• Title/Summary/Keyword: WD40

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Metacercarial proteins interacting with WD40-repeat protein of Clonorchis sinensis

  • Cho, Pyo-Yun;Kim, Tae-Im;Li, Shunyu;Hong, Sung-Jong;Choi, Min-Ho;Hong, Sung-Tae;Chung, Yong-Je
    • Parasites, Hosts and Diseases
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    • v.45 no.3
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    • pp.229-232
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    • 2007
  • The WD40-repeat proteins serve as a platform coordinating partner proteins and are involved in a range of regulatory cellular functions. A WD40-repeat protein (CsWD1) of Clonorchis sinensis previously cloned is expressed stage-specifically in the tegumental syncytium of C. sinensis metacercariae. In the present study, interact-ing proteins with the CsWD1 protein was purified by immunoprecipitation and 2 dimension gel electrophoresis from the C. sinensis metacercaria soluble extract, and tryptic peptides were analyzed by LC/ESI-MS. Putative partner proteins were annotated to be actin-2, glyceraldehyde-3-phosphate dehydrogenase, and hypothetical and unmanned proteins. The CsWD1 protein was predicted to contain 3 conserved actin-interacting residues on its functional surface. With these results, the CsWD1 protein is suggested to be an actin-interacting protein of C. sinensis.

A novel WD40 protein, BnSWD1, is involved in salt stress in Brassica napus

  • Lee, Sang-Hun;Lee, Jun-Hee;Paek, Kyung-Hee;Kwon, Suk-Yoon;Cho, Hye-Sun;Kim, Shin-Je;Park, Jeong-Mee
    • Plant Biotechnology Reports
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    • v.4 no.2
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    • pp.165-172
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    • 2010
  • Genes that are expressed early in specific response to high salinity conditions were isolated from rapeseed plant (Brassica napus L.) using an mRNA differential display method. Five PCR fragments (DD1.5) were isolated that were induced by, but showed different response kinetics to, 200 mM NaCl. Nucleotide sequence analysis and homology search revealed that the deduced amino sequences of three of the five cDNA fragments showed considerable similarity to those of ${\beta}$-mannosidase (DD1), tomato Pti-6 proteins (DD5), and the tobacco harpin-induced protein hin1 (DD4), respectively. In contrast, the remaining clones, DD3 and DD2, did not correspond to any substantial existing annotation. Using the DD3 fragment as a probe, we isolated a full-length cDNA clone from the cDNA library, which we termed BnSWD1 (Brassica napus salt responsive WD40 1). The predicted amino-acid sequence of BnSWD1 contains eight WD40 repeats and is conserved in all eukaryotes. Notably, the BnSWD1 gene is expressed at high levels under salt-stress conditions. Furthermore, we found that BnSWD1 was upregulated after treatment with abscisic acid, salicylic acid, and methyl jasmonate. Our study suggests that BnSWD1, which is a novel WD40 repeat-containing protein, has a function in salt-stress responses in plants, possibly via abscisic acid-dependent and/or -independent signaling pathways.

Validation of Reference Genes for Quantifying Changes in Physiological Gene Expression in Apple Tree under Cold Stress and Virus Infection (저온과 바이러스 감염에 노출된 사과나무의 생리적 유전자 정량 측정용 유전자들의 발현 분석 및 검증)

  • Yoon, Ju-Yeon;Jeong, Jae-Hoon;Choi, Seung-Kook
    • Research in Plant Disease
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    • v.26 no.3
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    • pp.144-158
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    • 2020
  • Quantitative reverse transcription PCR is used for gene expression analysis as the accurate and sensitive method. To analyze quantification of gene expression changes in apple plants, 10 housekeeping genes (ACT, CKL, EF-1α, GAPDH, MDH, PDI, THFs, UBC, UBC10, and WD40) were evaluated for their stability of expression during infection by Apple stem grooving virus (ASGV) or in cold-stress apple plant buds. Five reference-gene validation programs were used to establish the order of the most stable genes for ASGV as CKL>THFs>GAPDH>ACT, and the least stable genes WD40CKL>UBC10, and the least stable genes were ACT

Isolation and Analysis of the Yeast Mutant Gene, soo1-1, which Confers the Defect in $\beta$-1,3-glucan Biosynthesis (효모의 베타-1,3-글루칸 생합성 결함을 초래하는 돌연변이 유전자(soo1-1)의 분리 및 분석)

  • 이재준;이동원;김기현;박희문
    • Korean Journal of Microbiology
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    • v.37 no.1
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    • pp.42-48
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    • 2001
  • Allele rescue and sequence analysis of soo1-1 allele in Saccharomyces cerevisiae mutant LP0353 revealed that soo1-1 is identical to the previously reported ret1-1 allele, which has a base substitution of A for $G^{681}$ leading to an amino acid substitution of aspartic acid for $glycine^{227}$ in Soolp. However, it was revealed that the addition of osmotic stabilizer, such as 1.2M sorbitol can rescue the temperature sensitive phenotype of the ret1-1 mutant and that the soo1-1/ret1-1 mutation may confer defects in post-translational modification of proteins involved in the yeast cell wall biogenesis. Evidence for a putative role of 5th WD40 domain of the Soo1p/$\alpha$-COP in the construction and maintenance of cell walls was also presented by complementation test with deletion constructs of the SOOl.

