• Title/Summary/Keyword: transcriptional silencing

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Stearoyl-CoA desaturase induces lipogenic gene expression in prostate cancer cells and inhibits ceramide-induced cell death

  • Kim, Seung-Jin;Kim, Eung-Seok
    • Animal cells and systems
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    • v.15 no.1
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    • pp.1-8
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    • 2011
  • Perturbation of metabolism with increased expression of lipogenic enzymes is a common characteristic of human cancers, including prostate cancer. In the present work the overexpression of stearoyl-CoA desaturase (SCD) in LNCaP cells led to increased mRNA levels of fatty acid synthase (FAS) and acetyl-CoA-carboxylase-a, whereas micro RNA-mediated silencing of SCD inhibited the expression of these lipogenic genes in LNCaP cells. Treatment with the FAS-specific inhibitor cerulenin inhibited SCD induction of LNCaP cell proliferation. In addition, a transient transfection assay revealed the capability of cerulenin to suppress SCD and dihydrotestosterone induction of androgen receptor transcriptional activity. Furthermore, overexpression of SCD in LNCaP cells produced marked resistance to ceramide-induced cell death with reduced poly(ADP-ribose) polymerase (PARP) cleavage. In contrast, silencing of SCD expression increased Bax protein in LNCaP cells. Furthermore, addition of ceramide to SCD knockdown LNCaP cells increased cell death and caspase-3 activity with drastic increase of PARP cleavage. Together, the data indicate that SCD may provide resistance of prostate cancer cells to ceramide-induced cell death.

Gene Silencing Induced by Cytosine Methylation in Transgenic Tomato (형질전환 토마토에서 Cytosine Methylation에 의한 유전자발현 억제)

  • Jung, Seo-Hee;Min, Sung-Ran;Lee, Soo-Young;Park, Ji-Young;Davarpanah, S Javad;Chung, Hwa-Jee;Jeon, Jae-Heung;Liu, Jang-Ryol;Jeong, Won-Joong
    • Journal of Plant Biotechnology
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    • v.34 no.4
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    • pp.323-329
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    • 2007
  • Transgene expression was analyzed in tomato plants. Four lines of neomycin phosphotransferase II gene (NPTII) and the trehalose biosynthetic fusion gene (TPSP) transformed $T_0$ plants showed kanamycin resistance on selection medium. However, the analysis of phenotype (kanamycin resistance) and mRNA expression in $T_1$ plants indicated that the expression of the NPTII and TPSP transgenes was down-regulated to an undetectable level in two independent lines 1 and 11. Southern analysis demonstrated that the lines 1 and 11 had multicopies of the transgenes, whereas the typical transgenic lines 2 and 10 had 1 or 2 copies. DNA methylation analysis using methylation sensitive enzyme detected accumulated CpG DNA methylation on TPSP coding region and CaMV35S promoter region in the line 11, but not the typical transgenic line 2. These results suggest that multicopy transgene in plants is attributed to down-regulation of the transgene expression via transcriptional gene silencing.

Role of telomere length in subtelomeric gene expression and its possible relation to cellular senescence

  • Hernandez-Caballero, E.;Herrera-Gonzalez, N.E.;Salamanca-Gomez, F.;Arenas-Aranda, D.J.
    • BMB Reports
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    • v.42 no.11
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    • pp.747-751
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    • 2009
  • Transcriptional silencing of subtelomeric genes is associated with telomere length, which is correlated with age. Long and short telomeres in young and old people, respectively, coincide with gene repression and activation in each case. In addition, differential location of genes with respect to telomeres causes telomere position effect. There is very little evidence of the manner in which age-related telomere length affects the expression of specific human subtelomeric genes. We analyzed the relationship between telomere length and gene expression levels in fibroblasts derived from human donors at ages ranging from 0-70 years. We studied three groups of genes located between 100 and 150 kb, 200 and 250 kb, and >300 kb away from telomeres. We found that the chromatin modifier-encoding genes Eu-HMTase1, ZMYND11, and RASA3 were overexpressed in adults. Our results suggest that short telomere length-related overexpression of chromatin modifiers could underlie transcriptional changes contributing to cellular senescence.

