• 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.

• 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.

• 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.

• 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.

• 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.

• 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.

• BMB Reports
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• v.29 no.2
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• pp.133-136
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• 1996

The affinity cleavage reagent Methidiumpropyl-EDTA-Iron(II) is applied to the structural analysis of 5S rRNA. Analysis of cleavage sites induced by MPE-Fe(II) on 5S rRNA shows that MPE intercalates easily between the unstable base pairs or into the bulges, thereby it strongly cuts the nucleosides nearby. The stable helical stems A, B, D and E as well as loop d are weakly cut. Most of the single-stranded loops are not cleaved. Based on the cleavage pattern of the 5S rRNA by MPE-Fe(II) and RNase V1, we suggest that MPE-Fe(II) may be used as a potential chemical probe in searching for the unstable helical regions of RNA, and for the sequences that appear to be involved in folding and distorting 5S rRNA.

• 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.

• 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.

• The Korea Journal of Herbology
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• v.21 no.2
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• pp.9-13
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• 2006

Objectives : To determine the DNA sequence of the 18S rRNA gene of the Rehmannia glutinosa and analyze it phylogenetically Methods : Dried root of the Rehmannia glutinosa was ground with a mortar and pestle. Glass beads(0.5 mm in diameter), TE buffer and SDS solution were added to that. The mixture was vortexed vigorously and extracted with the mixture of phenol, chloroform and isoamyl alcohol and with the mixture of the chloroform and isoamyl alcohol. The nucleic acids were precipitated with ethanol and resuspended in TE buffer. Contaminating RNA was digested with RNAse A and the DNA was purified further with the Geneclean Turbo Kit. This DNA was used as a template for amplification of the 18S rRNA gene by PCR. The PCR product was cloned in the pBluescript SK II plasmid by blunt-end ligation and the DNA sequence of the insert was determined. This DNA sequence was analyzed phylogenetically by the BLAST program. Results and Conclusion : Vortexing the ground powder of the dried plant root with glass beads during cell lysis improved recovery of DNA. The DNA sequence of the Rehmannia glutinosa 18S rRNA gene was determined and deposited at the GenBank as the accession number DQ469606. Phylogenetic analysis of that sequence showed the relationship between the members of the family of Scrophulariaceae and also the close relationship of the Buddleja davidii to the members of the Scrophulariaceae family.