• Journal of Microbiology and Biotechnology
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• 제15권5호
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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.

• 한국균학회지
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• 제14권3호
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• pp.201-207
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• 1986

효모 세포(S. uvarum)를 재료로 하여 배양 시기 및 sugar starvation시기에 특이하게 출현 또는 유도되는 RNase의 localization과 특성을 조사하고자 하였다. 정상 배양 시기 및 sugar starvation시킨 효모 세포를 세포 분획구에 따라 RNase 활성도를 측정하는 한편 ribosome 양의 변화를 조사하였다. 특히 ribosomal 분획구에서 추출한 RNase들을 poly(C)와 반응시킨 후 생성물을 TLC에 적응하여 효소의 특성 및 유도 여부를 조사하였다. 그 결과를 요약하면 다음과 같다. 세포 분획구 중 $45,000{\times}g$ pellet 분획구 및 Postribomosal 분획구에서는 배양 시기나 sugar starvation에 관계없이 RNase의 활성도는 유의하게 증감하지 않았으나, ribosomal 분회구에서는 정체기와 sugar starvation시 활성도가 각각 2배, 10배 이상 급격히 증가하였다. ribosome의 양적 동태를 살펴보면 early log phase의 세포에 비하여, 정체기 세포와 sugar starvation시킨 세포에서는 $1/3{\sim}1/6$까지 급격히 감소하였다. TLC의 결과 rRNase의 종류는 early log phase에서는 oligonuclease와 3'-ribonuclease, 5'-ribonuclease,stationarf phase에서는 oligonuclease, 3'-ribonuclease, sugar starvation 시켰을 때는 3'-ribonuclease, 5'-ribonuclease의 활성이 나타났다. 그리고 완전 배지를 사용한 효모 세포에서는 공통적으로 oligonuclease의 활성이 나타난 반면, sugar starvation시킨 효모 세포에서는 oligonuclease의 활성은 나타나지 않았다.

• Applied Biological Chemistry
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• 제20권2호
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• pp.242-246
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• 1977

보리(Hordeum vulgare cultivar Sedohadaka)초엽에 $1{\times}10^{-5}M$ Gibbrellic acid 및 $1{\times}10^{-5}M$ Abscisic acid를 처리(處理)하여 RNase활성(活性)의 경시적(經時的) 소장(消長)을 무처리구(無處理區)와 비교(比較)하고, 핵산(核酸)의 형태(形態)를 관찰하였으며 그 결과(結果)를 요약(要約)하면 다음과 같다. 1) GA는 RNase의 활성(活性)을 억제(抑制)시키는 반면(反面)ABA는 촉진(促進)시켰다. 2) 무처리구(無處理區)에서 정상적인 식물(植物)에 비(比)하여 r-RNA의 비(比)가 낮고 r-RNA의 비(比)가 높았는데 이것은 배양(培養)중 RNase의 작용(作用)에 의한 것 같다. 3) GA는 r-RNA의 분해(分解)를 완화시켰으나 ABA는 촉진(促進)시켰는데 이것은 RNase의 활성(活性)과 관계된 것 같다. 4) GA는 DNA-RNA 복합체(複合體)의 합성(合成)을 촉진(促進)시켰으나 ABA는 이를 억제시켰다. 5) ABA에 의한 s-RNA의 증가(增加)는 r-RNA의 분해산물(分解産物) 때문이라 생각된다.

• Animal cells and systems
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• 제2권4호
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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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• 제3권2호
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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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• 제15권2호
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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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• 제29권2호
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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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• 제21권2호
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• pp.84-102
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• 1978

