• Title/Summary/Keyword: Molecular evolution

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KBTBD7, a novel human BTB-kelch protein, activates transcriptional activities of SRE and AP-1

  • Hu, Junjian;Yuan, Wuzhou;Tang, Ming;Wang, Yuequn;Fan, Xiongwei;Mo, Xiaoyang;Li, Yongqing;Ying, Zaochu;Wan, Yongqi;Ocorr, Karen;Bodmer, Rolf;Deng, Yun;Wu, Xiushan
    • BMB Reports
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    • v.43 no.1
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    • pp.17-22
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    • 2010
  • In this study, a novel member of BTB-kelch proteins, named KBTBD7, was cloned from a human embryonic heart cDNA library. The cDNA of KBTBD7 is 3,008 bp long and encodes a protein product of 684 amino acids (77.2 kD). This protein is highly conserved in evolution across different species. Western blot analysis indicates that a 77 kD protein specific for KBTBD7 is wildly expressed in all embryonic tissues examined. In COS-7 cells, KBTBD7 proteins are localized to the cytoplasm. KBTBD7 is a transcription activator when fused to GAL4 DNA-binding domain. Deletion analysis indicates that the BTB domain and kelch repeat motif are main regions for transcriptional activation. Overexpression of KBTBD7 in MCF-7 cells activates the transcriptional activities of activator protein-1 (AP-1) and serum response element (SRE), which can be relieved by siRNA. These results suggest that KBTBD7 proteins may act as a new transcriptional activator in mitogen-activated protein kinase (MAPK) signaling.

Micro/Nanotribology and Its Applications

  • Bhushan, Bharat
    • Tribology and Lubricants
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    • v.11 no.5
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    • pp.128-135
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    • 1995
  • Atomic force microscopy/friction force microscopy (AFM/FFM) techniques are increasingly used for tribological studies of engineering surfaces at scales, ranging from atomic and molecular to microscales. These techniques have been used to study surface roughness, adhesion, friction, scratching/wear, indentation, detection of material transfer, and boundary lubrication and for nanofabrication/nanomachining purposes. Micro/nanotribological studies of single-crystal silicon, natural diamond, magnetic media (magnetic tapes and disks) and magnetic heads have been conducted. Commonly measured roughness parameters are found to be scale dependent, requiring the need of scale-independent fractal parameters to characterize surface roughness. Measurements of atomic-scale friction of a freshly-cleaved highly-oriented pyrolytic graphite exhibited the same periodicity as that of corresponding topography. However, the peaks in friction and those in corresponding topography were displaced relative to each other. Variations in atomic-scale friction and the observed displacement has been explained by the variations in interatomic forces in the normal and lateral directions. Local variation in microscale friction is found to correspond to the local slope suggesting that a ratchet mechanism is responsible for this variation. Directionality in the friction is observed on both micro- and macro scales which results from the surface preparation and anisotropy in surface roughness. Microscale friction is generally found to be smaller than the macrofriction as there is less ploughing contribution in microscale measurements. Microscale friction is load dependent and friction values increase with an increase in the normal load approaching to the macrofriction at contact stresses higher than the hardness of the softer material. Wear rate for single-crystal silicon is approximately constant for various loads and test durations. However, for magnetic disks with a multilayered thin-film structure, the wear of the diamond like carbon overcoat is catastrophic. Breakdown of thin films can be detected with AFM. Evolution of the wear has also been studied using AFM. Wear is found to be initiated at nono scratches. AFM has been modified to obtain load-displacement curves and for nanoindentation hardness measurements with depth of indentation as low as 1 mm. Scratching and indentation on nanoscales are the powerful ways to screen for adhesion and resistance to deformation of ultrathin fdms. Detection of material transfer on a nanoscale is possible with AFM. Boundary lubrication studies and measurement of lubricant-film thichness with a lateral resolution on a nanoscale have been conducted using AFM. Self-assembled monolyers and chemically-bonded lubricant films with a mobile fraction are superior in wear resistance. Finally, AFM has also shown to be useful for nanofabrication/nanomachining. Friction and wear on micro-and nanoscales have been found to be generally smaller compared to that at macroscales. Therefore, micro/nanotribological studies may help def'me the regimes for ultra-low friction and near zero wear.

