• Title/Summary/Keyword: Double regulating

Search Result 63, Processing Time 0.029 seconds

Accelerated Growth of Corynebacterium glutamicum by Up-Regulating Stress-Responsive Genes Based on Transcriptome Analysis of a Fast-Doubling Evolved Strain

  • Park, Jihoon;Lee, SuRin;Lee, Min Ju;Park, Kyunghoon;Lee, Seungki;Kim, Jihyun F.;Kim, Pil
    • Journal of Microbiology and Biotechnology
    • /
    • v.30 no.9
    • /
    • pp.1420-1429
    • /
    • 2020
  • Corynebacterium glutamicum, an important industrial strain, has a relatively slower reproduction rate. To acquire a growth-boosted C. glutamicum, a descendant strain was isolated from a continuous culture after 600 generations. The isolated descendant C. glutamicum, JH41 strain, was able to double 58% faster (td=1.15 h) than the parental type strain (PT, td=1.82 h). To understand the factors boosting reproduction, the transcriptomes of JH41 and PT strains were compared. The mRNAs involved in respiration and TCA cycle were upregulated. The intracellular ATP of the JH41 strain was 50% greater than the PT strain. The upregulation of NCgl1610 operon (a putative dyp-type heme peroxidase, a putative copper chaperone, and a putative copper importer) that presumed to role in the assembly and redox control of cytochrome c oxidase was found in the JH41 transcriptome. Plasmid-driven expression of the operon enabled the PT strain to double 19% faster (td=1.82 h) than its control (td=2.17 h) with 14% greater activity of cytochrome c oxidase and 27% greater intracellular ATP under the oxidative stress conditions. Upregulations of genes those might enhance translation fitness were also found in the JH41 transcriptome. Plasmid-driven expressions of NCgl0171 (encoding a cold-shock protein) and NCgl2435 (encoding a putative peptidyl-tRNA hydrolase) enabled the PT to double 22% and 32% faster than its control, respectively (empty vector: td=1.93 h, CspA: td=1.58 h, and Pth: td=1.44 h). Based on the results, the factors boosting growth rate in C. gluctamicum were further discussed in the viewpoints of cellular energy state, oxidative stress management, and translation.

The vacuolar processing enzyme (VPE) mutation suppresses an HR-like cell death induced by the double knockout mutant of vacuolar Ca2+-ATPases in Arabidopsis (애기장대에서 두 액포막 칼슘펌프 돌연변이에 의하여 유도되는 세포사멸 표현형의 액포수식효소(VPE) 돌연변이에 의한 억제)

  • Park, Hyeong-Cheol;Lee, Sang-Min;Kim, Ho-Soo;Chung, Woo-Sik
    • Journal of Plant Biotechnology
    • /
    • v.38 no.2
    • /
    • pp.169-175
    • /
    • 2011
  • Calcium ($Ca^{2+}$) signals have been implicated in regulating plant development and responses to the environmental stresses including a programmed cell death pathway. In animals and plants, cytosolic $Ca^{2+}$ signals have been involved in the activation of programmed cell death (PCD). Recently, we reported that disruption of Arabidopsis vacuolar $\b{A}$utoinhibited $\underline{C}a^{2+}$-$\b{A}$TPases (ACAs), ACA4 and ACA11, resulted in the activation of a salicylic acid-dependent programmed cell death pathway. Although extensive studies have revealed various components of a PCD in plants, executors to directly induce PCD are well unknown. Here, we provide that the vacuolar processing enzymes (VPEs) are involved in a PCD induced by the double knockout mutant of vacuolar $Ca^{2+}$-ATPases in Arabidopsis. The gene expression of VPE was rapidly up-regulated and the enzyme activity of VPE was increased in the double mutant plants. We also generated aca4/aca11/avpe, aca4/aca11/${\gamma}$vpe and aca4/aca11/avpe/${\gamma}$vpe mutant plants. Although cell death phenotype of the double mutant plants was not completely disappeared in the triple and quadruple mutant plants, the triple and quadruple mutant plants showed to significantly delay cell death phenotype of the double mutant plants. These results suggest that the VPE is involved in the HR-like cell death in the double mutant of vacuolar $Ca^{2+}$-ATPases in Arabidopsis.

