• 제목/요약/키워드: COP

검색결과 1,210건 처리시간 0.031초

Molecular Mechanism of Copper Resistance in Pseudomonas syringae pv. tomato.

  • Cha, Jae-Soon;Donald A. Cooksey
    • 한국식물병리학회:학술대회논문집
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    • 한국식물병리학회 1995년도 Proceedings of special lectures on Molecular Biological Approaches to Plant Disease National Agricultural Science and Technology Institute Suwon, Korea
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    • pp.97-117
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    • 1995
  • Copper resistance in Pseudomonas syringae pv. tomato is determined by copper-resistance operon (cop) on a highly conserved 35 kilobase plasmid. Copper-resistant strains of Pseudomonas syringae containing the cop operon accumulate copper and develop blue clonies on copper-containing media. The protein products of the copper-resistance operon were characterized to provide an understanding of the copper-resistance mechanism and its relationship to copper accumulation. The Cop proteins CopA (72 kDa), CopB (39 kDa), and CopC (12 kDa) were produced only under copper induction. CopA and CopC were periplasmic proteins and CopB was an outer membrane protein. Leader peptide sequences of CopA, CopB, and CopC were confirmed by amino-terminal peptide sequencing. CopA, CopB, and CopC were purified from strain PT23.2, and their copper contents were determined. One molecule of CopA bound 10.9${\pm}$1.2 atoms of copper and one molecule of CopC bound 0.6${\pm}$0.1 atom of copper. P. syringae cells containing copCD or copBCD cloned behind the lac promoter were hypersensitive to copper. The CopD (32 kDa), a probable inner membrane protein, function in copper uptake with CopC. The Cop proteins apparently mediate sequestration of copper outside of the cytoplasm as a copper-resistance mechanism.

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Aspergillus nidulans 분비소낭 구성요소인 α-COP과 ε-COP의 결합 부위 분석 (Analysis of Protein Domain for Interaction between α-COP and ε-COP in Aspergillus nidulans)

  • 송은정;김기현;이환희;박정석;강은혜;박희문
    • 한국균학회지
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    • 제40권4호
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    • pp.224-228
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    • 2012
  • A. nidulans ${\alpha}$-COP과 상호작용하는 단백질을 동정하기 위하여 ${\alpha}$-COP을 암호하는 유전자를 bait로 yeast two-hybrid 스크리닝용 A. nidulans cDNA 라이브러리를 탐색한 결과, COPI 소낭의 구성요소 중 하나인 ${\varepsilon}$-COP을 암호화하고 있는 유전자를 동정하고 $aneA^+$($\underline{A}$spergillus $\underline{n}$idulans $\underline{e}$psilon-COP, $AN{\varepsilon}$-COP)으로 명명하였다. $aneA^+$ 유전자는 총 296개의 아미노산을 암호화하고 있으며, 다른 균류의 ${\varepsilon}$-COP과 높은 상동성을 보였다. Yeast two hybrid 시스템으로 두 단백질 간의 상호작용 부위를 분석한 결과, ${\alpha}$-COP의 COOH 도메인과 ${\varepsilon}$-COP의 C-말단부가 필수 부위였으며, ${\alpha}$-COP N-말단의 WD 도메인과 ${\varepsilon}$-COP의 TPR 부위는 두 단백질 간의 결합을 촉진하는 조절부위로 밝혀졌다. 또한 사상균인 A. nidulans와 효모류인 S. cerevisiae에서 ${\alpha}$-COP과 ${\varepsilon}$-COP 간 작용양상이 유사한 것으로 보아, COPI 소낭의 구성요소인 ${\alpha}$-COP과 ${\varepsilon}$-COP 간의 상호작용 기전은 진핵세포 내에서 진화적으로 잘 보존되어 있는 것으로 추정되었다.

