• Journal of Microbiology
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• 제44권5호
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• pp.508-514
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• 2006

In this study, the double-stranded DNA-dependent activities of Deinococcus radiodurans RecA protein (Dr RecA) were characterized. The interactions of the Dr RecA protein with double-stranded DNA were determined, especially dsDNA-dependent ATP hydrolysis by the Dr RecA protein and the DNA strand exchange reaction, in which multiple branch points exist on a single RecA protein-DNA complex. A nucleotide cofactor (ATP or dATP ) was required for the Dr RecA protein binding to duplex DNA. In the presence of dATP, the nucleation step in the binding process occurred more rapidly than in the presence of ATP. Salts inhibited the binding of the Dr RecA protein to double-stranded DNA. Double-stranded DNA-dependent ATPase activities showed a different sensitivity to anion species. Glutamate had only a minimal effect on the double-stranded DNA-dependent ATPase activities, up to a concentration of 0.7 M. In the competition experiment for Dr RecA protein binding, the Dr RecA protein manifested a higher affinity to double-stranded DNA than was observed for single-stranded DNA.

• 자원ㆍ환경경제연구
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• 제29권1호
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• pp.91-111
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• 2020

신재생에너지 발전 규모 확대를 위해 신재생에너지공급의무화(RPS) 제도가 실시되는 가운데, 발전사업자는 신재생에너지 공급인증서(REC)를 확보하여 이를 시설 운영에 대한 인센티브로 활용할 수 있다. 태양광 발전을 통해 확보된 REC는 현물시장 또는 고정가격계약을 통해 거래될 수 있으며, 현물시장 거래 시 발전사업자는 REC 현물시장 가격의 불확실성에 노출된다. 본 연구에서는 REC 현물시장 가격의 불확실성을 고려하여 태양광 발전사업자의 REC 거래 방식 최적 전환 시점을 분석하기 위해 실물옵션 분석을 실시한다. 분석을 통해 REC 거래 방식을 현물시장 거래에서 고정가격계약 거래로 전환할 수 있는 REC 임계 가격을 산출하였다. 민감도 분석 결과 REC 현물시장 가격의 불확실성을 고려한 경우에는 현물시장 거래가 합리적 거래 방식으로, 불확실성을 고려하지 않은 경우에는 고정가격계약 거래가 합리적 거래 방식으로 나타났다.

• 미생물학회지
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• 제27권2호
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• pp.162-167
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• 1989

Western blot analysis of lysates of Streptococcus pneumoniae revealed a single polypeptide species that cross-reacted with E. coli RecA antiserum. The apparent molecular weight of this putative RecA protein analog (RecAsp) was 24, 000 smaller than any other known RecA analogue. The RecAsp protein was present at the same level in competent and non-competent cells.

• 미생물학회지
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• 제43권4호
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• pp.250-255
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• 2007

Deinococcus radiodurans recA는 이 미생물의 방사선 저항성을 나타내는 표현형에 필수적이며 재조합성 DNA 수선 과정에 관여한다. 이 과정에서 RecA 단백질은DNA와 결합하여 반응의 활성 종인 RecA nucleoprotein 필라멘트를 형성한다. DNA-의존성 ATPase 활성과 함께, RecA 단배질의 외가닥 DNA 혹은 이중가닥 DNA와의 상호작용은 RecA 단백질이 관여하는 반응의 중심과정으로 이에 관한 분석을 시도하였다. D. radiodurans RecA 단배질은 DNA에 결합한 DNA-단백질 복합체만이 ATPase 활성을 나타내므로, ATP (혹은 dATP) 가수분해를 측정함으로써 RecA와 외가닥 DNA와의 상호작용 정도를 분석하였다. D. radiodurans RecA 단백질은 외가닥 DNA의 염기 구성의 이질성에 영향을 받았으며, homopolymer인 poly(dT)와의 상호작용에서 가장 높은 가수분해 활성을 보였다. Homopolymer인 합성 DNA-의존성 ATP 및 dATP의 가수분해는 pH 6.0과 9.0의 범위에서 다소 일정한속도로 일어났으며 최적 pH는 7.0과 7.5 사이였다. 외가닥 DNA-의존성 ATPase 활성은 염의 존재에 영향을 받아 KCl이 존재하면 다소 억제되나, K-glutamate가 존재하면 오히려 촉진되었다. RecA 단백질과 외가닥 DNA의 상호작용을 ATP 가수분해로 분석하였을 때 2 mM 이상의 magnesium 이온이 DNA 결합반응에 필요하였으며, 비교적 넓은 범위의 pH에서 외가닥 DNA와의 결합반응이 일어나며, 이러한 결합반응은 당량적인 비(1:3, RecA protein: DNA nucleotide)로 일어났다.