Analysis of microRNA expression profiles during the cell cycle in synchronized HeLa cells

  • Zhou, Jue-Yu;Ma, Wen-Li;Liang, Shuang;Zeng, Ye;Shi, Rong;Yu, Hai-Lang;Xiao, Wei-Wei;Zheng, Wen-Ling
    • BMB Reports
    • /
    • v.42 no.9
    • /
    • pp.593-598
    • /
    • 2009
  • Cell cycle progression is regulated by both transcriptional and post-transcriptional mechanisms. MicroRNAs (miRNAs) emerge as a new class of small non-coding RNA regulators of cell cycle as recent evidence suggests. It is hypothesized that expression of specific miRNAs oscillates orderly along with cell cycle progression. However, the oscillated expression patterns of many candidate miRNAs have yet to be determined. Here, we describe miRNA expression profiling in double-thymidine synchronized HeLa cells as cell cycle progresses. Twenty-five differentially expressed miRNAs were classified into five groups based on their cell cycle-dependent expression patterns. The cyclic expression of six miRNAs (miR-221, let-7a, miR-21, miR-34a, miR-24, miR-376b) was validated by real-time quantitative RT-PCR (qRT-PCR). These results suggest that specific miRNAs, along with other key factors are required for maintaining and regulating proper cell cycle progression. The study deepens our understanding on cell cycle regulation.

Targeting HSP90 Gene Expression with 17-DMAG Nanoparticles in Breast Cancer Cells

  • Mellatyar, Hassan;Talaei, Sona;Nejati-Koshki, Kazem;Akbarzadeh, Abolfazl
    • Asian Pacific Journal of Cancer Prevention
    • /
    • v.17 no.5
    • /
    • pp.2453-2457
    • /
    • 2016
  • Background: Dysregulation of HSP90 gene expression is known to take place in breast cancer. Here we used D,L-lactic-co-glycolic acid-polyethylene glycol-17-dimethylaminoethylamino-17-demethoxy geldanamycin (PLGA-PEG-17DMAG) complexes and free 17-DMAG to inhibit the expression of HSP90 gene in the T47D breast cancer cell line. The purpose was to determine whether nanoencapsulating 17DMAG improves the anti-cancer effects as compared to free 17DMAG. Materials and Methods: The T47D breast cancer cell line was grown in RPMI 1640 supplemented with 10% FBS. Encapsulation of 17DMAG was conducted through a double emulsion method and properties of copolymers were characterized by Fourier transform infrared spectroscopy and H nuclear magnetic resonance spectroscopy. Assessment of drug cytotoxicity was by MTT assay. After treatment of T47D cells with a given amount of drug, RNA was extracted and cDNA was synthesized. In order to assess HSP90 gene expression, real-time PCR was performed. Results: Taking into account drug load, IC50 was significant decreased in nanocapsulated 17DMAG in comparison with free 17DMAG. This finding was associated with decrease of HSP90 gene expression. Conclusions: PLGA-PEG-17DMAG complexes can be more effective than free 17DMAG in down-regulating of HSP90 expression, at the saesm time exerting more potent cytotoxic effects. Therefore, PLGA-PEG could be a superior carrier for this type of hydrophobic agent.

Functional Roles of a Putative B' Delta Regulatory Subunit and a Catalytic Subunit of Protein Phosphatase 2A in the Cereal Pathogen Fusarium graminearum

  • Kim, Hee-Kyoung;Yun, Sung-Hwan
    • The Plant Pathology Journal
    • /
    • v.28 no.3
    • /
    • pp.259-269
    • /
    • 2012
  • Protein phosphatase 2A (PP2A), a family of serine/threonine protein phosphatases, plays an important role in balancing the phosphorylation status of cellular proteins for regulating diverse biological functions in eukaryotic organisms. Despite intensive studies in mammals, limited information on its role is available in filamentous fungi. Here, we investigated the functional roles of genes for a putative B' delta regulatory subunit (FgPP2AR) and a catalytic subunit (FgPP2AC) of PP2A in a filamentous ascomycete, Fusarium graminearum. Molecular characterization of an insertional mutant of this plant pathogenic fungus allowed us to identify the roles of FgPP2AR. Targeted gene replacement and complementation analyses demonstrated that the deletion of FgPP2AR, which was constitutively expressed in all growth stages, caused drastic changes in hyphal growth, conidia morphology/germination, gene expression for mycotoxin production, sexual development and pathogenicity. In particular, overproduction of aberrant cylindrical-shaped conidia is suggestive of arthroconidial induction in the ${\Delta}FgPP2AR$ strain, which has never been described in F. graminearum. In contrast, the ${\Delta}FgPP2AC$ strain was not significantly different from its wild-type progenitor in conidiation, trichothecene gene expression, and pathogenicity; however, it showed reduced hyphal growth and no perithecial formation. The double-deletion ${\Delta}FgPP2AR;{\Delta}FgPP2AC$ strain had more severe defects than single-deletion strains in all examined phenotypes. Taken together, our results indicate that both the putative regulatory and catalytic subunits of PP2A are involved in various cellular processes for fungal development in F. graminearum.

