, subcelllar fractions $45,000{\times}g$ pellet fraction, post ribosomal fraction and ribosome fraction were extracted during late log, stationary phase and sugar starvation conditions. Ribonuclease activity was significantly increased in ribosomal fraction under stationary and sugar starvation conditions. Ribosomal ribonuclease was extracted by EDTA plus streptomycin sulfate and ammonium sulfate precipitation. The amount of ribosome in stationary and sugar starvation condition was decreased three to six fold as compared to that in the early log phase. The end products of ribosomal ribonuclease were detected by thin layer chromatography. It is postulated that the increase of ribosomal ribonuclease activity under sugar starvation results from 5'-rRNase, while the increase of rRNase activity under stationary phase results from 3'-rRNase.

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The Effect of Gibbrellic Acid and Abscisic Acid on Ribonucleic Acid and Ribonuclease in Barley Coleoptiles (Gibbrellic Acid와 Abscisic Acid가 보리 초엽(?葉)의 핵산(核酸) 및 핵산분해효소(核酸分解酵素)에 미치는 영향(影響))

  • Suh, Yong-Taek
    • Applied Biological Chemistry
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    • v.20 no.2
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    • pp.242-246
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    • 1977
  • In the barly coleptile sections treated with either $1{\times}10^{-5}M$ abscisic acid (ABA) or $1{\times}10^{-5}M$ gibberellic acid (GA), the time course changes of ribonuclease (RNase) activity and ribonucleic acid (RNA) profiles were studied. The results may be summarized as follows: 1. While GA suppressed RNase activity, ABA activated it. 2. High level of s-RNA and low level of r-RNA compared with normal plant sections in hormone-untreated coleoptiles seemed to be the results of increased RNase activity in the incubation period. 3. While GA retarded the decomposition of r-RNA, ABA activated it and the results seemed to be related with RNase activity. 4. GA activated the synthesis of RNA-DNA component, and ABA suppressed it. 5. Increase in the amount of s-RNA with the treatment of ABA may be due to the decomposition of r-RNA.

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Whole-mount in situ Hybridization of Mitochondrial rRNA and RNase MRP RNA in Xenopus laevis Oocytes

  • Jeong, Sun-Joo
    • Animal cells and systems
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    • v.2 no.4
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    • pp.529-538
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    • 1998
  • In order to analyze the intracellu1ar localization of specific RNA components of ribonucleoproteins (RNP) in Xenopus oocytes, a modified protocol of whole-mount in situ Hybridization is presented in this paper, Mitochondria specific 12S rRNA probe was used to detect the amplification and distribution of mitochondria in various stages of the oocyte life cycle, and the results were found to be consistent with previously known distribution of mitochondria. The results with other specific probes (U1 and U3 small nuclear RNAs, and 5S RNA) also indicate that this procedure is generally effective in localizing RNAs in RNP complexes even inside organelles. In addition, the RNA component of RNase MRP, the RNP with endoribo-nuclease activity, localize to the nucleus in various stages of the oocyte life cycle. Some of MRP RNA, however, were found to be localized to the special population of mitochondria near the nucleus, especially in the active stage of mitochondrial amplification. It suggests dual localization of RNase MRP in the nucleus and mitochondria, which is consistent with the proposed roles of RNase MRP in mitochondrial DNA replication and in rRNA processing in the nucleolus.

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PURIFICATION AND PROPERTIES OF EXTRACELLULAR NUCLEASE(S) FROM RUMEN CONTENTS OF BUBALUS BUBALIS

  • Sinha, P.R.;Dutta, S.M.
    • Asian-Australasian Journal of Animal Sciences
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    • v.3 no.2
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    • pp.115-120
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    • 1990
  • Extracellular nuclease(s) in buffalo rumen fluid were purified from strained rumen fluid by a procedure involving Seitz filtration, acetone fractionation and gel filtration on Sephadex G-100. The enzyme resolved into two peaks exhibiting both DNase and RNase activities. The molecular weight of enzyme corresponding to peaks I and II were approximately 30,000 and 12,000 respectively. The properties of enzymes from the two peaks, however, were same. Optimum temperature for both DNase and RNase activities was at $50^{\circ}C$. Whereas DNase activity was stable upto $60^{\circ}C$, RNase activity was stable only up to $50^{\circ}C$. DNase activity recorded two pH optima, one at pH 5.5 and the other at pH 7.0. RNase activity recorded a broad pH optimum between pH 6.0-8.0. pH stability of the enzyme coincided with pH optima for both the activities. DNase activity was stimulated by $Mg^{2+}$ and $Mn^{2+}$ and inhibited by $Fe^{2+}$, $Zn^{2+}$, $Hg^{2+}$ and $Ag^+$. RNase activity was also stimulated by $Mg^{2+}$ and $Mn^{2+}$ and inhibited by $Cu^{2+}$, $Fe^{2+}$, $Zn^{2+}$, $Hg^{2+}$ and $Ag^+$. Reducing agents stimulated both the activities.