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Determination of the MYB Motif Interacting with WD40 and Basic Helix Loop Helix Proteins

  • Kim, Ji-Hye;Kim, Bong-Gyu;Ahn, Joong-Hoon
    • Journal of Applied Biological Chemistry
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    • v.55 no.1
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    • pp.67-70
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    • 2012
  • Plant MYB transcription factors regulate secondary metabolism, cellular morphogenesis, and plant hormone signaling pathway. MYB proteins in plants consist of two repeats of 50 amino acid residues, which are referred to as R2R3 and they interact with WD40 or basic helix loop helix (bHLH) proteins. Yeast two hybrid assay was determined whether rice MYB protein interacts with either OsTTG1, which contains a WD40 domain, or with OsGL3, which contains a bHLH domain. Among 30 OsMYB proteins, three interacted with OsTTG1 and five interacted with OsGL3. A series of MYB mutants were created to determine the MYB domain important for the interaction with OsTTG1 or OsGL3. By using the yeast two hybrid assay, we found that the R3 motif of OsMYB10 and the R2 motif of OsMYB16 were required for interaction with OsTTG1 and OsGL3 proteins, respectively.

Pcp-2 Interacts Directly with Kinesin Superfamily KIF21A Protein (Kinesin superfamily KIF21A와 직접 결합하는 Pcp-2의 규명)

  • Park, Hye-Young;Kim, Sang-Jin;Ye, Sung-Su;Jang, Won-Hee;Lee, Sang-Kyeong;Park, Yeong-Hong;Jung, Yong-Wook;Moon, Il-Soo;Kim, Moo-Seong;Seog, Dae-Hyun
    • Journal of Life Science
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    • v.18 no.8
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    • pp.1059-1065
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    • 2008
  • KIF21A is a member of the Kinesin superfamily proteins (KIFs), which are microtubule-dependent molecular motors, anterograde axonal transporters of cargoes. Recently, congenital fibrosis of the extraocular muscles 1 (CFEOM1) has been shown to result from a small number of recurrent heterozygous missense mutations of KIF21A. CFEOM1 results from the inability of mutated KIF21A to successfully deliver cargoes to the development of the occulo-motor neuron or neuromuscular junction. Here, we used an yeast two-hybrid system to identify a protein that interacts with the WD-40 repeat domain of KIF21A and found a specific interaction with Purkinje cell protein-2 (Pcp-2), a small protein also known as L7. Pcp-2 protein bound to the WD-40 domain of KIF21A and KIF21B but not to other KIFs in yeast two-hybrid assays. In addition, this specific interaction was also observed in the glutathione S-transferase pull-down assay. An antibody to Pcp-2 specifically co-immunoprecipitated KIF21A associated with Pcp-2 from mouse brain extracts. These results suggest that Pcp-2 may be involved in the KIF21A-mediated transport as a KIF21A adaptor protein.

Assessment on the Extraction Efficiency of Explosive Compounds in Soil for Improving the Working Draft of International Standard (국제표준(안) 개선을 위한 토양 중 화약물질 추출 효율성 평가)

  • Lee, Goon-Taek;Jung, In-Ho;Kim, Bo-Hyun;Kim, Dong-Wook
    • Journal of Soil and Groundwater Environment
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    • v.17 no.1
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    • pp.42-46
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    • 2012
  • The ISO document of ISO/TC/190/SC3/WG11/N11 is a working draft of international standard (WD) dealing with analytical method for the determination of explosives and related compounds using high performance liquid chromatography. The scope of this WD covers the storage of samples, preparing test portion, extraction and instrumentation. The main purpose of this study was to improve the extraction conditions which were already adopted in the WD. For this purpose, mechanical shaking method could be corresponded up to 18 hours of ultrasonic bath extraction in the WD was tested. Methanol was also tested with the intention of being added as an extracting solvent other than acetonitrile in the WD. According to the results, 16 hours of mechanical shaking method showed statistically the same effectiveness as that of 18 hours of ultrasonic bath extraction. In case of extracting solvent, methanol also showed statistically the same extraction capability as acetonitrile for DNB, TNT, 2-A-DNT and 2,4-DNT. However, the recovery rate of TNB with methanol extraction was 40% higher than that of acetonitrile extraction. Through adding mechanical shaking method into committee draft (cf. the next stage draft of the WD during the process for making international standard), ISO standard of analyzing explosives and related compounds in soil would become more useful in dealing with huge number of field samples in the laboratory. In other aspect, adopting methanol as an alternative extracting solvent would be very effective in the terms of exchangeability with GC-ECD/MS method which is being developed by German experts.

