• Title/Summary/Keyword: molecular processes

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Post-Translational Regulation of the RSF1 Chromatin Remodeler under DNA Damage

  • Min, Sunwoo;Choi, Yong Won;Yun, Hansol;Jo, Sujin;Ji, Jae-Hoon;Cho, Hyeseong
    • Molecules and Cells
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    • v.41 no.2
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    • pp.127-133
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    • 2018
  • Chromatin remodeling factors are involved in many cellular processes such as transcription, replication, and DNA damage response by regulating chromatin structure. As one of chromatin remodeling factors, remodeling and spacing factor 1 (RSF1) is recruited at double strand break (DSB) sites and regulates ataxia telangiectasia mutated (ATM) -dependent checkpoint pathway upon DNA damage for the efficient repair. RSF1 is overexpressed in a variety of cancers, but regulation of RSF1 levels remains largely unknown. Here, we showed that protein levels of RSF1 chromatin remodeler are temporally upregulated in response to different DNA damage agents without changing the RSF1 mRNA level. In the absence of SNF2h, a binding partner of RSF1, the RSF1 protein level was significantly diminished. Intriguingly, the level of RSF1-3SA mutant lacking ATM-mediated phosphorylation sites significantly increased, and upregulation of RSF1 levels under DNA damage was not observed in cells overexpressing ATM kinase. Furthermore, failure in the regulation of RSF1 level caused a significant reduction in DNA repair, whereas reconstitution of RSF1, but not of RSF1-3SA mutants, restored DSB repair. Our findings reveal that temporal regulation of RSF1 levels at its post-translational modification by SNF2h and ATM is essential for efficient DNA repair.

Identification of the Antidepressant Vilazodone as an Inhibitor of Inositol Polyphosphate Multikinase by Structure-Based Drug Repositioning

  • Lee, Boah;Park, Seung Ju;Lee, Seulgi;Park, Seung Eun;Lee, Eunhye;Song, Ji-Joon;Byun, Youngjoo;Kim, Seyun
    • Molecules and Cells
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    • v.43 no.3
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    • pp.222-227
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    • 2020
  • Inositol polyphosphate multikinase (IPMK) is required for the biosynthesis of inositol phosphates (IPs) through the phosphorylation of multiple IP metabolites such as IP3 and IP4. The biological significance of IPMK's catalytic actions to regulate cellular signaling events such as growth and metabolism has been studied extensively. However, pharmacological reagents that inhibit IPMK have not yet been identified. We employed a structure-based virtual screening of publicly available U.S. Food and Drug Administration-approved drugs and chemicals that identified the antidepressant, vilazodone, as an IPMK inhibitor. Docking simulations and pharmacophore analyses showed that vilazodone has a higher affinity for the ATP-binding catalytic region of IPMK than ATP and we validated that vilazodone inhibits IPMK's IP kinase activities in vitro. The incubation of vilazodone with NIH3T3-L1 fibroblasts reduced cellular levels of IP5 and other highly phosphorylated IPs without influencing IP4 levels. We further found decreased Akt phosphorylation in vilazodone-treated HCT116 cancer cells. These data clearly indicate selective cellular actions of vilazodone against IPMK-dependent catalytic steps in IP metabolism and Akt activation. Collectively, our data demonstrate vilazodone as a method to inhibit cellular IPMK, providing a valuable pharmacological agent to study and target the biological and pathological processes governed by IPMK.

