RESUMO
In general, it is known that extreme climatic conditions such as El Niño and positive Indian Ocean Dipole (IOD+) cause prolonged drought in Indonesia's tropical peatlands so that groundwater levels (GWL) drop and peat is prone to fire. However, 27 years of GWL measurements in Central Kalimantan peat forests show the opposite condition, where the lowest GWL occurs several weeks before El Niño and after IOD+ reaches its peaks. We show that the dropped sea surface temperature anomaly induced by anomalously easterly winds along the southern Java-Sumatra occurs several weeks before the GWL drop to the lowest value. Local rainfall decreased, and GWL dropped sharply by 1.0 to 1.5 m, during the super El Niño events in 1997/98 and 2015, as well as remarkable events of IOD+ in 2019. It is suggested that the tropical peatland ecohydrological system (represented by the GWL), El Niño Southern Oscillation (ENSO), and IOD+ are teleconnected. Hence, monitoring GWL variability of peatland over the IMC is a possibility an alert for extreme climate events associated with El Niño and/or moderate IOD+.
Assuntos
El Niño Oscilação Sul , Água Subterrânea , Indonésia , Estações do Ano , Oceano Índico , SoloRESUMO
Splicing precursor messenger RNA (pre-mRNA) is a critical step to produce physiologically functional protein. Splicing failure not only gives rise to dysfunctional proteins but also generates abnormal protein function, which causes several diseases. Several pre-mRNA splicing factors are reported to regulate mitosis directly at mitotic structures and/or indirectly through controlling the pre-mRNA splicing for mitotic proteins. In this study, we described the mitotic functions of SF3B14, a component of the spliceosomal U2 small nuclear ribonucleoprotein (snRNP), which we identified as a candidate involved in mitosis based on the large-scale RNA interference (RNAi) screen of the nucleolar proteome database. We observed that SF3B14 depletion caused prolonged mitosis and several mitotic defects, such as monopolar spindle and chromosome misalignment during metaphase. Although SF3B14 was found in the nucleolar proteome database, our immunofluorescent stainings demonstrated that SF3B14 was predominantly localized in the nucleoplasm and excluded from the nucleolus during interphase. In addition, SF3B14 did not colocalize with specific mitotic structures during mitosis, which is not in line with its direct mitotic function. Notably, we found that the SF3B14 depletion reduced protein levels of TUBGCP6, required for centrosome regulation, and increased the unspliced/spliced ratio of its mRNA. Taken together, we propose that the pre-mRNA of TUBGCP6 is one of the targets for SF3B14 splicing through which SF3B14 controls mitotic chromosome behavior.
Assuntos
Centrossomo/metabolismo , Proteínas Associadas aos Microtúbulos/genética , Fosfoproteínas/metabolismo , Precursores de RNA/metabolismo , Splicing de RNA/genética , Cromossomos Humanos/metabolismo , Células HeLa , Humanos , Proteínas Associadas aos Microtúbulos/metabolismo , Mitose , Precursores de RNA/genética , Fatores de Processamento de RNA/metabolismoRESUMO
The nucleolus is a membrane-less nuclear structure that disassembles when cells undergo mitosis. During mitosis, nucleolar factors are thus released from the nucleolus and dynamically change their subcellular localization; however, their functions remain largely uncharacterised. Here, we found that a nucleolar factor called nucleolar protein 11 (NOL11) forms a protein complex with two tryptophan-aspartic acid (WD) repeat proteins named WD-repeat protein 43 (WDR43) and Cirhin in mitotic cells. This complex, referred to here as the NWC (NOL11-WDR43-Cirhin) complex, exists in nucleoli during interphase and translocates to the periphery of mitotic chromosomes, i.e., perichromosomal regions. During mitotic progression, both the congression of chromosomes to the metaphase plate and sister chromatid cohesion are impaired in the absence of the NWC complex, as it is required for the centromeric enrichment of Aurora B and the associating phosphorylation of histone H3 at threonine 3. These results reveal the characteristics of a novel protein complex consisting of nucleolar proteins, which is required for regulating kinetochores and centromeres to ensure faithful chromosome segregation.
