• Microbiology and Biotechnology Letters
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• v.19 no.1
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• pp.37-44
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• 1991

- I aimed to isolate trans-acting factors involved in the basal expression level of eukaryotic genes. One of the yeast histidine biosynthetic gene, HIS5 was taken as a model for this study. HIS5 gene has a substantial basal level in amino acid rich medium and is derepressed if starved for any single amino acid. The derepression is mediated by cis-acting DNA sequences 5'-TGACTC-3' found in 5' non-transcribed region of the gene and trans-acting factors including GCN4 as positive factor and its negative factor GCDI 7, and GCNZ as a negative factor of GCD17. I first investigated the role of these trans-acting factors in HIS5 basal expression level by using HIS5-pH05 fusion in which expression of pH05 gene encoding inorganic phosphate-repressible acid phosphatase (APase) is regulated by HIS5 promoter. Strain with gcn2 or gcn4 mutation showed 3 to 4 fold lower APase activity than wild type. The level of APase activity was similar in gcn2 and gcn4 mutants. Trans-acting factors involved in basal level were identified by isolating 14 mutants showing increased expression of HISSPH05 fusion from gcn4 background. All the mutants carry a single nuclear recessive mutation and fall into four complementation groups, designated as bell (basal expression level), be12, be23 and be14.

• Proceedings of the Microbiological Society of Korea Conference
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• 2002.10a
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• pp.135-139
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• 2002

Yeast cells respond to condition of amino acid starvation by synthesizing GCN4, a typical eukaryotic transcriptional activator, which regulates the expression of many amino acids biosynthetic genes. By introducing point mutations in the DNA binding domain of GCN4, mutants with normal DNA binding activity but defective in transcriptional activity were isolated to identify unknown proteins that could suppress the mutant phenotype under an amino acid depletion condition. As a result, SSB(Stress-Seventy B) subfamily proteins were identified as suppressors of mutant GCN4. SSB proteins were known as a member of yeast hsp70 family that probably aids passage of nascent chain through ribosomes. Among them, the mechanism of suppression by SSB2 on the defective GCN4 mutant strains is under investigation. Gcn4p directly interacts with Ssb2p through the basic DNA binding domain of GCN4. It suggests the possibility that physical interaction might induce the transcriptional activation of Gcn4p.

• Molecules and Cells
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• v.24 no.2
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• pp.194-199
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• 2007

GCN4 is a typical eukaryotic transcriptional activator that is implicated in the expression of many genes involved in amino acids and purine biosyntheses under stress conditions. It is degraded by 26S proteasomes following ubiquitination. However, the immediate receptor for ubiquitinated Gcn4p has not yet been identified. We investigated whether ubiquitinated Gcn4p binds directly to Rpn10p as the ubiquitinated substrate receptor of the 26S proteasome. We found that the level of Gcn4p increased in cells deleted for Rpn10p but not in cells deleted for RAD23 and DSK2, the other ubiquitinated substrate receptors and, unlike Rpn10p, neither of these proteins recognized ubiquitinated Gcn4p. These results suggest that Rpn10p is the receptor that binds the polyubiquitin chain during ubiquitin-dependent proteolysis of Gcn4p.

• Journal of Microbiology and Biotechnology
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• v.9 no.1
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• pp.122-125
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• 1999

Gcn4p, a transcriptional activator protein of the yeast, Sacchromyces cerevisiae, binds to the specific sequence in the promoters of many amino acid biosynthetic genes for general control. The serine residue (Ser 242) of Gcn4p directly contacts the DNA. Here, for inspecting the DNA binding properties and the level of transcriptional activation of Gcn4p, we introduced a polymerase chain reaction (PCR) site-directed saturation mutation library into the Ser 242 site using 2 outside primers and 2 oligonucleotides with its codons fully degenerated. The sequencing analysis of 146 samples revealed the even nucleotide distribution within the experimental error showing 23, 26, 25, and 26％ frequency of U, C, A, and G bases, respectively. This method turned out to be a simple, fast, and economical method for constructing a library of all 20 amino acids at specific codon.