Rapamycin-Induced Abundance Changes in the Proteome of Budding Yeast

  • Shin, Chun-Shik;Chang, Yeon-Ji;Lee, Hun-Goo;Huh, Won-Ki
    • Genomics & Informatics
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    • v.7 no.4
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    • pp.203-207
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    • 2009
  • The target of rapamycin (TOR) signaling pathway conserved from yeast to human plays critical roles in regulation of eukaryotic cell growth. It has been shown that TOR pathway is involved in several cellular processes, including ribosome biogenesis, nutrient response, autophagy and aging. However, due to the functional diversity of TOR pathway, we do not know yet some key effectors of the pathway. To find unknown effectors of TOR signaling pathway, we took advantage of a green fluorescent protein (GFP)-tagged collection of budding yeast Saccharomyces cerevisiae. We analyzed protein abundance changes by measuring the GFP fluorescence intensity of 4156 GFP-tagged yeast strains under inhibition of TOR pathway. Our proteomic analysis argues that 83 proteins are decreased whereas 32 proteins are increased by treatment of rapamycin, a specific inhibitor of TOR complex 1 (TORC1). We found that, among the 115 proteins that show significant changes in protein abundance under rapamycin treatment, 37 proteins also show expression changes in the mRNA levels by more than 2-fold under the same condition. We suggest that the 115 proteins indentified in this study may be directly or indirectly involved in TOR signaling and can serve as candidates for further investigation of the effectors of TOR pathway.

Improvement of Functional Recovery by Cell Transplantation after Spinal Cord Injury (척수손상 후 세포이식에 의한 운동기능의 회복증진)

  • 이배환;이경희;성제경;황세진;김계성
    • Science of Emotion and Sensibility
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    • v.7 no.2
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    • pp.179-186
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    • 2004
  • Acute spinal cord injury can produce neurologic injury with many physical, psychological and social ramifications. It has been shown that two separate components combine to produce neurologic damage in acute spinal cord injury : the primary and secondary injuries. The primary mediators of spinal cord injury include the actual mechanical tissue disruption which is a passive process that occurs immediately following the trauma. A secondary injury cascade follows which appears mediated by cellular and molecular processes working through complex mechanisms. Both the primary and secondary injury cascades produce cell death both in neuronal and supporting cell tissues. Recovery from central nervous system(CNS) disorders is hindered by the limited ability of the vertebrate CNS to regenerate injured cells, replace damaged myelin sheath, and re-establish functional neuronal connections. Of many CNS disorders including multiple sclerosis, stroke, and other trauma, spinal cord injury is one of the important diseases because of the direct association with the functional loss of the body. Previous studies suggest that substantial recovery of function might be achieved through regeneration of lost neuronal cells and remyelination of intact axon in spinal cord injury which is occurred frequently. As a therapeutic approach in spinal cord injury, recently, cell transplantation provides a potential solution for the treatment of spinal cord injury. This review describes the characteristics of spinal cord injury and presents some evidence supporting functional recovery after cell transplantation following spinal cord injury.

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Physiological and Genetic Mechanisms for Nitrogen-Use Efficiency in Maize

  • Mi, Guohua;Chen, Fanjun;Zhang, Fusuo
    • Journal of Crop Science and Biotechnology
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    • v.10 no.2
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    • pp.57-63
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    • 2007
  • Due to the strong influence of nitrogen(N) on plant productivity, a vast amount of N fertilizers is used to maximize crop yield. Over-use of N fertilizers leads to severe pollution of the environment, especially the aquatic ecosystem, as well as reducing farmer's income. Growing of N-efficient cultivars is an important prerequisite for integrated nutrient management strategies in both low- and high-input agriculture. Taking maize as a sample crop, this paper reviews the response of plants to low N stress, the physiological processes which may control N-use efficiency in low-N input conditions, and the genetic and molecular biological aspects of N-use efficiency. Since the harvest index(HI) of modern cultivars is quite high, further improvement of these cultivars to adapt to low N soils should aim to increase their capacity to accumulate N at low N levels. To achieve this goal, establishment and maintenance of a large root system during the growth period may be essential. To reduce the cost of N and carbon for root growth, a strong response of lateral root growth to nitrate-rich patches may be desired. Furthermore, a large proportion of N accumulated in roots at early growth stages should be remobilized for grain growth in the late filling stage to increase N-utilization efficiency. Some QTLs and genes related to maize yield as well as root traits have been identified. However, their significance in improving maize NUE at low N inputs in the field need to be elucidated.