Assuntos
Aurora Quinase B/metabolismo , Segregação de Cromossomos , Mitose , Proteínas Nucleares/metabolismo , Células HeLa , Humanos , Proteínas Nucleares/genética , Ligação ProteicaRESUMO
Protein arginine methyltransferase PRMT5 synthesizes the symmetric dimethylarginine in nuclear and cytoplasmic proteins such as histone H2A, H4 and several non-histone proteins that are required for a variety of biological processes. Currently, two splice variants (v1 and v2) of murine PRMT5 have been deposited in the NCBI sequence database, in which PRMT5-v1 and -v2 contain different 33 and 16â¯amino acids at the N-terminal sequences, respectively. Here we showed that murine PRMT5-v1 is stable, but PRMT5-v2 is constantly degraded through both the ubiquitin proteasome system (UPS) and the autophagic-lysosomal pathway (ALP) in an N-terminal sequence-dependent manner. Furthermore, inhibition of UPS and ALP elevated the stability of PRMT5-v2 that made it localized in the nucleus and the cytoplasm. In addition, PRMT5-v2 exhibited the enzyme activity to catalyze histone H2A and H4 methylation. Notably, we found that the heat shock protein (Hsp) 70 specially recognizes the N-terminal sequence of PRMT5-v2 and the carboxyl terminus of Hsp70-interacting protein (CHIP) is required for poly-ubiquitination and the degradation of PRMT5-v2. These results suggest that Hsp70/CHIP chaperone-mediated protein degradation system is crucial in the regulation of PRMT5-v2 turnover, which has the potential to balance the symmetrical arginine dimethylation in cells.
Assuntos
Proteínas Adaptadoras de Transdução de Sinal/metabolismo , Proteínas de Choque Térmico HSP70/metabolismo , Proteína-Arginina N-Metiltransferases/metabolismo , Ubiquitina-Proteína Ligases/metabolismo , Animais , Células Cultivadas , Feminino , Células HEK293 , Humanos , Masculino , Camundongos , Camundongos Endogâmicos C57BL , Proteína-Arginina N-Metiltransferases/genéticaRESUMO
Immiscible composite materials with controlled phase-separated structures are important in areas ranging from catalysis to battery. We succeeded in controlling the phase-separated structures of immiscible blends of polystyrene (PS) and two ionic liquids (ILs), namely, N, N-diethyl- N-(2-methoxyethyl)- N-methylammonium bis(trifluoromethylsulfonyl)imide (DEME-TFSI) and 1-ethyl-3-methylimidazolium bis(trifluoromethylsulfonyl)imide, by adding precisely designed concentrated polymer brush-grafted (CPB-grafted) silica nanoparticles (CPB-SiPs) prepared by surface-initiated atom-transfer radical polymerization. We discuss relationships between chemical species and molecular weights of the CPB and phase-separated structures. When the CPB was composed of a PS homopolymer of an appropriate molecular weight, the IL phase formed a continuous structure and a quasi-solid-blended film was successfully fabricated because the CPB-SiPs were adsorbed at the PS/IL interface and prevented macroscopic phase separation. We propose that CPB-SiP adsorption and the fabrication of quasi-solid films are governed by the degree of penetration of the matrix PS chains into the CPB and deformability of the CPB-SiPs. We found that the DEME-TFSI domain size can be controlled by the CPB-SiP content and that only 1 wt % of the CPB-SiPs was needed to fabricate a quasi-solid film. In addition, we investigated the ionic properties of the quasi-solid PS/DEME-TFSI-blended film. Owing to continuous ion channels composed only of DEME-TFSI, the film exhibited an ionic conductivity of 0.1 mS/cm, which is relatively high compared to previously reported quasi-solid electrolytes. Finally, we demonstrated that an electric double-layer capacitor fabricated using this film as the electrolyte exhibited high charge/discharge cycling stability and reversibility.
RESUMO
The nucleolus is a dynamic nuclear body that has been demonstrated to disassemble at the onset of mitosis; the relationship between cell cycle progression and nucleolar integrity, however, remains poorly understood. We studied the role of nucleolar proteins in mitosis by performing a global analysis using small interfering RNAs specific to nucleolar proteins; we focused on nucleolar protein 11 (NOL11), with currently unknown mitotic functions. Depletion of NOL11 delayed entry into the mitotic phase owing to increased inhibitory phosphorylation of cyclin-dependent kinase 1 (Cdk1) and aberrant accumulation of Wee1, a kinase that phosphorylates and inhibits Cdk1. In addition to effects on overall mitotic phenotypes, NOL11 depletion reduced ribosomal RNA (rRNA) levels and caused nucleolar disruption during interphase. Notably, mitotic phenotypes found in NOL11-depleted cells were recapitulated when nucleolar disruption was induced by depletion of rRNA transcription factors or treatment with actinomycin D. Furthermore, delayed entry into the mitotic phase, caused by the depletion of pre-rRNA transcription factors, was attributable to nucleolar disruption rather than to G2/M checkpoint activation or reduced protein synthesis. Our findings therefore suggest that maintenance of nucleolar integrity during interphase is essential for proper cell cycle progression to mitosis via the regulation of Wee1 and Cdk1.
