• Title/Summary/Keyword: Gene repression

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Ventx1.1 as a Direct Repressor of Early Neural Gene zic3 in Xenopus laevis

  • Umair, Zobia;Kumar, Shiv;Kim, Daniel H.;Rafiq, Khezina;Kumar, Vijay;Kim, SungChan;Park, Jae-Bong;Lee, Jae-Yong;Lee, Unjoo;Kim, Jaebong
    • Molecules and Cells
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    • v.41 no.12
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    • pp.1061-1071
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    • 2018
  • From Xenopus embryo studies, the BMP4/Smad1-targeted gene circuit is a key signaling pathway for specifying the cell fate between the ectoderm and neuro-ectoderm as well as the ventral and dorsal mesoderm. In this context, several BMP4/Smad1 target transcriptional factors have been identified as repressors of the neuro-ectoderm. However, none of these direct target transcription factors in this pathway, including GATA1b, Msx1 and Ventx1.1 have yet been proven as direct repressors of early neuro-ectodermal gene expression. In order to demonstrate that Ventx1.1 is a direct repressor of neuro-ectoderm genes, a genome-wide Xenopus ChIP-Seq of Ventx1.1 was performed. In this study, we demonstrated that Ventx1.1 bound to the Ventx1.1 response cis-acting element 1 and 2 (VRE1 and VRE2) on the promoter for zic3, which is a key early neuro-ectoderm gene, and this Ventx1.1 binding led to repression of zic3 transcription. Site-directed mutagenesis of VRE1 and VRE2 within zic3 promoter completely abolished the repression caused by Ventx1.1. In addition, we found both the positive and negative regulation of zic3 promoter activity by FoxD5b and Xcad2, respectively, and that these occur through the VREs and via modulation of Ventx1.1 levels. Taken together, the results demonstrate that the BMP4/Smad1 target gene, Ventx1.1, is a direct repressor of neuro-ectodermal gene zic3 during early Xenopus embryogenesis.

Transcriptional Repression of High-Mobility Group Box 2 by p21 in Radiation-Induced Senescence

  • Kim, Hyun-Kyung;Kang, Mi Ae;Kim, Mi-Sook;Shin, Young-Joo;Chi, Sung-Gil;Jeong, Jae-Hoon
    • Molecules and Cells
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    • v.41 no.4
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    • pp.362-372
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    • 2018
  • High mobility group box 2 (HMGB2) is an abundant, chromatin-associated, non-histone protein involved in transcription, chromatin remodeling, and recombination. Recently, the HMGB2 gene was found to be significantly downregulated during senescence and shown to regulate the expression of senescent-associated secretory proteins. Here, we demonstrate that HMGB2 transcription is repressed by p21 during radiation-induced senescence through the ATM-p53-p21 DNA damage signaling cascade. The loss of p21 abolished the downregulation of HMGB2 caused by ionizing radiation, and the conditional induction of p21 was sufficient to repress the transcription of HMGB2. We also showed that the p21 protein binds to the HMGB2 promoter region, leading to sequestration of RNA polymerase and transcription factors E2F1, Sp1, and p300. In contrast, NF-Y, a CCAAT box-binding protein complex, is required for the expression of HMGB2, but NF-Y binding to the HMGB2 promoter was unaffected by either radiation or p21 induction. A proximity ligation assay results confirmed that the chromosome binding of E2F1 and Sp1 was inhibited by p21 induction. As HMGB2 have been shown to regulate premature senescence by IR, targeting the p21-mediated repression of HMGB2 could be a strategy to overcome the detrimental effects of radiation-induced senescence.