• Biomedical Science Letters
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• v.18 no.2
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• pp.165-168
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• 2012

Hox proteins containing DNA-binding homedomain act as transcription factors important for anteroposterior body patterning during vertebrate embryogenesis. However, the precise mechanisms by which signal pathways are transduced to regulate the Hox gene expression are not clear. In the course of an attempt to isolate an upstream regulatory factor(s) controlling Hox genes, protein kinase B alpha (Akt1) has been identified as a putative regulator of Hox genes through in silico analysis (GEO profile). In the Gene Expression Omnibus (GEO) dataset GDS1784 at the NCBI (National Center for Biotechnology Information) site, Hox genes were differentially expressed depending on the presence or absence of Akt1. Since it was not well known how Akt1 regulates the specific Hox genes, whose transcription was reported to be regulated by epigenetic modifications such as histone acetylation, methylation etc., the expression of Gcn5, a histone acetyltransferase (HAT), was analyzed in wild type (WT) as well as in $Akt1^{-/-}$ mouse embryonic fibroblast (MEF) cells. RT-PCR analysis revealed that the amount of Gcn5 mRNA was similar in both WT and $Akt1^{-/-}$ MEFs. However, the protein level of Gcn5 was significantly increased in $Akt1^{-/-}$ MEF cells. The half life of Gcn5 was 1 hour in wild type whereas 8 hours in $Akt1^{-/-}$ MEF. These data all together, indicate that Gcn5 is post-transcriptionally down-regulated and the protein stability is negatively regulated by Akt1 in MEF cells.

• The Journal of Korea Institute of Information, Electronics, and Communication Technology
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• v.14 no.4
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• pp.314-322
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• 2021

Skeleton-based action recognition has attracted considerable attention in human action recognition. Recent methods for skeleton-based action recognition employ spatiotemporal graph convolutional networks (GCNs) and have remarkable performance. However, most of them have heavy computational complexity for robust action recognition. To solve this problem, we propose a shuffle graph convolutional network (SGCN) which is a lightweight graph convolutional network using pointwise group convolution rather than pointwise convolution to reduce computational cost. Our SGCN is composed of spatial and temporal GCN. The spatial shuffle GCN contains pointwise group convolution and part shuffle module which enhances local and global information between correlated joints. In addition, the temporal shuffle GCN contains depthwise convolution to maintain a large receptive field. Our model achieves comparable performance with lowest computational cost and exceeds the performance of baseline at 0.3% and 1.2% on NTU RGB+D and NTU RGB+D 120 datasets, respectively.

• KSII Transactions on Internet and Information Systems (TIIS)
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• v.14 no.5
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• pp.1909-1928
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• 2020

Predicting human mobility has always been an important task in Location-based Social Network. Previous efforts fail to capture spatial dependence effectively, mainly reflected in weakening the location topology information. In this paper, we propose a neural network-based method which can capture spatial-temporal dependence to predict the next location of a person. Specifically, we involve a graph convolutional network (GCN) based on a seq2seq framework to capture the location topology information and temporal dependence, respectively. The encoder of the seq2seq framework first generates the hidden state and cell state of the historical trajectories. The GCN is then used to generate graph embeddings of the location topology graph. Finally, we predict future trajectories by aggregated temporal dependence and graph embeddings in the decoder. For evaluation, we leverage two real-world datasets, Foursquare and Gowalla. The experimental results demonstrate that our model has a better performance than the compared models.