Molecular Cloning and Overexpression of Phytoene Desaturase (CrtI) from Paracoccus haeundaensis

  • Choi, Seong-Seok;Seo, Yong Bae;Lim, Han Kyu;Nam, Soo-Wan;Kim, Gun-Do
    • Microbiology and Biotechnology Letters
    • /
    • v.46 no.2
    • /
    • pp.145-153
    • /
    • 2018
  • Among the carotenoid biosynthesis genes, crtI gene encodes the phytoene desaturase (CrtI) enzyme, and phytoene desaturase convert phytoene to lycopene. Phytoene desaturase is involved in the dehydrogenation reaction, in which four single bonds in the phytoene are introduced into a double bond, eliminating eight hydrogen atoms in the process. Phytoene desaturase is one of the key regulating enzyme in carotenoid biosynthetic pathway of various carotenoid biosynthetic organisms. The crtI gene in genomic DNA of Paracoccus haeundaensis was amplified and cloned into a T-vector to analyze the nucleotide sequence. As a result, the crtI gene coding for phytoene desaturase from P. haeundaensis consists of 1,503 base pairs encoding 501 amino acids residues. An expression plasmid containing the crtI gene was constructed, and Escherichia coli cells containing this plasmid produced the recombinant protein of approximately 55 kDa, equivalent to the molecular weight of phytoene desaturase. The expressed protein in cell lysate showed enzymatic activity similar to phytoene desaturase. Phytoene and lycopene were analyzed by HPLC and measured at wavelength of 280 nm and 470 nm, respectively. The $K_m$ values for phytoene and NADPH were $11.1{\mu}M$ and $129.3{\mu}M$, respectively.

The Liability for Unsafe Medical Product and The Preemption Clause of Medical Device Act (의료기기의 결함으로 인한 손해배상책임과 미국 연방법 우선 적용 이론에 관하여)

  • Kim, Jang Han
    • The Korean Society of Law and Medicine
    • /
    • v.15 no.2
    • /
    • pp.63-89
    • /
    • 2014
  • In 1976, the Dalkon Shield-intrauterine device injured several thousand women in U.S.A. which caused the changes of medical deivce regulation. The Medical Device Regulation Act or Medical Device Amendments of 1976 (MDA) was introduce. As part of the process of regulating medical devices, the MDA divides medical devices into three categories. The class II, and III devices which have moderate harm or more can use the section 510 (k), premarket notification process if the manufacturer can establish that its device is "substantially equivalent" to a device that was marketed before 1976. In 21 U.S.C. ${\S}$ 360k(a), MDA introduced a provision which expressly preempts competing state laws or regulations. After that, the judicial debates had began over the proper interpretation and application of Section 360(k) In February 2008, the U.S. Supreme Court ruled in Riegel v. Medtronic that manufacturer approved by the Food and Drug Administration (FDA)'s pre-market approval process are preempted from liability, even when the devices have defective design or lack of labeling. But the Supreme Court ruled in Medtronic Inc. v. Lora Lohr that the manufactures which use the section 510 (k) process cannot be preempted and in Bausch v. Stryker Corp. that manufactures which violated the CGMP standard are also liable to the damage of patient at the state courts. In 2009, the Supreme Court ruled in Wyeth v. Levine that patients harmed by prescription drugs can claim damages in state courts. This may cause a double standard between prescription drugs and medical devices. FDA Preemption is the legal theory in the United States that exempts product manufacturers from tort claims regarding Food and Drug Administration approved products. FDA Preemption has been a highly contentious issue. In general, consumer groups are against it while the FDA and pharmaceutical manufacturers are in favor of it. This issues also influences the theory of product liability of U.S.A. Complete immunity preemption is an issue need to be more declared.