The Heavy Metal Tolerant Soil Bacterium Achromobacter sp. AO22 Contains a Unique Copper Homeostasis Locus and Two mer Operons

  • Ng, Shee Ping;Palombo, Enzo A.;Bhave, Mrinal
    • Journal of Microbiology and Biotechnology
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    • 제22권6호
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    • pp.742-753
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    • 2012
  • Copper-containing compounds are introduced into the environment through agricultural chemicals, mining, and metal industries and cause severe detrimental effects on ecosystems. Certain microorganisms exposed to these stressors exhibit molecular mechanisms to maintain intracellular copper homeostasis and avoid toxicity. We have previously reported that the soil bacterial isolate Achromobacter sp. AO22 is multi-heavy metal tolerant and exhibits a mer operon associated with a Tn21 type transposon. The present study reports that AO22 also hosts a unique cop locus encoding copper homeostasis determinants. The putative cop genes were amplified from the strain AO22 using degenerate primers based on reported cop and pco sequences, and a constructed 10,552 base pair contig (GenBank Accession No. GU929214). BLAST analyses of the sequence revealed a unique cop locus of 10 complete open reading frames, designated copSRABGOFCDK, with unusual separation of copCD from copAB. The promoter areas exhibit two putative cop boxes, and copRS appear to be transcribed divergently from other genes. The putative protein CopA may be a copper oxidase involved in export to the periplasm, CopB is likely extracytoplasmic, CopC may be periplasmic, CopD is cytoplasmic/inner membrane, CopF is a P-type ATPase, and CopG, CopO, and CopK are likely copper chaperones. CopA, B, C, and D exhibit several potential copper ligands and CopS and CopR exhibit features of two-component regulatory systems. Sequences flanking indicate the AO22 cop locus may be present within a genomic island. Achromobacter sp. strain AO22 is thus an ideal candidate for understanding copper homeostasis mechanisms and exploiting them for copper biosensor or biosorption systems.

인간 대장상피세포에서 항균펩타이드 CopA3에 의한 survivin 발현 조절 기작 규명 (Antimicrobial Peptide CopA3 Induces Survivin Expression in Human Colonocytes Through the Transcription Factor Sp1)

  • 김호
    • 생명과학회지
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    • 제32권1호
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    • pp.23-28
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    • 2022
  • 곤충에서 유래한 항균펩타이드 CopA3는 다양한 세포사멸 과정을 차단한다고 알려져 있다. 세균 톡신에 의한 상피세포 세포사멸이나 6-hydroxy dopamine이 야기하는 신경세포 세포사멸 모두를 차단한다. 연구자 등은 최근에 CopA3가 카스파제에 직접 결합하여 그들의 활성형-절단과정을 차단한다고 보고하였다. 하지만 강력한 CopA3의 항세포사멸 효능을 설명하기 위해서는 추가적인 규명이 필요한 실정이다. 본 연구에서는 세포사멸경로의 핵심억제인자인 survivin 발현에 미치는 CopA3의 영향을 확인하였다. 인간 대장상피세포(HT29)에 CopA3를 처리한 뒤 survivin 발현을 추적한 결과, survivin 단백질 양이 유의하게 증가함을 확인하였다. RT-PCR을 통해서 CopA3가 survivin 유전자의 전사를 증가시킴을 확인하였다. 그리고 CopA3 자극이 Sp1 발현을 증가시키는 사실과, Sp1 억제 물질인 tolfenamic acid 처리가 CopA3에 의한 survivin 증가를 차단한다는 결과들을 바탕으로 우리는 CopA3가 Sp1을 통해 survivin 발현을 유도한다는 최종 결론을 도출하였다. 한편 본 연구를 통해서 CopA3의 강력한 항세포사멸 효능을 설명할 수 있는 분자기작을 새롭게 제시하였다고 사려된다.

정적직립자세에서 여성고령자의 COP와 BBS, SPPB, TUG와의 상관관계연구 (Study of Correlation between BBS, SPPB, TUG and COP during Quiet Standing in Elderly Women)

  • 이경순;강영택
    • 한국운동역학회지
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    • 제19권3호
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    • pp.529-538
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    • 2009
  • 본 연구의 목적은 여성고령자의 균형과 관련된 평가항목 BBS(Berg balance scale), SPPB(short physical performance battery), TUG(timed up and go)를 측정하여 COP와의 상관관계를 알아보고자 하였다. 이를 위해 여성고령자 10명을 대상으로 신체기능검사와 정적직립자세 동안 지면경사(평지, 내리막, 오르막경사)에 따른 COP 이동범위와 평균속도를 산출하였다. 여성고령자들의 COP 좌우와 전후 값 비교에서 평지와 내리막, 오르막 모두 전후 값이 유의하게 크게 나타났으며, 좌우와 전후 각 방향에서 지면경사에 따른 유의한 차이는 나타나지 않았다. 신체 기능 점수와 COP 상관관계에서 BBS와 SPPB 점수가 높을수록 COP 이동범위와 평균속도 값은 감소하였으며 TUG 값이 클수록 COP 값이 크게 나타나는 유의한 상관관계를 나타냈다. 고령자들에게 균형과 관련된 신체기능검사와 COP는 지면경사에 따라 유의한 상관관계를 가지므로 임상에서 치료사들은 고령자들의 균형평가 시 COP 변수뿐만 아니라 BBS, SPPB, TUG를 통한 균형평가 활용이 가능하다고 본다.