OPTHiS Identifies the Molecular Basis of the Direct Interaction between CSL and SMRT Corepressor

  • Kim, Gwang Sik;Park, Hee-Sae;Lee, Young Chul
    • Molecules and Cells
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    • v.41 no.9
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    • pp.842-852
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    • 2018
  • Notch signaling is an evolutionarily conserved pathway and involves in the regulation of various cellular and developmental processes. Ligand binding releases the intracellular domain of Notch receptor (NICD), which interacts with DNA-bound CSL [CBF1/Su(H)/Lag-1] to activate transcription of target genes. In the absence of NICD binding, CSL down-regulates target gene expression through the recruitment of various corepressor proteins including SMRT/NCoR (silencing mediator of retinoid and thyroid receptors/nuclear receptor corepressor), SHARP (SMRT/HDAC1-associated repressor protein), and KyoT2. Structural and functional studies revealed the molecular basis of these interactions, in which NICD coactivator and corepressor proteins competitively bind to ${\beta}-trefoil$ domain (BTD) of CSL using a conserved ${\varphi}W{\varphi}P$ motif (${\varphi}$ denotes any hydrophobic residues). To date, there are conflicting ideas regarding the molecular mechanism of SMRT-mediated repression of CSL as to whether CSL-SMRT interaction is direct or indirect (via the bridge factor SHARP). To solve this issue, we mapped the CSL-binding region of SMRT and employed a 'one- plus two-hybrid system' to obtain CSL interaction-defective mutants for this region. We identified the CSL-interaction module of SMRT (CIMS; amino acid 1816-1846) as the molecular determinant of its direct interaction with CSL. Notably, CIMS contains a canonical ${\varphi}W{\varphi}P$ sequence (APIWRP, amino acids 1832-1837) and directly interacts with CSL-BTD in a mode similar to other BTD-binding corepressors. Finally, we showed that CSL-interaction motif, rather than SHARP-interaction motif, of SMRT is involved in transcriptional repression of NICD in a cell-based assay. These results strongly suggest that SMRT participates in CSL-mediated repression via direct binding to CSL.

Changes in Apoptosis-related Gene Expression Induced by Repression of FGFR1 by RNA Interference in Embryonic Fibroblasts and Cancerous Cells from Chicken

  • Lee, Sang-In;Lee, Bo-Ram;Hwang, Young-Sun;Rengaraj, Deivendran;Han, Jae-Yong
    • Journal of Animal Science and Technology
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    • v.52 no.6
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    • pp.521-527
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    • 2010
  • Fibroblast growth factor receptor 1 (FGFR1) plays roles in angiogenesis, wound healing, and embryonic development via the regulation of cell proliferation, differentiation, and survival. It is well known that ectopic expression of FGFR1 is associated with cancer development. To characterize the function of FGFR1 in the normal and cancer cell lines DF-1 and DT40, respectively, we performed FGFR1 knockdown by RNA interference. In the DT40 cells, FGFR1 knockdown induced upregulation of FGFR2 and FGFR3 expression, downregulation of pro-apoptosis-related genes, and upregulation of anti-apoptosis-related genes. However, in DF-1 cells, FGFR1 knockdown induced upregulation of pro-apoptosis-related genes and downregulation of anti-apoptosis-related genes. Our data suggest that repression of FGFR1 induced upregulation of other FGF receptors and anti-apoptosis-related genes in cancer cells and pro-apoptosis-related genes in normal cells.

Expressive Methods of Uncanny Image in Contemporary Fashion (현대패션에 나타난 언캐니 이미지의 표현방법)