  • PDF

Expression of Calponin 3 in the Striatum Following 3-Nitropropionic Acid-induced Neurotoxicity (선조체에서 3-nitropropionic acid 투여 후 calponin 3의 발현 연구)

  • Choi, Yun-Sik
    • Journal of Life Science
    • /
    • v.23 no.1
    • /
    • pp.125-130
    • /
    • 2013
  • Calponin 3 is an F-actin-binding protein and plays a key role in regulating spine plasticity and synaptic activity in neurons. Unlike the other subtypes, calponin 1 and 2, which are expressed in smooth and cardiac muscle cells, calponin 3 is highly expressed in the brain. The goal of this study was to elucidate the spatiotemporal expression pattern of calponin 3 following repeated administration of 3-nitropropionic acid in mice. The repeated administration of 3-nitropropionic acid generated necrotic neuronal cell death in the striatum. Calponin 3 was up-regulated in the neuroprotective penimbral region from 1.5 days after the last injection and thereafter. Double immunofluorescence study revealed that calponin 3 was induced in GFAP-positive astrocytes. These results suggest that calponin 3 induction in the neuroprotective penumbral area following 3-nitropropionic acid intoxication may play a key role in reactive astrogliosis in the striatum.

A Study on Neuroactive Response Sensing Platform after Injection of Muscular Relaxants (근 이완제 투여에 따른 신경 자극 반응 감지 플랫폼에 관한 연구)

  • Kim, Woo-Ram;Kim, Young-Kil
    • Proceedings of the Korean Institute of Information and Commucation Sciences Conference
    • /
    • 2011.10a
    • /
    • pp.331-334
    • /
    • 2011
  • This is a study about a platform realization measuring the extent of reaction in nerve, as giving a electrical impulse on a nerve pulp regulating a function of muscle, about a measurement of nerve reaction in the amount of current, the lasting time of current, and the position of electrode from a electrical impuls.The position of an electrode in a electrical nerve impuls have nothing to do with all nerves from exercise to all things. There is the Single Twitch Stimulation(STS), Train-of-four(TOF), and Double Burst Stimulation(DBS) in the form of nerve stimulation. This report is needed for selecting MCU of low electric power for a base in embedded system and measuring the extent of reaction after making a sensor interface to know sensitivity of measuring sensor in basic reaction of nerve impuls. The platform is realized to select a high efficiency AD Convertor for raising accuracy in measured data. As the platform in this report was developed for a medical appliances, it was designed to consider user safety in electric power Isolation when making electric power circuit.

  • PDF

MAPK3 at the Autism-Linked Human 16p11.2 Locus Influences Precise Synaptic Target Selection at Drosophila Larval Neuromuscular Junctions

  • Park, Sang Mee;Park, Hae Ryoun;Lee, Ji Hye
    • Molecules and Cells
    • /
    • v.40 no.2
    • /
    • pp.151-161
    • /
    • 2017
  • Proper synaptic function in neural circuits requires precise pairings between correct pre- and post-synaptic partners. Errors in this process may underlie development of neuropsychiatric disorders, such as autism spectrum disorder (ASD). Development of ASD can be influenced by genetic factors, including copy number variations (CNVs). In this study, we focused on a CNV occurring at the 16p11.2 locus in the human genome and investigated potential defects in synaptic connectivity caused by reduced activities of genes located in this region at Drosophila larval neuromuscular junctions, a well-established model synapse with stereotypic synaptic structures. A mutation of rolled, a Drosophila homolog of human mitogen-activated protein kinase 3 (MAPK3) at the 16p11.2 locus, caused ectopic innervation of axonal branches and their abnormal defasciculation. The specificity of these phenotypes was confirmed by expression of wild-type rolled in the mutant background. Albeit to a lesser extent, we also observed ectopic innervation patterns in mutants defective in Cdk2, Gq, and Gp93, all of which were expected to interact with Rolled MAPK3. A further genetic analysis in double heterozygous combinations revealed a synergistic interaction between rolled and Gp93. In addition, results from RT-qPCR analyses indicated consistently reduced rolled mRNA levels in Cdk2, Gq, and Gp93 mutants. Taken together, these data suggest a central role of MAPK3 in regulating the precise targeting of presynaptic axons to proper postsynaptic targets, a critical step that may be altered significantly in ASD.