Assuntos
Proteína Quinase CDC2/metabolismo , Nucléolo Celular/metabolismo , Interfase , Mitose , Ciclo Celular , Proteínas de Ciclo Celular/metabolismo , Ativação Enzimática , Células HeLa , Humanos , Proteínas Nucleares/metabolismo , Transporte Proteico , Proteínas Tirosina Quinases/metabolismoRESUMO
Inflammatory bowel disease (IBD), including ulcerative colitis and Crohn's disease, is a chronic intestinal inflammatory condition initiated by integrins-mediated leukocyte adhesion to the activated colonic microvascular endothelium. Calreticulin (CRT), a calcium-binding chaperone, is known as a partner in the activation of integrin α subunits (ITGAs). The relationship between their interaction and the pathogenesis of IBD is largely unknown. Here we show that a small molecule, orally active ER-464195-01, inhibits the CRT binding to ITGAs, which suppresses the adhesiveness of both T cells and neutrophils. Transcriptome analysis on colon samples from dextran sodium sulfate-induced colitis mice reveals that the increased expression of pro-inflammatory genes is downregulated by ER-464195-01. Its prophylactic and therapeutic administration to IBD mouse models ameliorates the severity of their diseases. We propose that leukocytes infiltration via the binding of CRT to ITGAs is necessary for the onset and development of the colitis and the inhibition of this interaction may be a novel therapeutic strategy for the treatment of IBD.
Assuntos
Anti-Inflamatórios/farmacologia , Calreticulina/imunologia , Colite Ulcerativa/imunologia , Cicloexanos/farmacologia , Cadeias alfa de Integrinas/imunologia , Piperazinas/farmacologia , Animais , Anti-Inflamatórios/uso terapêutico , Calreticulina/antagonistas & inibidores , Colite Ulcerativa/induzido quimicamente , Colite Ulcerativa/tratamento farmacológico , Colo/citologia , Colo/imunologia , Colo/patologia , Cicloexanos/uso terapêutico , Sulfato de Dextrana/toxicidade , Modelos Animais de Doenças , Feminino , Voluntários Saudáveis , Humanos , Cadeias alfa de Integrinas/metabolismo , Células Jurkat , Masculino , Camundongos , Camundongos Endogâmicos BALB C , Camundongos SCID , Infiltração de Neutrófilos/efeitos dos fármacos , Infiltração de Neutrófilos/imunologia , Neutrófilos/efeitos dos fármacos , Neutrófilos/imunologia , Piperazinas/uso terapêutico , Ligação Proteica , Linfócitos T/efeitos dos fármacos , Linfócitos T/imunologiaRESUMO
The perichromosomal layer (PCL) is a structure that surrounds mitotic chromosomes, found in both animal and plant cells. It comprises various proteins and RNAs, mainly derived from the nucleolus. Several functions for the PCL have been suggested; however, the mechanism of PCL organization during mitosis remains unclear. The localization of several nucleolar proteins to the PCL is reportedly dependent on pre-ribosomal RNAs and the marker of proliferation, Ki67, which is a major PCL-localized protein. Here we demonstrate that, although the removal of pre-ribosomal RNAs from the PCL causes PCL delocalization of several nucleolar proteins, it does not affect the localization of Ki67. Conversely, Ki67 depletion results in the dissociation of both pre-ribosomal RNAs and nucleolar proteins from the PCL, which indicates that Ki67 is required for the PCL accumulation of pre-ribosomal RNAs, to which several nucleolar proteins are associated. Given these findings, we propose a model for PCL organization that comprises three essential layers: the scaffolding protein Ki67, pre-ribosomal RNAs for linkage, and outer nucleolar proteins.