Variation of RNase activities and nucleic acid content of non-irradiated and irradiated eggs of Bombyx mori during early development of embryo (잠란(蠶卵)의 초기발육과정(初期發育過程)에 따르는 RNase활성(活性) 및 핵산량(核酸量)의 변동(變動) 및 그 X선조사(線照射)에 의한 영향(影響)에 관(關)한 연구(硏究))

  • Lee, K.Y.;Cheon, H.W.
    • Applied Biological Chemistry
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    • v.15 no.2
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    • pp.163-168
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    • 1972
  • Previously identified female pupae were X-irradiated with a dose of 1000r one day prior to moth transformation. Female mothes from irradiated and non-irradiated pupae were copulated with normal male ones and allowed to lay eggs. Fertilized eggs were collected at 6 intervals such as 5, 15, 45, 90 minutes, 12 and 40 hours after laying, and deep-freezed immediately after each collection until measurements. RNase activity and nucleic acid content were determined with each sample and following results were obtained. 1) It was proved to exist two RNases in silk worm eggs as in mammalian tissues, one active maximally at pH 5.8 and the other at pH 8.0, and the acid RNase activity was much higher than that of alkaline RNase. 2) The activity of acid and alkaline RNases increased remarkably during early development of the embryo of silk worm eggs, reaching the maximum activity at 45 minutes from laying time in non-irradiated group. There was no appreciable difference in two RNase activities for 45 minutes after laying in both control and irradiated groups, but the activity of acid and alkaline RNases in latter group was three times as much as that in former group, at 90 minutes from laying time and it was also found the acid RNase activity was 1.8 times higher than alkaline one in irradiated group. 3) The RNA-P content of control group increased considerably for initial 45 minutes, followed by a decline 45 minutes later with sight but steady increase thereafter. The RNA-P content of irradiated group, however, increased at initial 5 minutes, followed by a marked fall 90 minutes after laying, with no change thereafter. The DNA-P of control group showed a sharp increase for initial 45 minutes, followed by a decline 45 minutes later with no appreciable change thereafter, whereas that of irradiated group showed an increase at initial 15 minutes, followed by a sharp decline for following 45 minutes with a gradual increase thereafter. It was thus proved that the synthesis of nucleic acid in silk worm eggs was much suppressed by X-irradiation during early development of embryo. 4) The RNase activity varied in parallel with the RNA-P content in control group, but the RNA-P content in irradiated group was shown to be minimum value in concidence with the maximum activity of both RNases.

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Identification of a novel circularized transcript of the AML1 gene

  • Xu, Ai-Ning;Chen, Xiu-Hua;Tan, Yan-Hong;Qi, Xi-Ling;Xu, Zhi-Fang;Zhang, Lin-Lin;Ren, Fang-Gang;Bian, Si-Cheng;Chen, Yi;Wang, Hong-Wei
    • BMB Reports
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    • v.46 no.3
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    • pp.163-168
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    • 2013
  • The AML1 gene is an essential transcription factor regulating the differentiation of hematopoietic stem cells into mature blood cells. Though at least 12 different alternatively spliced AML1 mRNAs are generated, three splice variants (AML1a, AML1b and AML1c) have been characterized. Here, using the reverse transcription-polymerase chain reaction with outward-facing primers, we identified a novel non-polyadenylated transcript from the AML1 gene, with exons 5 and 6 scrambled. The novel transcript resisted RNase R digestion, indicating it is a circular RNA structure that may originate from products of mRNA alternative splicing. The expression of the novel transcript in different cells or cell lines of human and a number of other species matched those of the canonical transcripts. The discovery provides additional evidence that circular RNA could stably exist in vivo in human, and may also help to understand the mechanism of the regulation of the AML1 gene transcription.

Molecular Analysis of double-stranded RNA in Abnormal Growing Oyster-Mushrooms, Pleurotos florida and P. ostreatus due to Virus Infection (Virus 이병(罹病) 느타리버섯 (Pleurotus)으로부터 double-stranded RNA 의 분리(分離))

  • Go, Seung-Joo;Park, Yong-Hwan;Shin, Gwan-Chull;Wessels, Josep G.H.
    • The Korean Journal of Mycology
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    • v.20 no.3
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    • pp.234-239
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    • 1992
  • The experiment was performed to find out the possibilities to detect virus infection in oyster mushrooms, Pleurotus species by analysis of doublestranded ribonucleic acid (ds RNA). Ds RNA segments were extracted from virus infected isolates which grew abnormally. But virus free isolates didn't show any ds RNA segments. The ds RNA was consisted of one large segment of 8100 base pairs (bp) and 4 smaller segments with 2170, 2120, 1980 and 1984 bp. Whereas, cell free virus particles showed only one larger ds RNA segment. The ds RNA was dissolved by RNase A in low salt, 0.1 M SSC and melted at $85^{\circ}C$. It was possible to use the ds RNA analysis for detecting virus infection directly from the host cells.

