• Title/Summary/Keyword: COP1

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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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    • v.50 no.4
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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.

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

  • Kim, Yun-Hee;Kim, Seong-Ryong;Heo, Yoon-Kang
    • Journal of Photoscience
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    • v.12 no.1
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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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Molecular Mechanism of Copper Resistance in Pseudomonas syringae pv. tomato.

  • Cha, Jae-Soon;Donald A. Cooksey
    • Proceedings of the Korean Society of Plant Pathology Conference
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    • 1995.06b
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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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Antimicrobial Peptide CopA3 Induces Survivin Expression in Human Colonocytes Through the Transcription Factor Sp1 (인간 대장상피세포에서 항균펩타이드 CopA3에 의한 survivin 발현 조절 기작 규명)

  • Kim, Ho
    • Journal of Life Science
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    • v.32 no.1
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    • pp.23-28
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    • 2022
  • CopA3 (LLCIALRKK), an antimicrobial peptide isolated from the Korean dung beetle, has been shown to suppress apoptosis in various cell types. CopA3 inhibits not only bacterial toxin-induced colonocyte apoptosis but also 6-hydroxy dopamine-induced neural cell apoptosis. Our recent study revealed that CopA3 directly binds to caspases (key regulators of apoptosis) and inhibits the proteolytic cleavage required for their activation. But molecular mechanisms underlying CopA3-mediated inhibition of apoptosis in multiple cell types remain unknown. Here we assessed possible effects of CopA3 on expression of survivin, which is known to inhibit apoptosis. In HT29 human colonocytes, CopA3 exposure markedly upregulated survivin expression in a concentration- and time-dependent manner. RT-PCR revealed that CopA3-mediated upregulation of survivin was attributable to increased gene transcription, and further showed that CopA3 also increased expression of Sp1, one of many transcription factors known to be involved in transcription of the survivin gene. Notably, blocking Sp1 by treatment with the Sp1 inhibitor, tolfenamic acid, significantly reduced CopA3-mediated upregulation of survivin. These results collectively suggest that CopA3 induces Sp1 expression, which in turn is involved in upregulation of survivin in human colonocytes. These novel findings establish another pathway for explaining the anti-apoptotic effects of CopA3 against various cellular apoptosis systems.

Photoperiod sensing system for timing of flowering in plants

  • Lee, Byoung-Doo;Cha, Joon-Yung;Kim, Mi Ri;Paek, Nam-Chon;Kim, Woe-Yeon
    • BMB Reports
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    • v.51 no.4
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    • pp.163-164
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    • 2018
  • CONSTANS (CO) induces the expression of FLOWERING LOCUS T (FT) in the photoperiodic pathway, and thereby regulates the seasonal timing of flowering. CO expression is induced and CO protein is stabilized by FLAVIN-BINDING KELCH REPEAT F-BOX PROTEIN 1 (FKF1) in the late afternoon, while CO is degraded by CONSTITUTIVE PHOTOMORPHOGENIC 1 (COP1) during the night. These regulatory cascades were thought to act independently. In our study, we investigated the relationship between FKF1 and COP1 in the regulation of CO stability in response to ambient light conditions. A genetic analysis revealed that FKF1 acts as a direct upstream negative regulator of COP1, in which cop1 mutation is epistatic to fkf1 mutation in the photoperiodic regulation of flowering. COP1 activity requires the formation of a hetero-tetramer with SUPPRESSOR OF PHYA-105 (SPA1), [$(COP1)_2(SPA1)_2$]. Light-activated FKF1 has an increased binding capacity for COP1, forming a FKF1-COP1 hetero-dimer, and inhibiting COP1 homo-dimerization at its coiled-coil (CC) domain. Mutations in the CC domain result in poor COP1 dimerization and misregulation of photoperiodic floral induction. We propose that FKF1 represses COP1 activity by inhibiting COP1 dimerization in the late afternoon under long-day conditions, resulting in early flowering.

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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    • v.35 no.9
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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.

Effects of support feet on active feet during Y-Balance Test (Y-Balance Test 시 지지발이 활동발에 미치는 영향)

  • Byung-Hoon Woo
    • Journal of the Korean Applied Science and Technology
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    • v.40 no.6
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    • pp.1249-1258
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    • 2023
  • The purpose of this study was to reveal the effect of COP variable of support feet on relative reach distance and composite score of active feet during YBT. 29 adults in their 20s(age: 24.4±3.0 yrs, height: 171.0±10.5 cm, weight: 72.1±12.1 kg, leg length: 88.2±5.8 cm) participated in the study. Using YBT, relative reach distance and composite score of active feet were calculated, and COP variables of support feet were measured on left and right support feet. Multiple regression analysis was used to determine the effect of COP variables of support feet during YBT on relative reach distance and composite score. As a result of the study, during YBT, when supporting right feet, the influence of AP COP velocity, ML COP velocity and COP velocity in posterolateral direction was significant, and when supporting left feet, the influence of AP COP velocity, ML COP velocity in posteromedial direction improved relative reach distance and composite score.

