• Title/Summary/Keyword: Molecular genetics

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Cyclic AMP response element binding (CREB) protein acts as a positive regulator of SOX3 gene expression in NT2/D1 cells

  • Kovacevic-Grujicic, Natasa;Mojsin, Marija;Popovic, Jelena;Petrovic, Isidora;Topalovic, Vladanka;Stevanovic, Milena
    • BMB Reports
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    • v.47 no.4
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    • pp.197-202
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    • 2014
  • SOX3 is one of the earliest neural markers in vertebrates, playing the role in specifying neuronal fate. In this study we have established first functional link between CREB and human SOX3 gene which both have important roles in the nervous system throughout development and in the adulthood. Here we demonstrate both in vitro and in vivo that CREB binds to CRE half-site located -195 to -191 within the human SOX3 promoter. Overexpression studies with CREB or its dominant-negative inhibitor A-CREB indicate that this transcription factor acts as a positive regulator of basal SOX3 gene expression in NT2/D1 cells. This is further confirmed by mutational analysis where mutation of CREB binding site results in reduction of SOX3 promoter activity. Our results point at CREB as a positive regulator of SOX3 gene transcription in NT2/D1 cells, while its contribution to RA induction of SOX3 promoter is not prominent.

Downregulation of Foxe1 by HR suppresses Msx1 expression in the hair follicles of HrHp mice

  • Choi, Jee-Hyun;Kim, Byong-Kyu;Kim, Jeong-Ki;Lee, Hwa-Young;Park, Jong-Keun;KimYoon, Sung-Joo
    • BMB Reports
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    • v.44 no.7
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    • pp.478-483
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    • 2011
  • Hairless (HR), a transcriptional cofactor, is highly expressed in the skin and brain. To characterize the effects of HR expression in the skin, we examined its capacity for transcriptional regulation of its target genes in mouse skin and keratinocytes. We found that Foxe1 mRNA expression was suppressed in HR-overexpressing skin, as well as in HR-expressing keratinocytes. In turn, Msx1 expression was downregulated contingent on Foxe1 downregulation in skin and keratinocytes. We also found that expression of Sfrp1 was also correlated with that of Foxe1. Further investigation of the mechanisms involved in the transcriptional regulation of these genes will facilitate our understanding of the relationship among genes involved in hair follicle morphogenesis and cycling.

PCR-Based Sensitive Detection of Wood-Decaying Fungus Phellinus linteus by Specific Primer from rDNA ITS Regions

  • Park, Dong-Suk;Kang, Hee-Wan;Kim, Ki-Tae;Cho, Soo-Muk;Park, Young-Jin;Shin, Hye-Sun;Lee, Byoung-Moo;Go, Seung-Joo
    • Mycobiology
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    • v.29 no.1
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    • pp.7-10
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    • 2001
  • Based on the rDNA ITS sequences data, specific primer set for PCR detection of wood-decaying fungus Phellinus linteus was designed. The length of PCR products using designed primer set(SHF and SHR) was about 540 bp. Among 11 species, 17 isolates of Phellinus spp. including Phellinus linteus, P. pomaceus, P. spiculosus, P. baumi, P. pini, P. igniarius, P. gilvus, P. biscuspidatus, P. weirii, P. johnsonianus, P. robutus, and P. igniarius, seven isolates of Phellinus linteus showed about 540 bp-sized single band. This molecular technique could offer a useful tool for detecting and identifying Phellinus linteus.

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mTOR signalling pathway - A root cause for idiopathic autism?

  • Ganesan, Harsha;Balasubramanian, Venkatesh;Iyer, Mahalaxmi;Venugopal, Anila;Subramaniam, Mohana Devi;Cho, Ssang-Goo;Vellingiri, Balachandar
    • BMB Reports
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    • v.52 no.7
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    • pp.424-433
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    • 2019
  • Autism spectrum disorder (ASD) is a complex neurodevelopmental monogenic disorder with a strong genetic influence. Idiopathic autism could be defined as a type of autism that does not have a specific causative agent. Among signalling cascades, mTOR signalling pathway plays a pivotal role not only in cell cycle, but also in protein synthesis and regulation of brain homeostasis in ASD patients. The present review highlights, underlying mechanism of mTOR and its role in altered signalling cascades as a triggering factor in the onset of idiopathic autism. Further, this review discusses how distorted mTOR signalling pathway stimulates truncated translation in neuronal cells and leads to downregulation of protein synthesis at dendritic spines of the brain. This review concludes by suggesting downstream regulators such as p70S6K, eIF4B, eIF4E of mTOR signalling pathway as promising therapeutic targets for idiopathic autistic individuals.

Expression of PACT and EIF2C2, Implicated in RNAi and MicroRNA Pathways, in Various Human Cell Lines

  • Lee, Yong-Sun;Jeon, Yesu;Park, Jong-Hoon;Hwang, Deog-Su;Dutta, Anindya
    • Animal cells and systems
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    • v.8 no.3
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    • pp.213-220
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    • 2004
  • MicroRNA and siRNA (small interfering RNA), representative members of small RNA, exert their effects on target gene expression through association with protein complexes called miRNP (microRNA associated ribonucleoproteins) and RISC (RNA induced silencing complex), respectively. Although the protein complexes are yet to be fully characterized, human EIF2C2 protein has been identified as a component of both miRNP and RISC. In this report, we raised antiserum against EIF2C2 in order to begin understanding the protein complexes. An immunoblot result indicates that EIF2C2 protein is ubiquitously expressed in a variety of cell lines from human and mouse. EIF2C2 protein exists in both cellular compartments, as indicated by an immunoblot assay with a nuclear extract and a cytosolic fraction (S100 fraction) from HeLa S3 lysate. Depletion of EIF2C1 or EIF2C2 protein resulted in a decrease of microRNA, suggesting a possible role of these proteins in microRNA stability or biogenesis. We also prepared antiserum against dsRNA binding protein PACT, whose homologs in C. elegans and Drosophila are known to have a role in the RNAi (RNA interference) pathway. The expression of PACT protein was also observed in a wide range of cell lines.

Expression Analysis of the Mx Gene and Its Genome Structure in Chickens

  • Yin, C.G.;Du, L.X.;Li, S.G.;Zhao, G.P.;Zhang, J.;Wei, C.H.;Xu, L.Y.;Liu, T.;Li, H.B.
    • Asian-Australasian Journal of Animal Sciences
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    • v.23 no.7
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    • pp.855-862
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    • 2010
  • Among the known interferon-induced antiviral mechanisms, the Mx pathway is one of the most powerful pathways. The Mx protein has direct antiviral activity and inhibits a wide range of viruses by blocking an early stage of the viral replication cycle. Cloning, characterization, and expression of Mx in vivo and in vitro have been conducted. The chicken Mx gene spans 21 kb and is made up of 14 exons and 13 introns, of which the promoter region was analyzed. The real-time PCR results showed that Mx expression was increased in chicken embryo fibroblasts (CEF) after 12- and 24-h induction with polyI: C. Induction of Mx expression by poly I: C in vivo revealed tissue-specific patterns among the chicken tissues tested. A trace expression of Mx was detected in healthy chicken liver tissues from adult chickens without inducement; the expression levels in the liver, heart, and gizzard were higher than in the muscle and kidney. This is the first report to demonstrate the expression of a glutathione-S-transferase-tagged-Mx fusion protein of 75 KDa, as well as the biological activity tested by SDS-PAGE and western blotting.