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The Effect of Gibberellic and Abscisic Acids on The Synthesis of Ribonucleic Acid in Seeds and Coleoptiles of Barley (Giberellic acid와 Abscisic acid가 대맥종자(大麥種子) 및 초엽(?葉)에서 핵산합성(核酸合成)에 미치는 영향(影響))

  • Seu, Yong-Taik
    • Applied Biological Chemistry
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    • v.21 no.2
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    • pp.84-102
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    • 1978
  • Barley embryoless half seeds were incubated in medium containing $10{\mu}M$ GA. Time course activity changes of ${\alpha}-amylase$ were studied in extract and medium seperately by the addition of $0.1{\mu}M,\;5{\mu}M,\;and\;10{\mu}M$ ABA in midcourse incubation of 10 hours after GA treatment. MAK profiles of nucleic acids in embryoless half seeds were compared either with $10{\mu}M$ GA treatment or concomitant treatment with $10{\mu}M$ GA and $10{\mu}M$ ABA after 10 hours incubation, Time course changes of weight increase, chlorophyll, protein and RNA consent in addition to RNase activity were studied in the presence of $10{\mu}M$ GA or $10{\mu}M$ ABA in barley coleoptile sections. After 20 hours incubation in the presence of plant hormones, MAK profiles of nucleic acids and reactive distribution of polysome and monosome were investigated. The above results were summarized as follows. 1) The production of ${\alpha}-amylase$ by treatment with GA alone increased at a linear rate in the incubation period and the active secretion of ${\alpha}-amylase$ began from 18 hours incubation in embryoless half seeds. 2) On the contrary to the partial inhibition by addition of $0.1{\mu}M$ ABA, the production of ${\alpha}-amylase$ was completely inhibited by both $5{\mu}M$ and $10{\mu}M$ ABA within 4 hours. Regardless of concentration of GA, the addition of $5{\mu}M$ ABA in midcourse completely inhibited the production of ${\alpha}-amylase$ 3) ABA treatment gave no effect on the secretion of ${\alpha}-amylase$. 4) There were no differences in RNA fractions between GA treatment and concomitant treatment with GA and ABA in the barlye embryoless half seeds. 5) While GA treatment increased the r-RNA fraction, ABA treatment decreased it and increased the s-RNA fraction in the coleoptile sections. 6) GA treatment increased RNA-DNA fraction best ABA treatment decreased it in the coleoptile sections. 7) While GA treatment suppressed RNase activity, ABA treatment increased it in the coleoptile sections. 8) GA treatment gave no great effect on the total RNA but ABA treatment remarkably diminished it in the coleoptile sections. 9) While GA treatment increased the growth and chlorophyll content, ABA treatment decreased them in the coleoptile sections. 10) GA treatment increased the protein synthesis and polysome formation but ABA treatment decreased them in the coleoptile sections. 11) The inhibition effect of ABA on polysome formation seemed to be resulted from the inhibition of r-RNA synthesis by ABA.

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Cell-Free miR-27a, a Potential Diagnostic and Prognostic Biomarker for Gastric Cancer

  • Park, Jong-Lyul;Kim, Mirang;Song, Kyu-Sang;Kim, Seon-Young;Kim, Yong Sung
    • Genomics & Informatics
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    • v.13 no.3
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    • pp.70-75
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    • 2015
  • MicroRNAs (miRNAs) have been demonstrated to play an important role in carcinogenesis. Previous studies revealed that miRNAs are present in human plasma in a remarkably stable form that is protected from endogenous RNase activity. In this study, we measured the plasma expression levels of three miRNAs (miR-21, miR-27a, and miR-155) to investigate the usefulness of miRNAs for gastric cancer detection. We initially examined plasma miRNA expression levels in a screening cohort consisting of 15 patients with gastric cancer and 15 healthy controls from Korean population, using TaqMan quantitative real-time polymerase chain reaction. We observed that the expression level of miR-27a was significantly higher in patients with gastric cancer than in healthy controls, whereas the miR-21 and miR-155a expression levels were not significantly higher in the patients with gastric cancer. Therefore, we further validated the miR-27a expression level in 73 paired gastric cancer tissues and in a validation plasma cohort from 35 patients with gastric cancer and 35 healthy controls. In both the gastric cancer tissues and the validation plasma cohort, the miR-27a expression levels were significantly higher in patients with gastric cancer. Receiver-operator characteristic (ROC) analysis of the validation cohort, revealed an area under the ROC curve value of 0.70 with 75% sensitivity and 56% specificity in discriminating gastric cancer. Thus, the miR-27a expression level in plasma could be a useful biomarker for the diagnosis and/or prognosis of gastric cancer.