Setdb1 Is Required for Myogenic Differentiation of C2C12 Myoblast Cells via Maintenance of MyoD Expression

  • Song, Young Joon;Choi, Jang Hyun;Lee, Hansol
    • Molecules and Cells
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    • v.38 no.4
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    • pp.362-372
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    • 2015
  • Setdb1, an H3-K9 specific histone methyltransferase, is associated with transcriptional silencing of euchromatic genes through chromatin modification. Functions of Setdb1 during development have been extensively studied in embryonic and mesenchymal stem cells as well as neurogenic progenitor cells. But the role of Sedtdb1 in myogenic differentiation remains unknown. In this study, we report that Setdb1 is required for myogenic potential of C2C12 myoblast cells through maintaining the expressions of MyoD and muscle-specific genes. We find that reduced Setdb1 expression in C2C12 myoblast cells severely delayed differentiation of C2C12 myoblast cells, whereas exogenous Setdb1 expression had little effect on. Gene expression profiling analysis using oligonucleotide microarray and RNA-Seq technologies demonstrated that depletion of Setdb1 results in downregulation of MyoD as well as the components of muscle fiber in proliferating C2C12 cells. In addition, exogenous expression of MyoD reversed transcriptional repression of MyoD promoter-driven luciferase reporter by Setdb1 shRNA and rescued myogenic differentiation of C2C12 myoblast cells depleted of endogenous Setdb1. Taken together, these results provide new insights into how levels of key myogenic regulators are maintained prior to induction of differentiation.

When a ribosome encounters a premature termination codon

  • Hwang, Jungwook;Kim, Yoon Ki
    • BMB Reports
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    • v.46 no.1
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    • pp.9-16
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    • 2013
  • In mammalian cells, aberrant transcripts harboring a premature termination codon (PTC) can be generated by abnormal or inefficient biogenesis of mRNAs or by somatic mutation. Truncated polypeptides synthesized from these aberrant transcripts could be toxic to normal cellular functions. However, mammalian cells have evolved sophisticated mechanisms for monitoring the quality of mRNAs. The faulty transcripts harboring PTC are subject to nonsense-mediated mRNA decay (NMD), nonsense-mediated translational repression (NMTR), nonsense-associated alternative splicing (NAS), or nonsense-mediated transcriptional gene silencing (NMTGS). In this review, we briefly outline the molecular characteristics of each pathway and suggest mRNA quality control mechanisms as a means to regulate normal gene expression.

Neuronal Activity-Dependent Regulation of MicroRNAs

  • Sim, Su-Eon;Bakes, Joseph;Kaang, Bong-Kiun
    • Molecules and Cells
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    • v.37 no.7
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    • pp.511-517
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    • 2014
  • MicroRNAs are non-coding short (~23 nucleotides) RNAs that mediate post-transcriptional regulation through sequence-specific gene silencing. The role of miRNAs in neuronal development, synapse formation and synaptic plasticity has been highlighted. However, the role of neuronal activity on miRNA regulation has been less focused. Neuronal activity-dependent regulation of miRNA may finetune gene expression in response to synaptic plasticity and memory formation. Here, we provide an overview of miRNA regulation by neuronal activity including high-throughput screening studies. We also discuss the possible molecular mechanisms of activity-dependent induction and turnover of miRNAs.

Epigenetic Regulation of Plant Reproductive Development

  • Vyskot, Boris
    • Korean Journal of Plant Tissue Culture
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    • v.27 no.5
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    • pp.359-366
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    • 2000
  • Epigenetics represents a chromatin-mediated transcriptional repression which plays a control role in both animal and plant development. A number of different mechanisms have been described to be involved in the formation of chromatin structure: especially DNA methylation, nucleosomal histone modification, DNA replication timing, and binding of chromatin remodelling proteins. Epigenetic phenomena include genomic imprinting, dosage compensation of X-chromosome linked genes, mutual allelic interactions, paramutation, transvection, silencing of invasive DNA sequences, etc. They are often unstable and inherited in a non-Mendelian way. A number of epigenetic defects has been preferentially described in floral development. Here, epigenetic phenomena in model angiosperm plants and their corresponding mechanisms are reviewed.