  • Yu, Arim;Suh, Seunghee
    • Journal of the Korean Society of Costume
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    • v.65 no.6
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    • pp.99-111
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    • 2015
  • The purpose of this study is to identify the expression methods of uncanny images shown in contemporary fashion through the visual expression methods of plastic arts. The study results are as following. First, there is a concept-forming element of 'The return of Repressed', which is a psychological mechanism for generating uncanny sets to the component of 'repression' and 'return'. 'Repression' herein consisted of 'unconsciousness', 'the other', and 'anxiety', while the return was composed of 'trauma' and 'repetition compulsion'. Second, there are visual expression methods of the uncanny in contemporary art. The subject of 'repressed things' was expressed as grotesque and having a terrible look. 'Fragmented body' was analyzed as 'dismantling' by mutilation, 'combination' placing cut body parts in other places, and 'damage' suffering from an illness or accident. 'Fear of death' was shown as 'anatomy' reminiscent of death and 'iconography of death' meaning return of the dead. 'Post-human' was a surrealistic being such as a monster and mechanized human, and was analyzed as 'gene variation' and 'cyborg'. Third, the methods of visual expression of the uncanny reflected on contemporary fashion was analyzed as 'fragmented body', 'fear of death', and 'post-human'. 'Fragmented body' appeared as the object reminiscent of the fragmented body, 'dismantling and combination of the body', and 'damage of the body' through the distortion of the clothing construction principle. 'Fear of death' visualized the characteristic elements of the subject reminiscent of death as 'iconography of death' and 'symbolic object of death' through the motif. 'Post-human' was the description of the subject beyond the body's function and form, and was shown as 'gene variation' and 'cyborg'.

Effect of Temperature and Carbon Source on the Expression of $\beta$-Galactosidase Gene of Lactococcus lactis ssp. lactis ATCC 7962

  • Kim, Tea-Youn;Lee, Jung-Min;Chang, Hae-Choon;Chung, Dae-Kyun;Lee, Jong-Hoon;Kim, Jeong-Hwan;Lee, Hyong-Joo
    • Journal of Microbiology and Biotechnology
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    • v.9 no.2
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    • pp.201-205
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    • 1999
  • The effects of growth temperature and a carbon source on the expression of $\beta$-galactosidase gene of Lactococcus lactis ssp. lactis ATCC 7962 (L. lactis 7962) were investigated. At $25^{\circ}C$, L. lactis 7962 had a higher $\beta$-galactosidase activity than cells grown at $30^{\circ}C$ or $37^{\circ}C$, although cells grew most quickly at $37^{\circ}C$ The highest $\beta$-galactosidase activity was observed in cells grown in M17 with lactose (l %) followed by cells grown in a galactose (1 %) medium. L. lactis 7962 exhibited the minimum $\beta$-galactosidase activity in glucose media, indicating catabolite repression. When the cellular levels of $\beta$-galactosidase mRNA were examined using slot blot hybridization, no significant differences were observed between cells grown at $25^{\circ}C$ and cells at $30^{\circ}C$ or $37^{\circ}C$ in the same media. This suggests that the quantity of $\beta$-galactosidase mRNA may not be the reason for the higher $\beta$-galactosidase activities of L. lactis 7962 at $25^{\circ}C$ The level of ccpA (Catabolite Control Protein) transcript remained almost constant during the exponential growth phase irrespective of a carbon sourse.

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Effects of the Repression of sphpr1 Expression on Growth and mRNA Export in Fission Yeast (분열효모에서 sphpr1 유전자의 결실이 생장 및 mRNA Export에 미치는 영향)

  • Lee, Hyun-Joo;Yoon, Jin-Ho
    • Korean Journal of Microbiology
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    • v.48 no.2
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    • pp.171-174
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    • 2012
  • THOC1/Hpr1 is one subunit of THO complex that is an evolutionally conserved assembly involved in the mRNP packaging and mRNA export during transcription elongation. In fission yeast Schizosaccharomyces pombe, an ortholog (spHpr1) of THOC1/Hpr1 was identified based on sequence alignment. A deletion mutant in a diploid strain was constructed by replacing one of spHpr1-coding region with a $kan^r$ gene using one-step gene disruption method. Tetrad analysis showed that the sphpr1 is essential for growth. Over-expression of sphpr1 from strong nmt1 promoter caused no defects of growth and mRNA export. However, repression of the sphpr1 expression resulted in growth inhibition accompanied by accumulation of poly$(A)^+$ RNA in the nucleus. These results suggest that spHpr1 is involved in mRNA export from the nucleus to cytoplasm.