Characterization of CaCOP1 Gene in Capsicum annuum Treated with Pathogen Infection and Various Abiotic Stresses

  • Guo, Jia;Seong, Eun-Soo;Wang, Myeong-Hyeon
    • Journal of Applied Biological Chemistry
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    • 제50권4호
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    • pp.227-233
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    • 2007
  • We characterized a full-length cDNA of CaCOP1 from pepper. Phylogenetic analysis based on the deduced amino acid sequence of CaCOP1 cDNA revealed high sequence similarity to the COP1 gene in Oryza sativa (84% identity). CaCOP1 shares high sequence identity with regulatory protein in Arabidopsis (84%), constitutively photomorphogenic 1 protein in Pisum sativum (81%) and COP1 homolog in Lycopersicon esculentum (79%). CaCOP1 gene exists single copy in the chili pepper genome. Expression of CaCOP1 was reduced in response to inoculation of non-host pathogens. The expression of this gene under abiotic and oxidative stresses was investigated, including 200 mM NaCl, 200 mM mannitol, cold ($4^{\circ}C$), 100 ${\mu}M$ abscisic acid (ABA), and 10 mM hydrogen peroxide ($H_2O_2$). CaCOP1 was induced significantly 3 h after low temperature treatment but not by dehydration or high salinity. Moreover, CaCOP1 was not induced by plant hormone ABA. These observations suggest that CaCOP1 gene plays a role in abiotic stress and may be belong to ABA-independent regulation system.

9-Meric Peptide Analogs of Defensin-like Antimicrobial Peptide Coprisin with Potent Antibacterial Activities with Bacterial Sell Selectivites

  • Shin, Areum;Lee, Eunjung;Kim, Jin-Kyoung;Bang, Jeong-Kyu;Kim, Yangmee
    • Bulletin of the Korean Chemical Society
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    • 제35권9호
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    • pp.2809-2812
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    • 2014
  • The 43-residue defensin-like peptide coprisin, which is isolated from dung bettle, Copris tripartitus, is a potent antimicrobial peptide. In our previous work, we determined the tertiary structure of coprisin and found that alpha helical region of coprisin from residue 19 to residue 30 is important for its antimicrobial activities. Here, we designed cop12mer and cop9mer analogs of coprisin based on the tertiary structure of coprisin. To investigate the relationship between hydrophobicity and antimicrobial activities and develop the potent peptide antibiotics, we designed cop9mer-1 with substitution of $His^2$ with Trp in cop9mer. The results showed that cop9mer-1 has higher toxicities as well as improved antimicrobial activities compared to cop9mer. In order to reduce the toxicity of cop9mer-1, we designed cop9mer-2 and cop9mer-3 with substitution of $Cys^3$ with Lys or Ser. Substitution of $Cys^3$ with these hydrophilic amino acids results in lower cytotoxicities compared to cop9mer-1. Cop9mer-2 with substitution of $Cys^3$ with Lys in Cop9mer-1 showed high antibacterial activities against drug resistant bacteria without cytotoxicity. Antibiotic action of cop9mer-1 analog appears to involve permeabilization of the bacterial cell membrane while cop9mer-2 and cop9mer-3 may have different mechanism of action. These results imply that that optimum balance in hydrophobicity and hydrophilicity in these 9-meric peptides plays key roles in their antimicrobial activities as well as cytotoxicities.

Cloning and Expression Characteristics of Pharbitis nil COP1 (PnCOP1) During the Floral Induction