• Fisheries and Aquatic Sciences
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• 제25권1호
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• pp.12-19
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• 2022

In the present study, we investigated the effects of recombinant eel gonadotropins (rec-GTHs) on maturation induction in immature ovarian culture in vitro and sex steroid hormones in vivo in the Japanese eel Anguilla japonica. To study the in vitro effects of rec-GTHs on estradiol-17β (E2) production in immature ovarian tissues, ovarian tissues were incubated with different doses of rec-follicle-stimulating hormone (rec-FSH) or rec-luteinizing hormone (rec-LH). The results revealed that the E2 levels in the rec-FSH (0.1, 0.5, or 1 ㎍/mL)- and rec-LH (0.1 or 0.5 ㎍/mL)-treated groups were significantly higher than those in the female eels from the control group. Furthermore, to investigate the in vivo effects of rec-GTHs on the gonadosomatic index (GSI) and plasma sex steroid hormone levels, the eels were injected intraperitoneally with eel's ringer (control), salmon pituitary extract (SPE; for female eels), human chorionic gonadotropin (hCG; for male eels), rec-FSH, rec-LH, and rec-FSH + rec-LH once a week. The results revealed that except for the SPE and the hCG groups, none of the groups exhibited a significant difference in GSI values. However, in vivo plasma E2 levels increased at the end of 4 weeks after rec-FSH treatment in female eels. Based on these results, it is suggested that rec-GTHs may have a positive effect on sexual maturation in female eels; however, further studies on complementary rec-protein production systems and additional glycosylation of rec-hormones are needed to elucidate hormone bioactivity in vivo and in vitro.

• 한국동물학회지
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• 제37권2호
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• pp.232-239
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• 1994

RecA protein plans a central role in homologous recombination and DNA repair in Escherichia cofi (E. colD. The function 8nd structure of this protein are universal in prokarvotes and also conserved in eukaryotes such as yeast. The RecA-like protein with 74 lInDa in size has already been identified and purified from a fission yeast Schizosaccharomyces pombe (5. pommel (Lee, 19911. From this study it was revealed that the RecA-like protein of 5. pombe was highly inducible to various DNA damaging agents and inhibitors of nucleotide pool svnthesizins enzymes. The cellular level of the 5. pombe RecA-like protein wi,u markedly increased, upto 5- to 10-fold, by treatment with various DNA-damains agents including ultraviolet (UV) light, methyl methanesulfonate WS),4-nitroquinoline-1-oxide (4-NQO), and mitomycin-C (MMC), similar to E. cofi RecA protein. Interestingly, the protein level was also increased by inhibitors of nucleotide pool forming enzlwnes such as methotrexate (MTX) and hvdroxvurea (HU). The most effective doses for the inducibility of 4-NQO, MMS, W, MMC, MTX, and HU were 0.2 Ug/ml, 30 mM, 200 J/ma, 0.4 $\mus/ml,$ 1 Ug/ml, and 100 mM, respectively. The range of effective duration time for the inducibilitv of RecA-like protein was from 270 to 450 mins. These results suggest that the 5. pombe RecA-like protein also platys an imortant role in cellular responses to DNA damage as in E. coli system.

• Journal of Radiation Protection and Research
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• 제20권2호
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• pp.71-78
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• 1995

본 연구에서는 유전자 손상회복에 관여하는 단백질을 이용하여 돌연변이 생성을 억제시키는 물질로서 알려진 cobaltous chloride가 유전자 손상회복에 미치는 영향을 연구하므로서 방사선으로 인한 손상방지 및 방사선 방어효과에 대한 적용가능성을 평가하였다. Cobaltous chloride가 RecA 단백질의 기능에 미치는 영향을 조사한 결과 RecA 단백질에 의한 DNA strand exchange 반웅에 있어 cobaltous chloride 처리로 RecA 단백질이 $_{ss}DNA$로 부터 SSB 단백질과 더 효과적으로 경쟁함으로써 안정된 $RecA-_{ss}DNA$ complex의 형성을 유도하고, 증가된 ATPase활성에 의한 ATP 가수분해로 손상된 DNA의 회복이 촉진될 수 있다는 사실을 입증 해주고 있다. 또한 RecA단백질은 UV에 의해 손상된 supercoiled DNA에 더 효과적으로 결합됨이 관찰되었으며 UV 선량과도 상관관계가 있음을 확인하였다. 따라서 이와 같은 연구결과들은 방사선으로 인한 유전적인 손상방지 및 방사선 방어효과에 관한 연구에 적용될 수 있을 것으로 기대된다.