A Yeast MRE3/REC114 Gene is Essential for Normal Cell Growth and Meiotic Recombination

  • Leem, Sun-Hee
    • Journal of Microbiology
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    • v.37 no.4
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    • pp.248-255
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    • 1999
  • We have analyzed the MRE3/REC114 gene of Saccharomyces cerevisiae, previously detected in isolation of mutants defective in meiotic recombination. We cloned the MRE3/REC114 gene by complementation of the meiotic recombination defect and it has been mapped to chormosome XIII. The DNA sequence analysis revealed that the MRE3 gene is identical to the REC114 gene. The upstream region of the MRE3/REC114 gene contains a T_4C site, a URS (upstream repression sequence) and a TR (T-rich) box-like sequence, which reside upstream of many meiotic genes. Coincidentally, northern blot analysis indicated that the three sizes of MRE3/REC114 transcripts, 3.4, 1.4 and 1.2 kb, are induced in meiosis. A less abundant transcript of 1.4 kb is detected in both mitotic and meiotic cells, suggesting that it is needed in mitosis as well as meiosis. To examine the role of the MRE3/REC114 gene, we constructed mre3 disruption mutants. Strains carrying an insertion or null deletion of the MRE3/REC114 gene showed slow growth in nutrient medium and the doubling time of these cells increased approximately by 2-fond compared to the wild-type strain. Moreover, the deletion mutant (${\delta}$mre3) displayed no meiotically induced recombination and no viable spores. The mre3/rec114 spore lethality can be suppressed by spo13, a mutation that causes cells to bypass reductional division. The double-stranded breaks (DSBs) which are involved in initiation of meiotic recombination were not detected in the analysis of meiotic chromosomal DNA from the mre3/rec114 disruptant. From these results we suggest that the MRE3/REC114 gene product is essential in normal growth and in early meiotic stages involved in meiotic recombination.

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REPRESSION OF Lhcb GENES FOR CHLOROPHYLL a/b-BINDING PROTEINS UNDER HIGH-LIGHT CONDITIONS IN Chlamydomonas

  • Haruhiko Teramoto;Akira Nakamori;Jun Minagawa;Ono, Taka-aki
    • Journal of Photoscience
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    • v.9 no.2
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    • pp.373-375
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    • 2002
  • Lhcb genes encoding light-harvesting chlorophyll-a/b binding (LHC) proteins of photosystem (PS) II were comprehensively characterized using the expressed sequence tag (EST) databases in the green alga, Chlamydomonas reinhardtii. The gene family was composed of eight Lhcb genes including four new genes, which were isolated and sequenced. The effects of light intensity on the levels of mRNAs accumulation of multiple Lhcb genes were studied under various conditions. The results indicate that Lhcb genes are coordinately regulated in response to light conditions, and repressed when the input light energy exceeded the requirement for $CO_2$ assimilation. The effects of high light on the expression of the Lhcb genes observed in the presence of an electron transport inhibitor, DCMU, and in mutants deficient in photosynthetic reaction centers suggest the presence of two alternative mechanisms for regulating the genes expression under high-light conditions.

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A Gene of Neurospora crassa that Encodes a Protein Containing TPR Motifs

  • Lee Bheong-Uk
    • Proceedings of the Microbiological Society of Korea Conference
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    • 2003.05a
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    • pp.51-54
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    • 2003
  • Analysis of the Neurospora crassa genome data reveals at least 14 proteins that contain tetratricopeptide repeat (TPR) motifs. One of them shows over $60\%$ homology with SSN6 of Saccharomyces cerevisiae, a global repressor that mediates repression of genes involved in various cellular processes. Sequence analysis of its cDNA shows that it encodes a putative 102kDa protein. Mutant strains generated by RIP (repeat induced point mutation) process show four distinctive patterns of vegetative growth at various rates. They are male-fertile, yet all female-sterile and produced little or no perithecium. These results indicate that this gene is pleiotropic and involved in several cellular processes of vegetative growth, conidiation and sexual cycle. It is designated rcm-1(regulation of conidiation and morphology).

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