Development of Loader Equipped with 4Wd and 4WS (II) (4WS System and Construction of Loader) (4WD 및 4WS이 가능한 로더 개발 (II) (4륜 조향장치 및 로더 구성))

  • 조현덕
    • Journal of the Korean Society of Manufacturing Technology Engineers
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    • v.8 no.1
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    • pp.150-157
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    • 1999
  • In this study, the loader was completed that has 4-wheel gear driven drivetrain of study (Ⅰ), the 4-wheel steering with power wheel type, all-wheel traction system, and joy-stick type lever for hydraulic control valve. From driving test of the developed 4WD and 4WS type loader, we obtained that the minimum circling radius and the necessary width in circling motion reduced about 40% and 33% compared with 2WS type loader. Also, all-wheel traction system could keep the tires glued to the ground with greater stability, the power steering allowed a smoother operation, and the joy-stick type lever offered easily to control. Thus, the developed loader having these functions was very fit in a small cattle shed or rugged ground.

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The Effects of Dietary Caloric Distribution on Insulin Secretion and Insulin Resistance in Sprague Dawley Rats (백서에서 식이내 열량 영양소의 배분이 인슐린 분비능과 인슐린 저항성에 미치는 영향)

  • Park, Sun-Min;Ahn, Seoung-Hee;Choi, Mi-Kyung;Kim, Young-Hee;Park, Chun-Hee;Choi, Soo-Bong
    • Journal of Nutrition and Health
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    • v.34 no.5
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    • pp.485-492
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    • 2001
  • The prevalence of type 2 diabetes mellitus has been rapidly increased in parallel with the westernization of eating behavior in Korea. Increased consumption of animal fat and simple sugar can be potential contributors for insulin resistance. The purpose of the study was to determine whether Western-(WD) and Korean-style(KD) diets altered insulin secretion and insulin resistance in male Sprague Dawley rats. Rats weighing 98$\pm$5g were provided by KD(77 En% of starch, 5 En% of corn oil and 13 En% of gluten plus 5 En% of casein), WD(42 En% of starch, 40 En% of butter and 18% of casein) or control diet(62 En% of starch, 20 En% of corn oil and 18% of casein) for 12 weeks. Body weights were lower in KD compared to WD. Fasting blood glucose levels were not different among diets. Insulin secretion from the beta cells was higher by 2.2$\pm$0.4 folds in WD than KD at baseline. In hyperglycemic clamp insulin secretion was higher in WD than KD and CD. Whole body glucose disposal rates referred to the state of insulin sensitivity were lowest in WD among groups. Glycogen deposits in soleus and quadriceps muscles were lowest in WD among all groups, but their triglyceride contents were highest. GLUT4 contents and glycogen synthase were lowest in WD in both muscles. In conclusions, westernization of diets needed more insulin to normalization of blood glucose levels due to increased insulin resistance. Thus, WD would lead to increased prevalence of diabetes mellitus when increased insulin resistance could not be compensated by insulin secretion in the case of elevated blood glucose levels. (Korean J Nutriton 34(5) : 485~492, 2001)

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Msi1-Like (MSIL) Proteins in Fungi

  • Yang, Dong-Hoon;Maeng, Shinae;Bahn, Yong-Sun
    • Mycobiology
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    • v.41 no.1
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    • pp.1-12
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    • 2013
  • Msi1-like (MSIL) proteins, which are eukaryote-specific and contain a series of WD40 repeats, have pleiotropic roles in chromatin assembly, DNA damage repair, and regulation of nutrient/stress-sensing signaling pathways. In the fungal kingdom, the functions of MSIL proteins have been studied most intensively in the budding yeast model Saccharomyces cerevisiae, an ascomycete. Yet their functions are largely unknown in other fungi. Recently, an MSIL protein, Msl1, was discovered and functionally characterized in the pathogenic yeast Cryptococcus neoformans, a basidiomycete. Interestingly, MSIL proteins appear to have redundant and unique roles in both fungi, suggesting that MSIL proteins may have evolutionarily divergent roles in different parts of the fungal kingdom. In this review, we will describe the current findings regarding the role of MSIL proteins in fungi and discuss future directions for research on this topic.