• Journal of Microbiology
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• 제37권4호
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• pp.248-255
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• 1999

We have analyzed the MRE3/REC114 gene of Saccharomyces cerevisiae, previously detected in isolation of mutants defective in meiotic recombination. We cloned the MRE3/REC114 gene by complementation of the meiotic recombination defect and it has been mapped to chormosome XIII. The DNA sequence analysis revealed that the MRE3 gene is identical to the REC114 gene. The upstream region of the MRE3/REC114 gene contains a T_4C site, a URS (upstream repression sequence) and a TR (T-rich) box-like sequence, which reside upstream of many meiotic genes. Coincidentally, northern blot analysis indicated that the three sizes of MRE3/REC114 transcripts, 3.4, 1.4 and 1.2 kb, are induced in meiosis. A less abundant transcript of 1.4 kb is detected in both mitotic and meiotic cells, suggesting that it is needed in mitosis as well as meiosis. To examine the role of the MRE3/REC114 gene, we constructed mre3 disruption mutants. Strains carrying an insertion or null deletion of the MRE3/REC114 gene showed slow growth in nutrient medium and the doubling time of these cells increased approximately by 2-fond compared to the wild-type strain. Moreover, the deletion mutant (${\delta}$mre3) displayed no meiotically induced recombination and no viable spores. The mre3/rec114 spore lethality can be suppressed by spo13, a mutation that causes cells to bypass reductional division. The double-stranded breaks (DSBs) which are involved in initiation of meiotic recombination were not detected in the analysis of meiotic chromosomal DNA from the mre3/rec114 disruptant. From these results we suggest that the MRE3/REC114 gene product is essential in normal growth and in early meiotic stages involved in meiotic recombination.

• Animal cells and systems
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• 제3권2호
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• pp.229-236
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• 1999

The recA gene plays a central role in genetic recombination and SOS DNA repair in Escherichia coli (E. coli). We have previously identified a 42 kDa RecA-like protein inducible by a variety of DNA damages from a fluorescent Pseudomonas strain sp. and characterized its inducible kinetics. In the present study, we cloned and characterized the gene encoding the RecA-like protein by immunological screening of Pseudomonas genomic expression library using polyclonal E. coli anti-RecA antibodies as a probe. From 10$^{5}$ plaques screened, five putative clones were finally isolated. Southern blot analysis indicated that four clones had the same DNA inserts and the recA-like gene was located within the 3.2 kb EcoRI fragment of Pseudomonas chromosomal DNA. In addition, the cloned recA-like gene was transcribed into an RNA transcript approximately 1.1 kb in size, as judged by Northern blot analysis. The cellular level of RNA transcript of the cloned recA-like gene was increased to an average of 5.15- fold upon treatment with DNA damaging agents such as ultraviolet (UV)- light, nalidixic acid (NA), methyl methanesulfonate (MMS), and mitomycin-C (MMC). These results suggest that the cloned gene is inducible by DNA damage similarly to the recA gene in E. coli. However, the cloned gene did not restore the DNA damage sensitivity of the E. coli recA-mutant.

• 미생물학회지
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• 제46권4호
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• pp.313-318
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• 2010

D. radiodurans RecA 단백질은 DNA에 결합한 DNA-단백질 복합체만이 ATPase 활성을 나타내며. 보통의 낮은 염 농도조건에서는 DNA가 존재하지 않으면 RecA 단백질에 의한 ATP 가수분해는 거의 일어나지 않았으나 이러한 ATP 가수분해현상은 높은 농도의 염을 첨가하게 되면 1,000배 활성화 되었으며 1.6 M KCl이 존재할 때 ATP 혹은 dATP를 가수분해 하였다. DNA가 존재하지 않을 때 염에 의해 촉진되는 활성은 RecA 단백질 농도에 비례하였고, 더 높은 염농도에서 더 높은 ATP 가수분해 활성이 나타났다. 이러한 활성화 현상을 다양한 종류의 이온 형태에서 분석하였을 때 1.6 M Cl 음이온이 존재할 때 양이온의 형태에 따른 활성화 정도는 $K^+{\geq}Na^+$> $NH_4^+$의 경향을 보였으며, 1.6 M의 K 양이온 존재할 때 음이온의 형태에 따른 활성화는 glutamate > $Cl^-$ > acetate > $PO_4^-$의 순서로 높게 나타났다. 고농도의 염이 존재하는 조건에서 DNA 비의존성 ATPase의 활성은 비교적 넓은 범위 최적 조건인 pH7과 pH 8 사이에서 최대 활성을 보였고, 기질에 대한 친화도면에서도 외가닥 DNA 의존성 활성보다는 이중가닥 DNA 의존성 활성형태를 보였다. 고농도의 염이 첨가되고 DNA가 존재하지 않을 때 RecA 단백질에 의한 ATP 가수분해를 위한 RecA 단백질의 활성 종 형태는 최소 3개의 RecA 단백질이 결합되어 있는 과량체로 작용하는 것으로 나타났다.