Effect of Zanthoxylum piperitum Extract on Human Skin Protection from UVB by Regulation of COP1 and PPAR-α (초피나무 열매 추출물의 COP1 및 PPAR-α 조절을 통한 자외선에 대한 피부 보호 효과)

  • Kim, Yun-Sun;Kim, Yumi;Lee, Sanghwa
    • Journal of the Society of Cosmetic Scientists of Korea
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    • v.42 no.4
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    • pp.393-401
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    • 2016
  • Ultraviolet (UV) irradiation from the sun is the primary environmental factor that causes skin damages including skin cancer and premature skin aging. Because, even the most powerful sunscreen can't always afford enough protection, it is necessary to enhance the defensive power of skin against UV. Recently, constitutive photomorphogenic protein-1 (COP1) has shown to contribute to the regulation of UVB response of keratinocytes. In this study, we represent that COP1 and its associated protein, de-etiolated 1 (DET1), might participate in photoaging process in human skin as Arabidopsis COP1 does sun-protective function in plants. After UVB irradiation, the decrease of COP1 and DET1 mRNA expression was followed by the increase of c-Jun total protein. Moreover, transfection with DNA vectors expressing COP1 and DET1 down-regulated the c-Jun total protein. We found that Zanthoxylum piperitum extract (ZE) up-regulated the expression of COP1 and DET1 on human keratinocytes, and inhibited the expression of MMP1 which is one of the genes regulated by c-Jun signal. In addition, ZE has been reported to stimulate PPAR-${\alpha}$ and strengthen the skin barrier. We found that ZE decreased the UVB-induced IL-6 and IL-8 in NHEK cells. In human study, ZE protected skin against UV-B induced erythema and erythema-induced pigmentation. These results indicate that ZE could be useful for the protection against the adverse effects of UV irradiation through various mechanisms.

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

  • Sam Woong Kim;Sang Wan Gal;Won-Jae Chi;Woo Young Bang;Tae Wan Kim;In Gyu Baek;Kyu Ho Bang
    • Journal of Life Science
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    • v.34 no.2
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    • pp.86-93
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    • 2024
  • Since replication of plasmids must be strictly controlled, plasmids that generally perform rolling circle replication generally maintain a constant copy number by strictly controlling the replication initiator Rep at the transcriptional and translational levels. Plasmid pJB01 contains three orfs (copA, repB, repC or repABC) consisting of a single operon. From analysis of amino acid sequence, pJB01 CopA was homologous to the Cops, as a copy number control protein, of other plasmids. When compared with a CopG of pMV158, CopA seems to form the RHH (ribbon-helix-helix) known as a motif of generalized repressor of plasmids. The result of gel mobility shift assay (EMSA) revealed that the purified fusion CopA protein binds to the operator region of the repABC operon. To examine the functional role of CopA on transcriptional level, 3 point mutants were constructed in coding frame of copA such as CopA R16M, K26R and E50V. The repABC mRNA levels of CopA R16M, K26R and E50V mutants increased 1.84, 1.78 and 2.86 folds more than that of CopA wt, respectively. Furthermore, copy numbers owing to mutations in three copA genes also increased 1.86, 1.68 and 2.89 folds more than that of copA wt, respectively. These results suggest that CopA is the transcriptional repressor, and lowers the copy number of pJB01 by reducing repABC mRNA and then RepB, as a replication initiator.

Effects of increasing weight load on dynamic balance (무게부하 증가가 동적 균형에 미치는 영향)

  • Woong-Jae Choi;Byung-Hoon Woo
    • Journal of the Korean Applied Science and Technology
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    • v.40 no.4
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    • pp.606-615
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    • 2023
  • The purpose of this study was to investigate the effect of increasing weight load on the dynamic balance on Y-balance Test(YBT). 18 male and female adults in their 20-30s (age: 23.17±1.72 years, height: 172.46±9.84 cm, weight: 73.39±11.44 kg, leg length: 88.89±5.69 cm) participated in the experiment. To measure the dynamic balance, the YBT was used to measure the reach distance, composite score, and COP variables to derive the results. As a result of the study, dynamic balance was improved in weight load in posteromedial and posterolateral reach and overall score during YBT. As a result of COP, anteroposterior amplitude, left and right COP velocity, and COP area during anterior reach improved dynamic balance in weight load in the right foot, but there was no difference during the posteromedial reach, ML amplitude in the left foot, AP COP velocity in left foot, ML COP velocity and COP velocity in both foot, and COP area in both foot improved dynamic balance when weight load during posterolateral reach.