• Asian-Australasian Journal of Animal Sciences
• /
• v.25 no.6
• /
• pp.789-793
• /
• 2012

Fertilization of the oocyte commences embryogenesis during which maternally inherited mRNAs are degraded and the embryonic genome is activated. Transcription of embryonic mRNA is initiated by embryonic genome activation (EGA). RNA polymerase II (RNA Pol II) is responsible for the synthesis of mRNAs and most small nuclear RNAs, and consists of 12 subunits, the largest of which characteristically harbors a unique C-terminal domain (CTD). Transcriptional activity of RNA Pol II is highly regulated, in particular, by phosphorylation of serine residues in the CTD. Here, we have shown the presence of RNA Pol II CTD phosphoisoforms in porcine oocytes and preimplantation embryos. The distribution pattern as well as phosphorylation dynamics in germinal vesicles and during embryogenesis differed in developmental stages with these isoforms, indicating a role of RNA Pol II CTD phosphorylation at the serine residue in transcriptional activation during both oocyte growth and embryonic genome activation. We additionally examined the effects of the RNA Pol II inhibitor, ${\alpha}$-amanitin, on embryo development. Our results show that inhibition of polymerase, even at very early stages and for a short period of time, dramatically impaired blastocyst formation. These findings collectively suggest that the functionality of maternal RNA Pol II, and consequently, expression of early genes regulated by this enzyme are essential for proper embryo development.

• Journal of Microbiology and Biotechnology
• /
• v.26 no.12
• /
• pp.2087-2097
• /
• 2016

Most cold-adapted enzymes possess higher $K_m$ and $k_{cat}$ values than those of their mesophilic counterparts to maximize the reaction rate. This characteristic is often ascribed to a high structural flexibility and improved dynamics in the active site. However, this may be less convincing to cold-adapted metabolic enzymes, which work at substrate concentrations near $K_m$. In this respect, cold adaptation of a shikimate kinase (SK) in the shikimate pathway from psychrophilic Colwellia psychrerythraea (CpSK) was characterized by comparing it with a mesophilic Escherichia coli homolog (EcSK). The optimum temperatures for CpSK and EcSK activity were approximately $30^{\circ}C$ and $40^{\circ}C$, respectively. The melting points were $33^{\circ}C$ and $45^{\circ}C$ for CpSK and EcSK, respectively. The ${\Delta}G_{H_2O}$ (denaturation in the absence of denaturing agent) values were 3.94 and 5.74 kcal/mol for CpSK and EcSK, respectively. These results indicated that CpSK was a cold-adapted enzyme. However, contrary to typical kinetic data, CpSK had a lower $K_m$ for its substrate shikimate than most mesophilic SKs, and the $k_{cat}$ was not increased. This observation suggested that CpSK may have evolved to exhibit increased substrate affinity at low intracellular concentrations of shikimate in the cold environment. Sequence analysis and homology modeling also showed that some important salt bridges were lost in CpSK, and higher Arg residues around critical Arg 140 seemed to increase flexibility for catalysis. Taken together, these data demonstrate that CpSK exhibits characteristics of cold adaptation with unusual kinetic parameters, which may provide important insights into the cold adaptation of metabolic enzymes.

• Mass Spectrometry Letters
• /
• v.3 no.4
• /
• pp.93-100
• /
• 2012

Cellular processes such as proliferation, differentiation, and adaptation to environmental changes are regulated by protein phosphorylation. In order to enhance the understanding of molecular dynamics for biological process in detail, it is necessary to develop sensitive and comprehensive analytical methods for the determination of protein phosphorylation. Neural stem cells hold great promise for neural repair following an injury or disease. In this study, we made differentiated oligodendrocytes from human neural stem cells using over-expression of olig2 gene. We confirmed using quantitative phosphoproteome analysis approach that combines stable isotope labeling by amino acids in cell culture (SILAC) and $TiO_2$ micro-column for phosphopeptide enrichment with $MS^2$ and $MS^3$ mass spectrometry. We detected 275 phosphopeptides which were modulated at least 2-fold between human neural stem cells and oligodendrocytes. Among them, 23 phosphoproteins were up-regulated in oligodendrocytes and 79 phosphoproteins were up-regulated in F3 cells.