Cloning of Pig Kidney cDNA Encoding an Angiotensin I Converting Enzyme (돼지 신장의 Angiotensin I Converting Enzyme cDNA 클로닝)

  • Yoon, Jang-Ho;Yoon, Joo-Ok;Hong, Kwang-Won
    • Applied Biological Chemistry
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    • v.49 no.4
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    • pp.293-297
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    • 2006
  • Angiotensin converting enzyme(ACE) is a zinc-containing dipeptidase widely distributed in mammalian tissues and is thought to play a significant role in blood pressure regulation by hydrolyzing angiotensin I to the potent vasoconstrictor, angiotensin II. Recently, the presence of ACE in pig ovary was reported and the ACE from pig kidney was isolated and characterized. However no nucleotide sequence of the ACE gene from pig is yet known. We report here the cloning of the ACE cDNA from pig kidney by using the reverse transcriptase-polymerase chain reaction. The complete amino acid sequence deduced from the cDNA contains 1309 residues with a molecular mass of 150 kDa, beginning with a signal peptide of 33 amino acids. Amino acid sequence analysis showed that pig kidney ACE is also probably anchored by a short transmembrane domain located near the C-terminus. This protein contains a tandem duplication of the two homologous amino acid peptidase domain. Each of these two domains bears a putative metal-binding site (His-Glu-Met-Gly-His) identified in mammalian somatic ACE. The alignment of pig ACE amino acid sequence with human, rabbit, and mouse reveals that both two domains have been highly conserved during evolution.

GEOMETRIC NINLINEAR ANALYSIS OF UNERGROUND LAMINATED COMPISITE PIPES (기하학적 비선형을 고려한 지하매설 복합재료 파이프의 해석)

  • 김덕현;이인원;변문주
    • Computational Structural Engineering
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    • v.2 no.1
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    • pp.65-70
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    • 1989
  • An analytical study was conducted using the Galerkin technique to determine behaviour of thin fibrereinforced and laminated composite pipes under soil pressure. Geometric nonlinearity and material linearity have been assumed. It is assumed that vertical and lateral soil pressure are proportional to the depth and lateral displacement of the pipe respectively. It is also assumed that radial shear stress is negligible because the ratio of thickness to the radius of pipe is very small. The above results are verified by the finite element analysis.

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Molecular genetic analysis of phytochelatin synthase genes in Arabidopsis

  • Ha, Suk-Bong
    • Proceedings of the Botanical Society of Korea Conference
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    • 2002.04a
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    • pp.62-72
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    • 2002
  • This study has investigated the biosynthesis and function of the heavy metal binding peptides, the phytochelatins, in plants. PCs are synthesised enzymatically from glutathione by the enzyme PC synthase in the presence of heavy metal ions. Using Arabidopsis thaliana as a model organism cadmium-sensitive, phytochelatin-deficient mutants have been isolated and characterised in previous studies. The cadl mutants have wildtype levels of glutathione, are PC deficient and lack PC synthase activity. Thus, the CADl gene has been proposed to encode PC synthase. The CADl gene was isolated by a positional cloning strategy The gene was mapped and a candidate identified. Each of four cadl mutants had a single base pair change in the candidate gene and the cadmium-sensitive, cadl phenotype was complemented by the candidate gene. This demonstrated the CADl gene had been cloned. A homologous gene in the fission yeast, Schizosaccharomyces pombe was identified through database searches. A targeted-deletion mutation of this gene was constructed and the mutant, like cadl mutants of Arabidopsis, was cadmium-sensitive and PC-deficient. A comparison of the redicted amino acid sequences reveals a highly conserved N-terminal region Presumed to be the catalytic domain and a variable C-terminal region containing multiple Cys residues proposed to be involved in activation of the enzyme by metal ions. Similar genes were also identified in animal species. The Arabidopsis CADl/AtPCSl and S. pombe SpbPCS genes were expressed in E. coli and were shown to be sufficient for glutathione-dependent, heavy metal activate PC synthesis in vitro, thus demonstrating these genes encode PC synthase enzymes. Using RT-PCR, AtPCSl expression appeared to be independent of Cd exposure. However, at higher levels of Cd exposure a AtPCSl-CUS reporter gene construct appeared to be more highly expressed. Using the reporter gene construct, AtPCSl was expressed most tissues. Expression appeared to be greater in younger tissues and same higher levels of expression was observed in some regions, including carpels and the base of siliques. AtPCS2 was a functional gene encoding an active PC synthase. However, its Pattern of expression and the phenotype of a mutant (or antisense line) have not been determined. Assuming the gene is functional then it has clearly been maintained through evolution and must provide some selective advantage. This implies that, at least in some cells or tissue, it is likely to be the dominant PC synthase expressed. This remains to be determined

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A refined Panax ginseng karyotype based on an ultra-high copy 167-bp tandem repeat and ribosomal DNAs