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A whole genomic scan to detect selection signatures between Berkshire and Korean native pig breeds

  • Edea, Zewdu;Kim, Kwan-Suk
    • Journal of Animal Science and Technology
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    • v.56 no.7
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    • pp.23.1-23.7
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    • 2014
  • Background: Scanning of the genome for selection signatures between breeds may play important role in understanding the underlie causes for observable phenotypic variations. The discovery of high density single nucleotide polymorphisms (SNPs) provide a useful starting point to perform genome-wide scan in pig populations in order to identify loci/candidate genes underlie phenotypic variation in pig breeds and facilitate genetic improvement programs. However, prior to this study genomic region under selection in commercially selected Berkshire and Korean native pig breeds has never been detected using high density SNP markers. To this end, we have genotyped 45 animals using Porcine SNP60 chip to detect selection signatures in the genome of the two breeds by using the $F_{ST}$ approach. Results: In the comparison of Berkshire and KNP breeds using the FDIST approach, a total of 1108 outlier loci (3.48%) were significantly different from zero at 99% confidence level with 870 of the outlier SNPs displaying high level of genetic differentiation ($F_{ST}{\geq}0.490$). The identified candidate genes were involved in a wide array of biological processes and molecular functions. Results revealed that 19 candidate genes were enriched in phosphate metabolism (GO: 0006796; ADCK1, ACYP1, CAMK2D, CDK13, CDK13, ERN1, GALK2, INPP1; MAK, MAP2K5, MAP3K1, MAPK14, P14KB, PIK3C3, PRKC1, PTPRK, RNASEL, THBS1, BRAF, VRK1). We have identified a set of candidate genes under selection and have known to be involved in growth, size and pork quality (CART, AGL, CF7L2, MAP2K5, DLK1, GLI3, CA3 and MC3R), ear morphology and size (HMGA2 and SOX5) stress response (ATF2, MSRB3, TMTC3 and SCAF8) and immune response (HCST and RYR1). Conclusions: Some of the genes may be used to facilitate genetic improvement programs. Our results also provide insights for better understanding of the process and influence of breed development on the pattern of genetic variations.

The spectroscopic study of chemical reaction of laser-ablated aluminum-oxygen by high power laser (분광분석을 활용한 고에너지 레이저 환경에서의 알루미늄-산소 화학반응 연구)

  • Kim, Chang-hwan
    • Journal of the Korean Society for Aeronautical & Space Sciences
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    • v.44 no.9
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    • pp.789-795
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    • 2016
  • Laser-induced combustions and explosions generated by high laser irradiances were explored by Laser-Induced Breakdown Spectroscopy (LIBS). The laser used for target ablation is a Q-switched Nd:YAG laser with 7 ns pulse duration at wavelength of 1064 nm laser energies from 40 mJ to 2500 mJ ($6.88{\times}10^{10}-6.53{\times}10^{11}W/cm^2$). The plasma light source from aluminum detected by the echelle grating spectrometer and coupled to the gated ICCD(a resolution (${\lambda}/{\Delta}{\lambda}$) of 5000). This spectroscopic study has been investigated for obtaining both the atomic/molecular signals of aluminum-oxygen and the calculated ambient condition such as plasma temperature and electron density. The essence of the paper is observing specific electron density ratio which can support the processes of chemical reaction and combustion between ablated aluminum plume and oxygen from air by inducing high laser energy.

Netrin-1 Specifically Enhances Cell Spreading on Fibronectin in Human Glioblastoma Cells

  • Lee, Hyun-Kyoung;Seo, In-Ae;Shin, Yoon-Kyung;Lee, Sang-Hwa;Seo, Su-Young;Suh, Duk-Joon;Park, Hwan-Tae
    • The Korean Journal of Physiology and Pharmacology
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    • v.12 no.5
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    • pp.225-230
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    • 2008
  • Netrins are secreted molecules and involved in axon guidance, cell migration and tumor development. Recent studies revealed that netrins perform novel functions in such processes as epithelial development and angiogenesis without operating through the classical netrin receptors, DCC (Deleted in Colorectal Cancer) and Unc5h. In the present study, we investigated the roles of netrin-1 and its receptors in cell spreading of human glioblastoma cells, and found that netrin-1 haptotactically enhanced fibronectin-induced cell spreading and focal adhesion formation in U373 glioblastoma cells. Netrin-1 binding to the U373 cell membrane was blocked by an antibody against ${\alpha}v$ integrin subunit, but not by an anti-DCC or anti-Unc5h antibody. In addition, enhancement of the fibronectin response by netrin-1 was abrogated by a function blocking antibody against integrin ${\alpha}v{\beta}3$. Since the ${\alpha}v$ subunit of the integrin family plays an important role in the pathophysiological aspects of cell migration, including tumor angiogenesis and metastasis, our data provide important insight into the molecular mechanism of netrin function.