• Genomics & Informatics
• /
• v.10 no.3
• /
• pp.145-152
• /
• 2012

Chromatin structure and dynamics that are influenced by epigenetic marks, such as histone modification and DNA methylation, play a crucial role in modulating gene transcription. To understand the relationship between histone modifications and regulatory elements in breast cancer cells, we compared our chromatin immunoprecipitation sequencing (ChIP-Seq) histone modification patterns for histone H3K4me1, H3K4me3, H3K9/16ac, and H3K27me3 in MCF-7 cells with publicly available formaldehyde-assisted isolation of regulatory elements (FAIRE)-chip signals in human chromosomes 8, 11, and 12, identified by a method called FAIRE. Active regulatory elements defined by FAIRE were highly associated with active histone modifications, like H3K4me3 and H3K9/16ac, especially near transcription start sites. The H3K9/16ac-enriched genes that overlapped with FAIRE signals (FAIRE-H3K9/14ac) were moderately correlated with gene expression levels. We also identified functional sequence motifs at H3K4me1-enriched FAIRE sites upstream of putative promoters, suggesting that regulatory elements could be associated with H3K4me1 to be regarded as distal regulatory elements. Our results might provide an insight into epigenetic regulatory mechanisms explaining the association of histone modifications with open chromatin structure in breast cancer cells.

• Animal cells and systems
• /
• v.5 no.3
• /
• pp.253-262
• /
• 2001

Ligand-receptor clustering event is the most important step in leukocyte adhesion and spreading on endothelial cells. Intercellular adhesion molecule-1 (ICAM-1) has been shown to enhance leukocyte adhesion, but the molecular event during the process of adhesion is unclear. To visualize the dynamics of ICAM-1 movement during adhesion, we have engineered stable Chinese hamster ovary cell lines expressing ICAM-1 fused to a green fluorescent protein (IC1_GFP/CHO) and examined them under the fluorescence microscopy. The transfection of IC1_GFP alone in these cells was sufficient to support the adhesion of K562 cells that express $\alpha$L$\beta$2 (LFA-1) integrin stimulated by CBR LFA-1/2 mAb. This phenomenon was mediated by ICAM-1-LFA-1 interactions, as an mAb that specifically inhibits ICAM-1-LFA-1 interaction (RRl/l) completely abolished the adhesion of LFA-1＊ cells to IC1_ GFP/CHO cells. We found that the characteristic of adhesion was followed almost immediately (~10 min) by the rapid accumulation of ICAM-1 on CHO cells at a tight interface between the two cells. Interestingly, ICI_GFP/CHO cells projected plasma membrane and encircled approximately half surface of LFA-1+ cells, as defined by confocal microscopy. This unusual phenomenon was also confirmed on HUVEC after adhesion of LFA-1＊ cells. Neither cytochalasin D nor 2,3-butanedione 2-monoxime an inhibitor of myosin light chain kinase blocked LFA-1-mediated ICAM-1 clustering, indicating that actin cytoskeleton and myosin-dependent contractility are not necessary for ICAM-1 clustering. Taken together, we suggest that leukocyte adhesion to endothelial cells induces specialized form of ICAM-1 clustering that is distinct from immunological synapse mediated by T cell interaction with antigen presenting cells.

• The Journal of the Convergence on Culture Technology
• /
• v.6 no.1
• /
• pp.23-29
• /
• 2020

Since the movements of a machine are driven by a certain principle, there is a fundamental principle that governs the movement of the machine, no matter how complex and diverse it may be. Autonomous movements are a way to avoid repetition, to approach more humane and more natural factors. That's all the more so when it comes to original movements, not cloning. So now, the dynamics of the machine are more multi-sensory by human participation. It is challenging the potential to be expressed and is gradually expanding its sphere of expression in the boundaries of fact and cloning. Theo Jansen's works have changed from period to period to adapt to different circumstances, indicating that his decades-long work has continued to evolve from early to present. The evolution's focus, among other things, can be seen as having the same source of circular motion and horizontal reciprocating motion, which are repeated from the principle of organic coupling between the parts that make up the object.

• Journal of Science of Art and Design
• /
• v.4
• /
• pp.91-122
• /
• 2002

`Bio morphism` are constituted in paintings where the artists try to embody the elementary properties of living creature as of growth and durability. They are the most appropriate concept of painting to harmonize human being with nature closely. The formative ways of them attach great importance to both unconsciousness and desire , as well as variations or dynamics, by noticing a flow of natural senses and feelings of human being. In other words, the formative ways are based on a recognition of nature as the intrinsic force of life, with the result that aesthetics of incompleteness is embodied in images. Therefore they are clearly distinguished from that of functional, geometric images. A tendency of painting at that time, in a word, 'return to figure and expression', means a conversion into organic images like the incomplete, atypical, and biomorphic forms, while denying the mechanical or geometric. Elizabeth Murray are analyzed, for these works are remarkable in the characteristics of 'Bio morphism'. Consequently the features of organic images, that is, 'the formative acceptance of natural figures, or an informality' and 'the force of free will, or an incompleteness', could obviously be revealed. It is a type that obtains a motif out of natural figures like an animal, a plant, or the concrete figures of human being. In conclusion, this thesis is focused on not only emphasizing that 'Bio morphism' were a major tendency among the various trends of postmodern painting in the 20th century, but also analysing both the painterly formation of organic images and the structure of them. In addition to these points, it is a central aim to evoke that Bio morphism should accurately be evaluated and positioned in postmodern painting. A new recognition of 'Bio morphism' is a peculiarity of the times that reflects a cultural aspect of the present, hence it should be recognized as another way to approach the postmodern painting.