  • Waminal, Nomar Espinosa;Choi, Hong-Il;Kim, Nam-Hoon;Jang, Woojong;Lee, Junki;Park, Jee Young;Kim, Hyun Hee;Yang, Tae-Jin
    • Journal of Ginseng Research
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    • v.41 no.4
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    • pp.469-476
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    • 2017
  • Background: Panax ginseng Meyer (Asian ginseng) has a large nuclear genome size of > 3.5 Gbp in haploid genome equivalent of 24 chromosomes. Tandem repeats (TRs) occupy significant portions of the genome in many plants and are often found in specific genomic loci, making them a valuable molecular cytogenetic tool in discriminating chromosomes. In an effort to understand the P. ginseng genome structure, we characterized an ultrahigh copy 167-bp TR (Pg167TR) and explored its chromosomal distribution as well as its utility for chromosome identification. Methods: Polymerase chain reaction amplicons of Pg167TR were labeled, along with 5S and 45S rDNA amplicons, using a direct nick-translation method. Direct fluorescence in situ hybridization (FISH) was used to analyze the chromosomal distribution of Pg167TR. Results: Recently, we reported a method of karyotyping the 24 chromosome pairs of P. ginseng using rDNA and DAPI (4',6-diamidino-2-phenylindole) bands. Here, a unique distribution of Pg167TR in all 24 P. ginseng chromosomes was observed, allowing easy identification of individual homologous chromosomes. Additionally, direct labeling of 5S and 45S rDNA probes allowed the identification of two additional 5S rDNA loci not previously reported, enabling the refinement of the P. ginseng karyotype. Conclusion: Identification of individual P. ginseng chromosomes was achieved using Pg167TR-FISH. Chromosome identification is important in understanding the P. ginseng genome structure, and our method will be useful for future integration of genetic linkage maps and genome scaffold anchoring. Additionally, it is a good tool for comparative studies with related species in efforts to understand the evolution of P. ginseng.

Phylogenetic study of Korean Geranium(Geraniaceae) based on nrDNA ITS squences (ITS 염기서열에 의한 한국산 쥐손이풀속(Geranium)의 계통학적 연구)

  • Woo, Jeong Hyeon;Park, Seon-Joo
    • Korean Journal of Plant Taxonomy
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    • v.36 no.2
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    • pp.91-108
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    • 2006
  • Phylogenetic analyses were conducted to evaluate evolution and relationship of 16 taxa of Korean Geranium including 3 outgroups using ITS (internal transcribed spacer) squences of nuclear ribosomal DNA. Phylogenetic studies used most parsimony and neighbor-joining methods including bootstrapping and jackknifing analysis. As the result, Korean Geranium forms monophyletic group. In the parsimony tree G. koraiense var. hallasanense situated as the most basal clade and Erianthum group forms one clade by high bootstrap ans jackknife values (100% of bootstrap and jackknife values). G.dahuricum as one of the Krameri group is closely related with Palustre group by very weak relationship (37% of bootstrap and 44% of jackknife values) and the node collapse in the strict tree. G. Knuthii which was one of wilfordii group is closely related with Koreanum group. G. sibiricum, one of Sibiricum group, is the most closest relationship with G. soboliferum and these species are sister to G. krameri. G. tripartitum and G. wilfordii which are wilfordii group are linked to G. nepalense, G. thunbergii f. pallidum and G. thunbergii. This result suggested that the phylogenetic analysis of ITS sequences should be useful to address phylogenetic questions on the genus Korean Geranium.

FOLLOW-UP OBSERVATIONS TOWARD PLANCK COLD CLUMPS WITH GROUND-BASED RADIO TELESCOPES

  • LIU, TIE;WU, YUEFANG;MARDONES, DIEGO;KIM, KEE-TAE;MENTEN, KARL M.;TATEMATSU, KEN;CUNNINGHAM, MARIA;JUVELA, MIKA;ZHANG, QIZHOU;GOLDSMITH, PAUL F;LIU, SHENG-YUAN;ZHANG, HUA-WEI;MENG, FANYI;LI, DI;LO, NADIA;GUAN, XIN;YUAN, JINGHUA;BELLOCHE, ARNAUD;HENKEL, CHRISTIAN;WYROWSKI, FRIEDRICH;GARAY, GUIDO;RISTORCELLI, ISABELLE;LEE, JEONG-EUN;WANG, KE;BRONFMAN, LEONARDO;TOTH, L. VIKTOR;SCHNEE, SCOTT;QIN, SHENGLI;AKHTER, SHAILA
    • Publications of The Korean Astronomical Society
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    • v.30 no.2
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    • pp.79-82
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    • 2015
  • The physical and chemical properties of prestellar cores, especially massive ones, are still far from being well understood due to the lack of a large sample. The low dust temperature (< 14 K) of Planck cold clumps makes them promising candidates for prestellar objects or for sources at the very initial stages of protostellar collapse. We have been conducting a series of observations toward Planck cold clumps (PCCs) with ground-based radio telescopes. In general, when compared with other star forming samples (e.g. infrared dark clouds), PCCs are more quiescent, suggesting that most of them may be in the earliest phase of star formation. However, some PCCs are associated with protostars and molecular outflows, indicating that not all PCCs are in a prestellar phase. We have identified hundreds of starless dense clumps from a mapping survey with the Purple Mountain Observatory (PMO) 13.7-m telescope. Follow-up observations suggest that these dense clumps are ideal targets to search for prestellar objects.