Assuntos
Nucléolo Celular/química , Antígeno Ki-67/análise , Proteínas Nucleares/análise , Precursores de RNA/análise , RNA Ribossômico/análise , Nucléolo Celular/metabolismo , Nucléolo Celular/ultraestrutura , Cromossomos/química , Cromossomos/metabolismo , Células HeLa , Humanos , Antígeno Ki-67/metabolismo , Mitose , Proteínas Nucleares/metabolismo , Precursores de RNA/metabolismo , RNA Ribossômico/metabolismoRESUMO
BACKGROUND: Asthma patients with fixed airflow limitation (FL) are theoretically classified into two phenotypes, that is, coexisting chronic obstructive pulmonary disease (COPD) and asthmatic airway remodeling. However, the precise percentages of such patients are not known. OBJECTIVE: To assess the prevalence of patients with both FL and COPD components in elderly asthma. METHODS: We evaluated patients by lung diffusion impairment and emphysematous findings in high-resolution computed tomography (HRCT) as candidates for COPD components, as a multicenter, cross-sectional survey. Asthma outpatients ≥ 50 years of age were enrolled from Tohoku University Hospital, Sendai, Japan, and four hospitals (Tohoku Medical and Pharmaceutical University Wakabayashi Hospital, Sendai, JAPAN; Wakayama Medical University Hospital, Kimiidera, Japan; Hiraka General Hospital, Yokote, Japan; Iwate Prefectural Isawa Hospital, Oshu, Japan) with pulmonary physicians from March 1, 2013 to November 30, 2014. RESULTS: The prevalence of patients with FEV1/FVC <70% was 31.0% of those in their 50s, 40.2% of those in their 60s and 61.9% of those in their 70s or older. The prevalence of those patients with lung diffusion impairment (i.e. the percent predicted values of diffusing capacity of the lung for carbon monoxide (DLco %predicted) <80%) or emphysematous findings in HRCT (i.e. the appearance of low attenuation area (LAA)) was 18.3% of those in their 50s, 13.8% of those in their 60s and 35.7% of those in their 70s or older. CONCLUSIONS: Nearly half of the patients with FL in elderly asthma show coexisting COPD components when assessed by DLco %predicted and LAA in HRCT.
Assuntos
Asma/epidemiologia , Asma/fisiopatologia , Doença Pulmonar Obstrutiva Crônica/epidemiologia , Doença Pulmonar Obstrutiva Crônica/fisiopatologia , Troca Gasosa Pulmonar/fisiologia , Idoso , Remodelação das Vias Aéreas/fisiologia , Asma/diagnóstico por imagem , Estudos Transversais , Feminino , Humanos , Japão/epidemiologia , Masculino , Pessoa de Meia-Idade , Doença Pulmonar Obstrutiva Crônica/diagnóstico por imagem , Testes de Função Respiratória , Fumar/epidemiologia , Tomografia Computadorizada por Raios X/métodosRESUMO
Asthma-chronic obstructive pulmonary disease (COPD) overlap syndrome (ACOS) was proposed by the science committees of both Global Initiative for Asthma (GINA) and Global Initiative for Chronic Obstructive Lung Disease (GOLD). However, the definition of ACOS has remained unclear all over the world, and the prevalence rate of ACOS is basically dependent on the patient's symptoms or the physician's opinion, based on questionnaire testing. In the current case report, we investigated the prevalence rate of COPD patients with high levels of fractional exhaled nitric oxide (FENO) or immunoglobulin E (IgE) as candidate markers of ACOS in COPD, as a multicenter, cross-sectional study. Outpatients with COPD were enrolled from Tohoku University Hospital, Sendai, Japan, and five hospitals (Tohoku University Hospital, Sendai, Japan; NTT East Tohoku Hospital, Sendai, Japan; Wakayama Medical University Hospital, Kimiidera, Japan; Hiraka General Hospital, Yokote, Japan; Iwate Prefectural Isawa Hospital, Oshu, Japan) with pulmonary physicians from March 1, 2013 to February 28, 2014. When they were estimated using 35 ppb as the cutoff value of FENO, the prevalence rate of ACOS was 16.3% in COPD. When estimated by both FENO and IgE, the high-FENO/high-IgE group was 7.8% in COPD. To the best of our knowledge, this study is the first to detect the prevalence rate of ACOS in COPD populations by using objective biomarkers. The results from the current study should be useful to identify the subgroup requiring early intervention by inhaled corticosteroids/long-acting beta agonist combination in COPD in order to improve the long-term management for ACOS.