• Biomolecules & Therapeutics
• /
• v.27 no.6
• /
• pp.530-539
• /
• 2019

Brain aging is an inevitable process characterized by structural and functional changes and is a major risk factor for neurodegenerative diseases. Most brain aging studies are focused on neurons and less on astrocytes which are the most abundant cells in the brain known to be in charge of various functions including the maintenance of brain physical formation, ion homeostasis, and secretion of various extracellular matrix proteins. Altered mitochondrial dynamics, defective mitophagy or mitochondrial damages are causative factors of mitochondrial dysfunction, which is linked to age-related disorders. Etoposide is an anti-cancer reagent which can induce DNA stress and cellular senescence of cancer cell lines. In this study, we investigated whether etoposide induces senescence and functional alterations in cultured rat astrocytes. Senescence-associated ${\beta}$-galactosidase (SA-${\beta}$-gal) activity was used as a cellular senescence marker. The results indicated that etoposide-treated astrocytes showed cellular senescence phenotypes including increased SA-${\beta}$-gal-positive cells number, increased nuclear size and increased senescence-associated secretory phenotypes (SASP) such as IL-6. We also observed a decreased expression of cell cycle markers, including PhosphoHistone H3/Histone H3 and CDK2, and dysregulation of cellular functions based on wound-healing, neuronal protection, and phagocytosis assays. Finally, mitochondrial dysfunction was noted through the determination of mitochondrial membrane potential using tetramethylrhodamine methyl ester (TMRM) and the measurement of mitochondrial oxygen consumption rate (OCR). These data suggest that etoposide can induce cellular senescence and mitochondrial dysfunction in astrocytes which may have implications in brain aging and neurodegenerative conditions.

• Molecules and Cells
• /
• v.44 no.2
• /
• pp.79-87
• /
• 2021

Chromatin dynamics is essential for maintaining genomic integrity and regulating gene expression. Conserved bromodomain-containing AAA+ ATPases play important roles in nucleosome organization as histone chaperones. Recently, the high-resolution cryo-electron microscopy structures of Schizosaccharomyces pombe Abo1 revealed that it forms a hexameric ring and undergoes a conformational change upon ATP hydrolysis. In addition, single-molecule imaging demonstrated that Abo1 loads H3-H4 histones onto DNA in an ATP hydrolysis-dependent manner. However, the molecular mechanism by which Abo1 loads histones remains unknown. Here, we investigated the details concerning Abo1-mediated histone loading onto DNA and the Abo1-DNA interaction using single-molecule imaging techniques and biochemical assays. We show that Abo1 does not load H2A-H2B histones. Interestingly, Abo1 deposits multiple copies of H3-H4 histones as the DNA length increases and requires at least 80 bp DNA. Unexpectedly, Abo1 weakly binds DNA regardless of ATP, and neither histone nor DNA stimulates the ATP hydrolysis activity of Abo1. Based on our results, we propose an allosteric communication model in which the ATP hydrolysis of Abo1 changes the configuration of histones to facilitate their deposition onto DNA.

• Journal of Astronomy and Space Sciences
• /
• v.39 no.2
• /
• pp.67-77
• /
• 2022

The elements that impact the dynamics and collaborations of waves and particles in the magnetosphere of planets have been considered here. Saturn's internal magnetosphere is determined by substantiated instabilities and discovered to be an exceptional zone of wave activity. Interchanged instability is found to be one of the responsible events in view of temperature anisotropy and energization processes of magnetospheric species. The generated active ions alongside electrons that constitute the populations of highly magnetized planets like Saturn's ring electron current are taken into consideration in the current framework. The previous and similar method of characteristics and the perturbed distribution function have been used to derive dispersion relation. In incorporating this investigation, the characteristics of electromagnetic ion cyclotron wave (EMIC) waves are determined by the composition of ions in plasmas through which the waves propagate. The effect of ring distribution illustrates non-monotonous description on growth rate (GR) depending upon plasma parameters picked out. Observations made by Cassini found appropriate for modern study, have been applied to the Kronian magnetosphere. Using Maxwellian ring distribution function of ions and detailed mathematical formulation, an expression for dispersion relation as well as GR and real frequency (RF) are evaluated. Analysis of plasma parameters shows that, proliferating EMIC waves are not developed much when propagation is parallelly aligned with magnetosphere as compared to waves propagating in oblique direction. GR for the oblique case, is influenced by temperature anisotropy as well as by alternating current (AC) frequency, whereas it is much affected only by AC frequency for parallel propagating waves.