• Journal of Plant Biotechnology
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• v.45 no.4
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• pp.364-368
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• 2018

The R2R3-type protein IbMYB1 transcription factor is a key regulator for anthocyanin biosynthesis in the storage roots of sweet potatoes. It was previously demonstrated that the IbMYB1 expression stimulates anthocyanin pigmentation in tobacco leaves, arabidopsis and storage roots of sweet potatoes. In this study, we generated the transgenic potato plants that express the IbMYB1 genes, which accumulated high levels of anthocyanins under the control of either the tuber-specific patatin (PAT) promoter or oxidative stress-inducible peroxidase anionic 2 (SWPA2) promoter. The PAT-MYB1 transgenic lines exhibited higher anthocyanin levels in the tuber than the empty vector control (EV) or SWPA2-MYB1 plants. When combined, our results indicated that overexpression of the IbMYB1 is a highly promising strategy for the generation of transgenic plants with enhanced tissue specific anthocyanin production.

• Molecules and Cells
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• v.43 no.11
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• pp.945-952
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• 2020

Hypoxia induces the expression of several genes through the activation of a master transcription factor, hypoxia-inducible factor (HIF)-1α. This study shows that hypoxia strongly induced the expression of two carboxypeptidases (CP), CPA4 and CPE, in an HIF-1α-dependent manner. The hypoxic induction of CPA4 and CPE gene was accompanied by the recruitment of HIF-1α and upregulation in the active histone modification, H3K4me3, at their promoter regions. The hypoxic responsiveness of CPA4 and CPE genes was observed in human adipocytes, human adipose-derived stem cells, and human primary fibroblasts but not mouse primary adipocyte progenitor cells. CPA4 and CPE have been identified as secreted exopeptidases that degrade and process other secreted proteins and matrix proteins. This finding suggests that hypoxia changes the microenvironment of the obese hypoxic adipose tissue by inducing the expression of not only adipokines but also peptidases such as CPA4 and CPE.

• Journal of Animal Reproduction and Biotechnology
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• v.39 no.2
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• pp.81-87
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• 2024

Background: Platelet-derived growth factor receptor alpha (PDGFRα) is essential for various biological processes, including fetal Leydig cell differentiation. The PDGFRα^EGFP mouse model, which expresses an eGFP fusion gene under the native Pdgfrα promoter, serves as a valuable resource for exploring PDGFRα's expression and function in vivo. This study investigates PDGFRα expression in adult testicular cells using PDGFRα^EGFP mouse model. Methods: Genotyping PCR and gel electrophoresis were used to confirm the zygosity of PDGFRα^EGFP mice. Histological examination and fluorescence imaging were used to identify PDGFRα expression within testicular tissue. Immunohistochemical analysis assessed the co-expression of PDGFRα with c-Kit, ANO-1, and TASK-1 in testicular cells. Results: Genotyping confirmed the heterozygous status of the mice, which is crucial for studies due to the embryonic lethal phenotype observed in homozygotes. Histological and fluorescence imaging revealed that PDGFRα⁺ cells were primarily located in the interstitial spaces of the testis, specifically within Leydig cells and peritubular myoid cells (PMCs). Immunohistochemical results showed PDGFRα co-localization with c-Kit and ANO-1 in Leydig cells and a complete co-localization with TASK-1 in both Leydig cells and PMCs. Conclusions: The findings demonstrate specific expression of PDGFRα in Leydig cells and PMCs in adult testicular tissue. The co-expression of PDGFRα with c-Kit, ANO-1, and TASK-1 suggests complex regulatory mechanisms, possibly influencing testicular function and broader physiological processes.

• Natural Product Sciences
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• v.1 no.1
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• pp.31-42
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• 1995

One hundred plants in 43 families used in Malaysian traditional medicine were screened for tumour promoting activity using two methods, the technique of activation of latent Epstein-Barr Virus (EBV) in Raji cells and the irritation test on mouse ear. Extracts of only eight plants belonging to the Euphorbiaceae were found to possess EBV activation factor and to give a positive irritation test in mouse ear. These plants included Euphorbia tirucalli L., E. splendes, Jatropha podagrica, J. gossypyfolia L., Pedilanihus tithymaloides (L.) Poitt., Croton argyratus Bl., Exocoecaria agallocha L. and Codiaeum variegatum (L.) Bl. Seven of these plants are used internally in Malaysian traditional medicine. As such, they pose potential danger in the promotion of initiated cells of the mucosal tissue towards disease. Further studies are required to assess the epidemiological impact of these plants in the development of disease.