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Relationship Between Plant Viral Encoded Suppressor to Post-transcriptional Gene Silencing and Elicitor to R Gene-specific Host Resistance

  • Park, Chang-Won;Feng Qu;Tao Ren;T. Jack Morris
    • The Plant Pathology Journal
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    • v.20 no.1
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    • pp.22-29
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    • 2004
  • Many important horticultural and field crops are susceptible to virus infections or may possess a degree of resistance to some viruses, but become infected by others. Plant viruses enter cells through the presence of wounds, and replicate intracellularly small genomes that encode genes required for replication, cell-to-cell movement and encapsidation. There are numerous evidences from specific virus-host interactions to require the involvement of host factors and steps during viral replication cycle. However, viruses should deal with host defense responses either by general or specific mechanisms, targeting viral components or genome itself. On the other hand, the host plants have also adapted to defend themselves against viral attack by operating different lines of resistance responses. The defense-related interactions provide new insights into the complex molecular strategies for hosts for defense and counter-defense employed by viruses.

shRNA Mediated RHOXF1 Silencing Influences Expression of BCL2 but not CASP8 in MCF-7 and MDA-MB-231 Cell Lines

  • Ghafouri-Fard, Soudeh;Abdollahi, Davood Zare;Omrani, Mirdavood;Azizi, Faezeh
    • Asian Pacific Journal of Cancer Prevention
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    • v.13 no.11
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    • pp.5865-5869
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    • 2012
  • RHOXF1 has been shown to be expressed in embryonic stem cells, adult germline stem cells and some cancer lines. It has been proposed as a candidate gene to encode transcription factors regulating downstream genes in the human testis with antiapoptotic effects. Its expression in cancer cell lines has implied a similar role in the process of tumorigenesis. The human breast cancer cell lines MDA-MB-231 and MCF-7 were cultured in DMEM medium and transfected with a pGFP-V-RS plasmid bearing an RHOXF1 specific shRNA. Quantitative real-time RT-PCR was performed for RHOXF1, CASP8, BCL2 and HPRT genes. Decreased RHOXF1 expression was confirmed in cells after transfection. shRNA knock down of RHOXF1 resulted in significantly decreased BCL2 expression in both cell lines but no change in CASP8 expression. shRNA targeting RHOXF1 was shown to specifically mediate RHOXF1 gene silencing, so RHOXF1 can mediate transcriptional activation of the BCL2 in cancers and may render tumor cells resistant to apoptotic cell death induced by anticancer therapy. shRNA mediated knock down of RHOXF1 can be effective in induction of apoptotic pathway in cancer cells via BCL2 downregulation, so it can have potential therapeutic utility for human breast cancer.

Characterization of a Tomato (Lycopersicon esculentum Mill.) Ripening-associated Membrane Protein (TRAMP) Gene Expression and Flavour Volatile Changes in TRAMP Transgenic Plants

  • Kim Seog-Hyung;Ji Hee-Chung;Lim Ki-Byung
    • Journal of Plant Biotechnology
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    • v.7 no.2
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    • pp.87-95
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    • 2005
  • The tomato ripening associated membrane protein (TRAMP) (Fray et al., 1994) is a member of the major intrinsic protein (MIP) family, defined as channels facilitating the passage of water and small solutes through membranes. During normal fruit ripening the TRAMP mRNA levels were increased whereas the expression levels of TRAMP in low ethylene ACO1-sense suppressed lines, Nr and rin fruits, were lower than at the breaker stage of wild type fruit. TRAMP mRNA is inhibited by $LaCl_3$, which is an inhibitor of $Ca^{2+}$-stimulated responses, treatment but drought condition did not affect TRAMP expression. The levels of TRAMP mRNA transcripts were substantially higher in the dark treated seedlings and fruits. These suggest that TRAMP function as a water channel may be doubted because of several reasons; no water content was changed during ripening in wild type, antisense and overexpression lines, TRAMP expression under light condition was lower than dark condition and TRAMP expression was not changed in drought condition. Co-suppression plant, 3588 was one of sense suppression lines, which contain CaMV 35S promoter and sense pNY507 cDNA, produced small antisense RNA, approximately 21-25 nucleotides in length, mediated post-transcriptional gene silencing. Therefore, TRAMP expression was inhibited by small antisense and multiple copies might induce gene silencing without any production of double strand RNA. Total seven selected volatile productions, isobutylthiazole, 6-methyl-5-hepten-2-one, hexanal, hexenal methylbutanal, hexenol, and methylbutanol, were highly reduced in sense line whereas total volatile production was increased in TRAMP antisense line. These results suggested TRAMP might change volatile related compounds.