Assuntos
Asma , Imunoglobulina E , Óxido Nítrico , Doença Pulmonar Obstrutiva Crônica , Eliminação Pulmonar , Idoso , Asma/diagnóstico , Asma/epidemiologia , Asma/metabolismo , Asma/fisiopatologia , Biomarcadores/análise , Biomarcadores/metabolismo , Testes Respiratórios/métodos , Estudos Transversais , Feminino , Humanos , Imunoglobulina E/análise , Imunoglobulina E/metabolismo , Japão/epidemiologia , Masculino , Óxido Nítrico/análise , Óxido Nítrico/metabolismo , Prevalência , Doença Pulmonar Obstrutiva Crônica/diagnóstico , Doença Pulmonar Obstrutiva Crônica/epidemiologia , Doença Pulmonar Obstrutiva Crônica/metabolismo , Doença Pulmonar Obstrutiva Crônica/fisiopatologia , Testes de Função Respiratória/métodos , SíndromeRESUMO
The nucleolus, whose primary function is ribosome biogenesis, plays an essential role in p53 activation. Ribosome biogenesis is inhibited in response to cellular stress and several nucleolar proteins translocate from the nucleolus to the nucleoplasm, where they activate p53. In this study, we analysed precisely how impaired ribosome biogenesis regulates the activation of p53 by depleting nucleolar factors involved in rRNA transcription or rRNA processing. Nucleolar RNA content decreased when rRNA transcription was inhibited. In parallel with the reduced levels of nucleolar RNA content, the nucleolar protein Myb-binding protein 1 A (MYBBP1A) translocated to the nucleoplasm and increased p53 acetylation. The acetylated p53 enhanced p21 and BAX expression and induced apoptosis. In contrast, when rRNA processing was inhibited, MYBBP1A remained in the nucleolus and nonacetylated p53 accumulated, causing cell cycle arrest at the G1 phase by inducing p21 but not BAX. We propose that the nucleolus functions as a stress sensor to modulate p53 protein levels and its acetylation status, determining cell fate between cell cycle arrest and apoptosis by regulating MYBBP1A translocation.
Assuntos
Apoptose/genética , Proteínas Nucleares/metabolismo , Proteínas de Transporte Nucleocitoplasmático/metabolismo , RNA Ribossômico/genética , Transcrição Gênica , Proteína Supressora de Tumor p53/metabolismo , Acetilação , Linhagem Celular Tumoral , Núcleo Celular/metabolismo , Proteínas de Ligação a DNA , Pontos de Checagem da Fase G1 do Ciclo Celular/genética , Humanos , Modelos Biológicos , Transporte Proteico , Interferência de RNA , Proteínas de Ligação a RNA , Proteínas Ribossômicas/metabolismo , Fatores de TranscriçãoAssuntos
Estudos Transversais , Estudos Multicêntricos como Assunto , Doença Pulmonar Obstrutiva Crônica/diagnóstico , Idoso , Idoso de 80 Anos ou mais , Progressão da Doença , Dispneia/diagnóstico , Dispneia/fisiopatologia , Feminino , Volume Expiratório Forçado , Humanos , Masculino , Pessoa de Meia-Idade , Prognóstico , Doença Pulmonar Obstrutiva Crônica/fisiopatologia , Risco , Índice de Gravidade de Doença , Inquéritos e Questionários , Capacidade VitalRESUMO
Various cellular stresses activate autophagy, which is involved in lysosomal degradation of cytoplasmic materials for maintaining nutrient homeostasis and eliminating harmful components. Here, we show that RNA polymerase I (Pol I) transcription inhibition induces nucleolar disruption and autophagy. Treatment with autophagy inhibitors or siRNA specific for autophagy-related (ATG) proteins inhibited autophagy but not nucleolar disruption induced by Pol I transcription inhibition, which suggested that nucleolar disruption was upstream of autophagy. Furthermore, treatment with siRNA specific for nucleolar protein nucleophosmin (NPM) inhibited this type of autophagy. This showed that NPM was involved in autophagy when the nucleolus was disrupted by Pol I inhibition. In contrast, NPM was not required for canonical autophagy induced by nutrient starvation, as it was not accompanied by nucleolar disruption. Thus, our results revealed that, in addition to canonical autophagy, there may be NPM-dependent autophagy associated with nucleolar disruption.