• Journal of Korea Multimedia Society
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• v.25 no.11
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• pp.1643-1652
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• 2022

Most of the existing machine reading research has used Recurrent Neural Network (RNN) and Convolutional Neural Network (CNN) algorithms as networks. Among them, RNN was slow in training, and Question Answering Network (QANet) was announced to improve training speed. QANet is a model composed of CNN and self-attention. CNN extracts semantic and syntactic information well from the local corpus, but there is a limit to extracting the corresponding information from the global corpus. Graph Convolutional Networks (GCN) extracts semantic and syntactic information relatively well from the global corpus. In this paper, to take advantage of this strength of GCN, we propose I-QANet, which changed the CNN of QANet to GCN. The proposed model performed 1.2 times faster than the baseline in the Stanford Question Answering Dataset (SQuAD) dataset and showed 0.2% higher performance in Exact Match (EM) and 0.7% higher in F1. Furthermore, in the Korean Question Answering Dataset (KorQuAD) dataset consisting only of Korean, the learning time was 1.1 times faster than the baseline, and the EM and F1 performance were also 0.9% and 0.7% higher, respectively.

• Molecules and Cells
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• v.38 no.12
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• pp.1037-1043
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• 2015

The TAZ activator 2-butyl-5-methyl-6-(pyridine-3-yl)-3-[2'-(1H-tetrazole-5-yl)-biphenyl-4-ylmethyl]-3H-imidazo[4,5-b]pyridine] (TM-25659) inhibits adipocyte differentiation by interacting with peroxisome proliferator-activated receptor gamma. 1 TM-25659 was previously shown to decrease weight gain in a high fat (HF) diet-induced obesity (DIO) mouse model. However, the fundamental mechanisms underlying the effects of TM-25659 remain unknown. Therefore, we investigated the effects of TM-25659 on skeletal muscle functions in C2 myotubes and C57BL/6J mice. We studied the molecular mechanisms underlying the contribution of TM-25659 to palmitate (PA)-induced insulin resistance in C2 myotubes. TM-25659 improved PA-induced insulin resistance and inflammation in C2 myotubes. In addition, TM-25659 increased FGF21 mRNA expression, protein levels, and FGF21 secretion in C2 myotubes via activation of GCN2 pathways (GCN2-$phosphoelF2{\alpha}$-ATF4 and FGF21). This beneficial effect of TM-25659 was diminished by FGF21 siRNA. C57BL/6J mice were fed a HF diet for 30 weeks. The HF-diet group was randomly divided into two groups for the next 14 days: the HF-diet and HF-diet + TM-25659 groups. The HF diet + TM-25659-treated mice showed improvements in their fasting blood glucose levels, insulin sensitivity, insulin-stimulated Akt phosphorylation, and inflammation, but neither body weight nor food intake was affected. The HF diet + TM-25659-treated mice also exhibited increased expression of both FGF21 mRNA and protein. These data indicate that TM-25659 may be beneficial for treating insulin resistance by inducing FGF21 in models of PA-induced insulin resistance and HF diet-induced insulin resistance.

• BMB Reports
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• v.52 no.3
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• pp.163-164
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• 2019

The ribosomal synthesis of proteins in the eukaryotic cytosol has always been thought to start from the unformylated N-terminal (Nt) methionine (Met). In contrast, in virtually all nascent proteins in bacteria and eukaryotic organelles, such as mitochondria and chloroplasts, Nt-formyl-methionine (fMet) is the first building block of ribosomal synthesis. Through extensive approaches, including mass spectrometric analyses of the N-termini of proteins and molecular genetic techniques with an affinity-purified antibody for Nt-formylation, we investigated whether Nt-formylated proteins could also be produced and have their own metabolic fate in the cytosol of a eukaryote, such as yeast Saccharomyces cerevisiae. We discovered that Nt-formylated proteins could be generated in the cytosol by yeast mitochondrial formyltransferase (Fmt1). These Nt-formylated proteins were massively upregulated in the stationary phase or upon starvation for specific amino acids and were crucial for the adaptation to specific stresses. The stress-activated kinase Gcn2 was strictly required for the upregulation of Nt-formylated proteins by regulating the activity of Fmt1 and its retention in the cytosol. We also found that the Nt-fMet residues of Nt-formylated proteins could be distinct N-terminal degradation signals, termed fMet/N-degrons, and that Psh1 E3 ubiquitin ligase mediated the selective destruction of Nt-formylated proteins as the recognition component of a novel eukaryotic fMet/N-end rule pathway, termed fMet/N-recognin.