대맥(大麥) 무배아반편종자(無胚芽半片種子)에 $10{\mu}M$ GA를 처리(處理)하여 배양(培養) 10시간(時間)에 배지(培地)에서 ABA의 농도(濃度)가 각각(各各) $0.1{\mu}M,\;5{\mu}M$ 및 $10{\mu}M$이 되도록 ABA 용액(溶液)을 첨가하여 ${\alpha}-amylase$ 생성(生成)의 소장(消長)을 그리고 $10{\mu}M$ GA와 $10{\mu}M$ ABA 혼합액(混合液) 처리(處理)하여 10시간(時間) 배양(培養) 후(後) 핵산(核酸)의 거동을 $10{\mu}M$ GA처리구(處理區)의 그것과 비교(比較)하였다. 대맥(大麥) 초엽(？葉)에 $10{\mu}M$ GA와 $10{\mu}M$ ABA를 처리(處理)하여 초엽의 생육(生育), chlorophyll, RNase의 활성(活性), 단백질(蛋白質) 및 총(繼) RNA 함량(含量)의 소장(消長)을 처리구(處理區)의 그들과 비교(比較)하였고 이들 hormone 처리(處理) 20시간(時間) 배양후(培養後) 핵산(核酸)의 거동 및 polysome과 menosome의 상대적(相對的) 분포(分布)를 조사(調査)하여 몇 가지 결론(結論)을 얻었으며 그 결과(結果)를 요약(要約)하면 다음과 같다. 1) GA 처리(處理)에 의(依)한 총(總) ${\alpha}-amylase$의 생성(生成)은 시간(時間)의 경과(經過)에 따라 직선적(直線的)으로 증가(增加)하였으며 ${\alpha}-amylase$의 분비(分泌)는 배양(培養) 18시간(時間)부터 활발(活潑)하였다. 2) $0.1{\mu}M$ ABA 첨가는 ${\alpha}-amylase$의 생성(生成)을 부분적(部分的)으로 저해(沮害)시켰으나 $5{\mu}M$ 및 $10{\mu}M$ ABA 첨가는 다같이 첨가 4시간(時間) 후(後)에 완전(完全)히 ${\alpha}-amylase$의 생성(生成)을 저해(沮害)시켰으며, 배양과정중(培養過程中) $5{\mu}M$ ABA 첨가는 GA의 농도(濃度)에 관계(關係)없이 완전(完全)히 ${\alpha}-amylase$의 생성(生成)을 저해(沮害)시켰다. 3) ABA는 무배아반편종자(無胚芽半片種子)에서 ${\alpha}-amylase$의 분비(分泌)에 별(別) 영향(影響)을 주지 않았다. 4) 무배아반편종자(無胚芽半片種子)에서 GA 단독(單獨) 처리구(處理區)와 GA-ABA 처리구간(處理區間)에 핵산(核酸)의 분획(分劃)에서 별(別) 이(異)가 없었다. 5) 초엽에서 GA의 처리(處理)는 r-RNA 획분(劃分)을 증가(增加)시킨 반면(反面) ABA 처리(處理)는 r-RNA 획분(劃分)을 감소(減少)시킴과 동시(同時)에 s-RNA 획분(劃分)을 증가(增加)시켰는권(權) 이는 이들 hormone이 RNase의 활성(活性)에 상이(相異)한 영향(影響)을 준 것으로 보였다. 6) 초엽에서 ABA 처리(處理)는 RNA-DNA 획분(劃分)의 성분비(成分比)를 감소(減少)시켰다. 7) 초엽에서 GA 처리(處理)는 RNase의 활성(活性)을 감소(減少)시켰으나 ABA 처리(處理)는 이의 활성(活性)을 증대(增大)시켰다. 8) 초엽에서 GA 처리(處理)는 총(總) RNA에 큰 영향(影響)을 주지 않았으나 ABA 처리(處理)는 이를 현저히 감소(減少)시켰다. 9) 초엽에 있어서 GA 처리(處理)는 초엽의 생장(生長) 및 chlorophyll 함량(含量)을 증가(增加)시켰으나 ABA 처리(處理)는 이들을 감소(減少)시켰다. 10) 초엽에서 GA 처리(處理)는 단백질(蛋白質) 및 polysome의 형성(形成)을 촉진(促進)시켰으나 ABA 처리(處理)는 이들을 감소(減少)시켰다. 11) ABA 처리(處理)는 polysome의 형성(形成)이 저해(沮害)되는 까닭은 ABA가 r-RNA의 합성(合成)을 저해(沮害)하는 것으로 보였다.

• BMB Reports
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• 제46권3호
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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.

• 대한본초학회지
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• 제21권2호
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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.