Alu-Derived Alternative Splicing Events Specific to Macaca Lineages in CTSF Gene

  • Lee, Ja-Rang;Park, Sang-Je;Kim, Young-Hyun;Choe, Se-Hee;Cho, Hyeon-Mu;Lee, Sang-Rae;Kim, Sun-Uk;Kim, Ji-Su;Sim, Bo-Woong;Song, Bong-Seok;Jeong, Kang-Jin;Lee, Youngjeon;Jin, Yeung Bae;Kang, Philyong;Huh, Jae-Won;Chan, Kyu-Tae
    • Molecules and Cells
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    • v.40 no.2
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    • pp.100-108
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    • 2017
  • Cathepsin F, which is encoded by CTSF, is a cysteine proteinase ubiquitously expressed in several tissues. In a previous study, novel transcripts of the CTSF gene were identified in the crab-eating monkey deriving from the integration of an Alu element-AluYRa1. The occurrence of AluYRa1-derived alternative transcripts and the mechanism of exonization events in the CTSF gene of human, rhesus monkey, and crabeating monkey were investigated using PCR and reverse transcription PCR on the genomic DNA and cDNA isolated from several tissues. Results demonstrated that AluYRa1 was only integrated into the genome of Macaca species and this lineage-specific integration led to exonization events by producing a conserved 3' splice site. Six transcript variants (V1-V6) were generated by alternative splicing (AS) events, including intron retention and alternative 5' splice sites in the 5' and 3' flanking regions of CTSF_AluYRa1. Among them, V3-V5 transcripts were ubiquitously expressed in all tissues of rhesus monkey and crab-eating monkey, whereas AluYRa1-exonized V1 was dominantly expressed in the testis of the crab-eating monkey, and V2 was only expressed in the testis of the two monkeys. These five transcript variants also had different amino acid sequences in the C-terminal region of CTSF, as compared to reference sequences. Thus, species-specific Alu-derived exonization by lineage-specific integration of Alu elements and AS events seems to have played an important role during primate evolution by producing transcript variants and gene diversification.

Function and Molecular Ecology Significance of Two Catechol-Degrading Gene Clusters in Pseudomonas putida ND6

  • Shi, Sanyuan;Yang, Liu;Yang, Chen;Li, Shanshan;Zhao, Hong;Ren, Lu;Wang, Xiaokang;Lu, Fuping;Li, Ying;Zhao, Huabing
    • Journal of Microbiology and Biotechnology
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    • v.31 no.2
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    • pp.259-271
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    • 2021
  • Many bacteria metabolize aromatic compounds via catechol as a catabolic intermediate, and possess multiple genes or clusters encoding catechol-cleavage enzymes. The presence of multiple isozyme-encoding genes is a widespread phenomenon that seems to give the carrying strains a selective advantage in the natural environment over those with only a single copy. In the naphthalene-degrading strain Pseudomonas putida ND6, catechol can be converted into intermediates of the tricarboxylic acid cycle via either the ortho- or meta-cleavage pathways. In this study, we demonstrated that the catechol ortho-cleavage pathway genes (catBICIAI and catBIICIIAII) on the chromosome play an important role. The catI and catII operons are co-transcribed, whereas catAI and catAII are under independent transcriptional regulation. We examined the binding of regulatory proteins to promoters. In the presence of cis-cis-muconate, a well-studied inducer of the cat gene cluster, CatRI and CatRII occupy an additional downstream site, designated as the activation binding site. Notably, CatRI binds to both the catI and catII promoters with high affinity, while CatRII binds weakly. This is likely caused by a T to G mutation in the G/T-N11-A motif. Specifically, we found that CatRI and CatRII regulate catBICIAI and catBIICIIAII in a cooperative manner, which provides new insights into naphthalene degradation.