• Journal of Microbiology
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• v.39 no.3
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• pp.186-194
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• 2001

Scrub typhus, caused by Orientia tsutsugamushi infection, is clinically and histopathologically characterized by local as well as systemic inflammatory reactions, indicating that orientiae induce mechanisms that amplify the inflammatory response. To reveal underlying mechanisms of chemoattraction and activation of responding leukocytes, expression of chemokine and tumor necrosis factor alpha (TNF-$\alpha$) genes in murine peritoneal macrophages after infection with the obligate intracellular bacterium Ο.tsutsugamushi was investigated. The genes that were unregulated included macrophage inflammatory proteins l$\alpha$/$\beta$(MIP-l$\alpha$/$\beta$), MIP-2, monocyte chemoattractant protein 1(MCP-1), RANTES (regulated upon activation, normal T-cell expressed and secreted), gamma-interferon-inducible protein 10(IP-10) and TNF-$\alpha$. Peak expression of these chemokines and TNF-$\alpha$ was observed between 1 and 3 h after infection. These responses returned to or approached baseline preinfection levels 6 h after challenge. Semiquantitative reverse transcription (RT)-PCR analysis revealed dramatic Increases during infection in the steady-state levels of mRNA ceding for the inhibitory subunit of NF-kB (IkB$\alpha$), whose transcription is enhanced by binding of NF-kB within the IkB$\alpha$promoter region. Thus, Ο. tsutsugamushi appears to be a stung inducer of chemokines and TNF-$\alpha$ which may significantly contribute to inflammation and tissue damage observed in scrub typhus by attracting and activating phagocytic leukocytes.

• Asian Pacific Journal of Cancer Prevention
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• v.14 no.11
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• pp.6803-6806
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• 2013

Background: Interferon regulatory factor 6 (IRF6) is a transcription factor with distinct and conserved DNA and protein binding domains. Mutations within the protein binding domain have been significantly observed in subjects with orofacial cleft relative to healthy controls. In addition, recent studies have identified loss of expression of IRF6 due to promoter hypermethylation in cutaneous squamous cell carcinomas. Since mutational events occurring within the conserved domains are likely to affect the function of a protein, we investigated whether regions within the IRF6 gene that encodes for the conserved protein binding domain carried mutations in oral squamous cell carcinoma (OSCC). Materials and Methods: Total chromosomal DNA extracted from 32 post surgical OSCC tissue samples were amplified using intronic primers flanking the exon 7 of IRF6 gene, which encodes for the major region of protein binding domain. The PCR amplicons from all the samples were subsequently resolved in a 1.2% agarose gel, purified and subjected to direct sequencing to screen for mutations. Results: Sequencing analysis resulted in the identification of a mutation within exon 7 of IRF6 that occurred in heterozygous condition in 9% (3/32) of OSCC samples. The wild type codon TTC at position 252 coding for phenylalanine was found to be mutated to TAC that coded for tyrosine (F252Y). Conclusions: The present study identified for the first time a novel mutation within the conserved protein binding domain of IRF6 gene in tissue samples of subjects with OSCC.

• Animal Bioscience
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• v.35 no.6
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• pp.847-857
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• 2022

Objective: The effects of maternal undernutrition during midgestation on muscle fiber histology, myosin heavy chain (MyHC) expression, methylation modification of myogenic factors, and the mammalian target of rapamycin (mTOR) signaling pathway in the skeletal muscles of prenatal and postnatal goats were examined. Methods: Twenty-four pregnant goats were assigned to a control (100% of the nutrients requirement, n = 12) or a restricted group (60% of the nutrients requirement, n = 12) between 45 and 100 days of gestation. Descendants were harvested at day 100 of gestation and at day 90 after birth to collect the femoris muscle tissue. Results: Maternal undernutrition increased (p<0.05) the fiber area of the vastus muscle in the fetuses and enhanced (p<0.01) the proportions of MyHCI and MyHCIIA fibers in offspring, while the proportion of MyHCIIX fibers was decreased (p<0.01). DNA methylation at the +530 cytosine-guanine dinucleotide (CpG) site of the myogenic factor 5 (MYF5) promoter in restricted fetuses was increased (p<0.05), but the methylation of the MYF5 gene at the +274,280 CpG site and of the myogenic differentiation (MYOD) gene at the +252 CpG site in restricted kids was reduced (p<0.05). mTOR protein signals were down-regulated (p<0.05) in the restricted offspring. Conclusion: Maternal undernutrition altered the muscle fiber type in offspring, but its relationship with methylation in the promoter regions of myogenic genes needs to be elucidated.