Assuntos
Autofagia/genética , Proteínas Nucleares/genética , RNA Polimerase I/genética , Transcrição Gênica , Nucléolo Celular/genética , Humanos , Células MCF-7 , Proteínas Nucleares/antagonistas & inibidores , Região Organizadora do Nucléolo/genética , Nucleofosmina , RNA Polimerase I/antagonistas & inibidores , RNA Interferente PequenoRESUMO
The 5S ribonucleoprotein particle (RNP) complex, consisting of RPL11, RPL5, and 5S rRNA, is implicated in p53 regulation under ribotoxic stress. Here, we show that the 5S RNP contributes to p53 activation and promotes cellular senescence in response to oncogenic or replicative stress. Oncogenic stress accelerates rRNA transcription and replicative stress delays rRNA processing, resulting in RPL11 and RPL5 accumulation in the ribosome-free fraction, where they bind MDM2. Experimental upregulation of rRNA transcription or downregulation of rRNA processing, mimicking the nucleolus under oncogenic or replicative stress, respectively, also induces RPL11-mediated p53 activation and cellular senescence. We demonstrate that exogenous expression of certain rRNA-processing factors rescues the processing defect, attenuates p53 accumulation, and increases replicative lifespan. To summarize, the nucleolar-5S RNP-p53 pathway functions as a senescence inducer in response to oncogenic and replicative stresses.
Assuntos
Senescência Celular , Ribossomos/metabolismo , Proteína Supressora de Tumor p53/metabolismo , Animais , Proteínas de Ciclo Celular/antagonistas & inibidores , Proteínas de Ciclo Celular/genética , Proteínas de Ciclo Celular/metabolismo , Nucléolo Celular/metabolismo , Células Cultivadas , Humanos , Células MCF-7 , Camundongos , Proteínas Nucleares/antagonistas & inibidores , Proteínas Nucleares/genética , Proteínas Nucleares/metabolismo , Proteínas Proto-Oncogênicas c-mdm2/genética , Proteínas Proto-Oncogênicas c-mdm2/metabolismo , Interferência de RNA , RNA Ribossômico 5S/metabolismo , RNA Interferente Pequeno/metabolismo , Proteínas de Ligação a RNA , Proteínas Ribossômicas/antagonistas & inibidores , Proteínas Ribossômicas/genética , Proteínas Ribossômicas/metabolismo , Ativação Transcricional , Regulação para CimaRESUMO
Cancer stem cells (CSCs) have several distinctive characteristics, including high metastatic potential, tumor-initiating potential, and properties that resemble normal stem cells such as self-renewal, differentiation, and drug efflux. Because of these characteristics, CSC is regarded to be responsible for cancer progression and patient prognosis. In our previous study, we showed that a ubiquitin E3 ligase carboxyl terminus of Hsc70-interacting protein (CHIP) suppressed breast cancer malignancy. Moreover, a recent clinical study reported that CHIP expression levels were associated with favorable prognostic parameters of patients with breast cancer. Here we show that CHIP suppresses CSC properties in a population of breast cancer cells. CHIP depletion resulted in an increased proportion of CSCs among breast cancers when using several assays to assess CSC properties. From our results, we propose that inhibition of CSC properties may be one of the functions of CHIP as a suppressor of cancer progression.
Assuntos
Neoplasias da Mama/metabolismo , Neoplasias da Mama/patologia , Células-Tronco Neoplásicas/metabolismo , Células-Tronco Neoplásicas/patologia , Ubiquitina-Proteína Ligases/metabolismo , Diferenciação Celular , Proliferação de Células , Feminino , Humanos , Células MCF-7RESUMO
An imaging plate (IP) system was used as an effective detector for direct measurement of radioactive surface contamination. The IP system displayed images designating the locations and extent of fixed surface contamination of uranyl acetate. The amount of radioactive waste produced during decontamination was reduced because the contaminated spots could be isolated; furthermore, creation of radioactive dust during removal of contamination was prevented because the contaminated spots could be removed without being pulverized. The images were used in efficiently and safely isolating the location of fixed surface contamination. The IP system surface contamination detection limit for uranyl acetate was 2.5 × 10 Bq cm, a value much lower than the surface contamination limit and the clearance level.