Comparative Analyses of Tomato yellow leaf curl virus C4 Protein-Interacting Host Proteins in Healthy and Infected Tomato Tissues

  • Kim, Namgyu;Kim, Jinnyun;Bang, Bongjun;Kim, Inyoung;Lee, Hyun-Hee;Park, Jungwook;Seo, Young-Su
    • The Plant Pathology Journal
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    • v.32 no.5
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    • pp.377-387
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    • 2016
  • Tomato yellow leaf curl virus (TYLCV), a member of the genus Begomovirus, is one of the most important viruses of cultivated tomatoes worldwide, mainly causing yellowing and curling of leaves with stunting in plants. TYLCV causes severe problems in sub-tropical and tropical countries, as well as in Korea. However, the mechanism of TYLCV infection remains unclear, although the function of each viral component has been identified. TYLCV C4 codes for a small protein involved in various cellular functions, including symptom determination, gene silencing, viral movement, and induction of the plant defense response. In this study, through yeast-two hybrid screenings, we identified TYLCV C4-interacting host proteins from both healthy and symptom-exhibiting tomato tissues, to determine the role of TYLCV C4 proteins in the infection processes. Comparative analyses of 28 proteins from healthy tissues and 36 from infected tissues showing interactions with TYLCV C4 indicated that TYLCV C4 mainly interacts with host proteins involved in translation, ubiquitination, and plant defense, and most interacting proteins differed between the two tissues but belong to similar molecular functional categories. Four proteins-two ribosomal proteins, S-adenosyl-L-homocysteine hydrolase, and 14-3-3 family protein-were detected in both tissues. Furthermore, the identified proteins in symptom-exhibiting tissues showed greater involvement in plant defenses. Some are key regulators, such as receptor-like kinases and pathogenesis-related proteins, of plant defenses. Thus, TYLCV C4 may contribute to the suppression of host defense during TYLCV infection and be involved in ubiquitination for viral infection.

Characterization of Hibiscus Latent Fort Pierce Virus-Derived siRNAs in Infected Hibiscus rosa-sinensis in China

  • Lan, Han-hong;Lu, Luan-mei
    • The Plant Pathology Journal
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    • v.36 no.6
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    • pp.618-627
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    • 2020
  • Although limited progress have been made about pathogen system of Hibiscus rosa-sinensis and Hibiscus latent Fort Pierce virus (HLFPV), interaction between plant host and pathogen remain largely unknown, which led to deficiency of effective measures to control disease of hibiscus plants caused by HLFPV. In this study, infection of HLFPV in Hibiscus rosa-sinensis was firstly confirmed for the first time by traditional electron microscopy, modern reverse transcription polymerase chain reaction and RNA-seq methods in China (HLFPV-Ch). Sequence properties analyzing suggested that the full-length sequences (6,465 nt) of HLFPV-Ch had a high sequence identity and a similar genomic structure with other tobamoviruses. It includes a 5'-terminal untranslated region (UTR), followed by four open reading frames encoding for a 128.5-kDa replicase, a 186.5-kDa polymerase, a 31-kDa movement protein, 17.6-kDa coat protein, and the last a 3'-terminal UTR. Furthermore, HLFPV-Ch-derived virus-derived siRNAs (vsiRNAs) ant its putative target genes, reported also for the first time, were identified and characterized from disease Hibiscus rosa-sinensis through sRNA-seq and Patmatch server to investigate the interaction in this pathogen systems. HLFPV-Ch-derived vsiRNAs demonstrated several general and specific characteristics. Gene Ontology classification revealed predicted target genes by vsiRNAs are involved in abroad range of cellular component, molecular function and biological processes. Taken together, for first time, our results certified the HLFPV infection in China and provide an insight into interaction between HLFPV and Hibiscus rosa-sinensis.