• Fisheries and Aquatic Sciences
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• v.17 no.4
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• pp.471-478
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• 2014

Genomic organization, including the structural characteristics of 5'-flanking regions of two 65-kDa protein (WAP65) isoform genes associated with warm temperature acclimation, were characterized and their transcriptional responses to immune challenges were examined in the intestine, kidney and spleen of the mud loach (Misgurnus mizolepis; Cypriniformes). Both mud loach Wap65 isoform genes displayed a 10-exon structure that is common to most teleostean Wap65 genes. The two mud loach Wap65 isoforms were predicted to possess various stress- and immune-related transcription factor binding sites in their regulatory regions; however, the predicted motif profiles differed between the two isoforms, and the inflammation-related transcription factor binding motifs, such as NF-${\kappa}B$ and CREBP sites, were more highlighted in the Wap65-2 isoform than the Wap65-1 isoform. The results of qRT-PCR indicated that experimental immune challenges using Edwardsiella tarda, lipopolysaccharide or polyI:C induced the Wap65-2 isoform more than Wap65-1 isoform, although modulation patterns in response to these challenges were tissue- and stimulant-dependent. This study confirms that functional diversification between the two mud loach Wap65 isoforms (i.e., closer involvement of Wap65-2 in the acute phase of inflammation and innate immunity) occurs at the mRNA level in multiple tissues, and suggests that such differential modulation patterns between the two isoforms are related to the different transcription factor binding profiles in their regulatory regions.

• BMB Reports
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• v.44 no.3
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• pp.187-192
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• 2011

TGF-$\beta$ activated kinase-1 (TAK1) plays a pivotal role in developmental processes in many species. Previous research has mainly focused on the function of TAK1 in model organisms, and little is known about the function of TAK1 in hymenoptera insects. Here, we isolated and characterized the TAK1 gene from Apis cerana cerana. Promoter analysis of AccTAK1 revealed the presence of transcription factor binding sites related to early development. Real-time quantitative PCR and immunohistochemistry experiments revealed that AccTAK1 was expressed at high levels in fourth instar larvae, primarily in the abdomen, in the intestinal wall cells of the midgut and in the secretory cells of the salivary glands. In addition, AccTAK1 expression in fourth instar larvae could be dramatically induced by treatment with pesticides and organic solvents. These observations suggest that AccTAK1 may be involved in the regulation of early development in the larval salivary gland and midgut.

• Journal of Life Science
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• v.17 no.2 s.82
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• pp.167-173
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• 2007

To study the transcriptional mechanisms by which expression of the murine glial cell-derived neurotrophic factor inducible transcription factor (mGIF) gene is regulated, a murine genomic clone was iso-lated using a mGIF cDNA as probe. A 13-kb genomic fragment, which comprises 4-kb upstream of the transcription initiation site was sequenced. The promoter region lacks a TATA box and CAAT box, is rich in G+C content, and has multiple putative binding sites for the transcription factor Spl. The mGIF gene also has consensus sequences for AP2 binding sites. The transcriptional activity of five deletion mutants of a 2.1-kb fragment was analyzed by modulating transcription of the heterologous luciferase gene in the promoterless plasmid pGL2-Basic. All mutants showed significant transcriptional activity in the murine neuroblastoma cell line NB41A3. Transient expression assays suggested the presence of a positive regulator between -213 and -129 while a negative regulator was found in the region between -806 and -214. Relatively strong transcriptional activity was observed in neuronal NB41A3, glial C6 cells and hepatic HepG2, but very weak activity in skeletal muscle C2C12 cells. These findings confirm the tissue-specific activity of the mGIF promoter and suggest that this gene shares structural and functional similarities with the dopamine receptor genes that it regulates.