  • 김윤희;김성룡;허윤강
    • Journal of Photoscience
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    • 제12권1호
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    • pp.1-9
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    • 2005
  • The ubiquitin E3 ligase COP1 (Constitutive Photomorphogenesis 1) is a protein repressor of photomorphogenesis in Arabidopsisplants, and it found in various organisms, including animals. The COP1 protein regulates the stability of many of the light-signaling components that are involved in photomorphogenesis and in the developmental processes. To study the effect of COP1 on flowering in a short day plant, we have cloned a full-length of PnCOP1 (Pharbitis nil COP1) cDNA from Pharbitis nil Choisy cv. Violet, and we examined its transcript levels under various conditions. A full-length PnCOP1 cDNA consists of 2,280 bp nucleotidesthat contain 47 bp of 5'-UTR, 232 bp of 3'-UTR including the poly (A) tail, and 1,998 bp of the coding sequence. The deduced amino acid sequence contains 666 amino acids, giving it a theoretical molecular weight of 75 kD and a isolectric point of 6.2. The PnCOP1 contains three distinct domains, an N-terminal $Zn^2+$-binding RING-finger domain, a coiled-coil structure, and WD40 repeats at the C-terminal, implying that the protein plays a role in protein-protein interactions. The PnCOP1 transcript was detected in the cotyledon, hypocotyls and leaves, but not in root. The levels of the PnCOP1 transcript were reduced in leaves that were a farther distance away from the cotyledons. The expression level of the PnCOP1 gene was inhibited by light, while the expression was increased in the dark. During the floral inductive 16 hour-dark period for Pharbitis nil, the expression was increased and it reached its maximum at the 12th hour of the dark period. The levels of PnCOP1 mRNA were dramatically reduced upon light illumination. These results suggest that PnCOP1 may play an important function in the floral induction of Pharbitis nil.

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전사 수준에서 repABC 유전자 발현을 조절하는 CopA 단백질의 역할 (Role of CopA to Regulate repABC Gene Expression on the Transcriptional Level)

  • 김삼웅;갈상완;지원재;방우영;김태완;백인규;방규호
    • 생명과학회지
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    • 제34권2호
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    • pp.86-93
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    • 2024
  • 플라스미드의 복제는 엄격하게 조절되어야 하기 때문에 일반적으로 rolling circle 복제를 수행하는 플라스미드들은 복제 개시인자인 RepB는 전사 및 번역 수준에서 엄격하게 조절되어 일정한 copy number를 유지한다. 플라스미드 pJB01에는 단일 오페론으로 구성된 세 개의 orfs (copA, repB, repC 또는 repABC)가 포함되어 있다. 아미노산 서열 분석에서 pJB01 CopA는 다른 플라스미드의 복제 수 조절 단백질로서 Cops와 상동성을 보였다. pMV158의 CopG와 비교할 때, CopA는 플라스미드의 일반화된 억제자의 모티브로 알려진 RHH (ribbon-helix-helix)를 형성하는 것으로 추정된다. gel mobility shift assay 결과 정제된 융합 단백질이 repABC 오페론의 operator 영역에 결합하는 것으로 나타났다. 전사 수준에 대한 CopA의 기능적 역할을 조사하기 위해 CopA R16M, K26R 및 E50V와 같은 세 개의 포인트 돌연변이가 CopA의 코딩 프레임에서 구성되었다. CopA R16M, K26R 및 E50V 돌연변이의 repABC mRNA 수준은 CopA wt보다 각각 1.84, 1.78 및 2.86배 증가했다. 또한 세 개의 CopA 유전자의 돌연변이로 인한 복제 수도 CopA wt보다 각각 1.86, 1.68 및 2.89배 증가했다. 이러한 결과는 CopA가 전사 억제자이며 복제 개시자로서 repABC mRNA 및 RepB 단백질 수를 감소시킴으로써 pJB01의 복제 수를 감소시키는 것으로 제의된다.

Dynamic Profile of the Copper Chaperone CopP from Helicobacter Pylori Depending on the Bound Metals

  • Hyun, Ja-shil;Park, Sung Jean
    • 한국자기공명학회논문지
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    • 제20권3호
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    • pp.76-81
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    • 2016
  • Copper is an elemental ion in living organisms. CopP from Helicobacter Pylori (HpCopP) is a copper(I)-binding protein and was suggested as regulator of copper metabolism in vivo. Previously, the metal binding property of HpCopP for Ag(I), Cu(I), and Cu(II) as well as the tertiary structure of HpCopP was shown. In this study, the dynamic profiles of HpCopP depending on metal binding were studied using ${^1H}-^{15}N$ steady-state NOE analysis. The heteroNOE experiment was performed for apo-CopP or metal-bound CopP. The obtained NOE values were analyzed and compared to figure out the effect of metals on the structural flexibility of HpCopP. As a result, Ag(I) and Cu(I) ions improved the rigidity of the structure while Cu(II) ion increased the flexibility of the structure, suggesting the oxidation of the CXXC motif decreases the structural stability of HpCopP.