Assuntos
Monitoramento de Radiação/instrumentação , Monitoramento de Radiação/métodos , Calibragem , Segurança , Propriedades de SuperfícieRESUMO
Responding to various stimuli is indispensable for the maintenance of homeostasis. The downregulation of ribosomal RNA (rRNA) transcription is one of the mechanisms involved in the response to stimuli by various cellular processes, such as cell cycle arrest and apoptosis. Cell differentiation is caused by intra- and extracellular stimuli and is associated with the downregulation of rRNA transcription as well as reduced cell growth. The downregulation of rRNA transcription during differentiation is considered to contribute to reduced cell growth. However, the downregulation of rRNA transcription can induce various cellular processes; therefore, it may positively regulate cell differentiation. To test this possibility, we specifically downregulated rRNA transcription using actinomycin D or a siRNA for Pol I-specific transcription factor IA (TIF-IA) in HL-60 and THP-1 cells, both of which have differentiation potential. The inhibition of rRNA transcription induced cell differentiation in both cell lines, which was demonstrated by the expression of the common differentiation marker CD11b. Furthermore, TIF-IA knockdown in an ex vivo culture of mouse hematopoietic stem cells increased the percentage of myeloid cells and reduced the percentage of immature cells. We also evaluated whether differentiation was induced via the inhibition of cell cycle progression because rRNA transcription is tightly coupled to cell growth. We found that cell cycle arrest without affecting rRNA transcription did not induce differentiation. To the best of our knowledge, our results demonstrate the first time that the downregulation of rRNA levels could be a trigger for the induction of differentiation in mammalian cells. Furthermore, this phenomenon was not simply a reflection of cell cycle arrest. Our results provide a novel insight into the relationship between rRNA transcription and cell differentiation.
Assuntos
Diferenciação Celular/genética , Regulação para Baixo/genética , RNA Ribossômico/genética , Transcrição Gênica/genética , Animais , Ciclo Celular/genética , Pontos de Checagem do Ciclo Celular/genética , Linhagem Celular , Linhagem Celular Tumoral , Células HL-60 , Humanos , Masculino , Camundongos , Camundongos Endogâmicos C57BL , Células Mieloides/metabolismo , Proteínas Pol1 do Complexo de Iniciação de Transcrição/genética , RNA Polimerase I/genética , RNA Interferente Pequeno/genética , Fatores de TranscriçãoRESUMO
Ribosome biosynthesis is a major intracellular energy-consuming process. We previously identified a nucleolar factor, nucleomethylin (NML), which regulates intracellular energy consumption by limiting rRNA transcription. Here, we show that, in livers of obese mice, the recruitment of NML to rRNA gene loci is increased to repress rRNA transcription. To clarify the relationship between obesity and rRNA transcription, we generated NML-null (NML-KO) mice. NML-KO mice show elevated rRNA level, reduced ATP concentration, and reduced lipid accumulation in the liver. Furthermore, in high-fat-diet (HFD)-fed NML-KO mice, hepatic rRNA levels are not decreased. Both weight gain and fat accumulation in HFD-fed NML-KO mice are significantly lower than those in HFD-fed wild-type mice. These findings indicate that rRNA transcriptional activation promotes hepatic energy consumption, which alters hepatic lipid metabolism. Namely, hepatic rRNA transcriptional repression by HFD feeding is essential for energy storage.
Assuntos
Dieta Hiperlipídica , Fígado/metabolismo , RNA Ribossômico/metabolismo , Trifosfato de Adenosina/metabolismo , Animais , Metabolismo Energético , Ácidos Graxos/biossíntese , Expressão Gênica , Metabolismo dos Lipídeos/genética , Fígado/diagnóstico por imagem , Camundongos , Camundongos Endogâmicos C57BL , Camundongos Knockout , Camundongos Obesos , Proteínas Nucleares/deficiência , Proteínas Nucleares/genética , Proteínas Nucleares/metabolismo , Obesidade/etiologia , Obesidade/metabolismo , Obesidade/patologia , RNA Ribossômico/genética , Sirtuína 1/metabolismo , Tomografia Computadorizada por Raios X , Transcrição GênicaRESUMO
Adrenoleukodystrophy (ALD) is a genetic disorder with demyelination of the central nervous system and adrenal insufficiency. A 24-year-old man with ALD was scheduled for dental treatment under general anesthesia. He was diagnosted as having ALD at the age of 5. Past medical history included recurrent cervical cellulitis, adrenal insufficiency, mental retardation, muscle weakness and seizure disorder. General anesthesia was induced using betamethasone as a steroid cover, sevoflurane and nitrous oxide-oxygen and maintained with sevoflurane and nitrous oxide-oxygen. Nasal intubation was performed without using a muscle relaxant. Patients with ALD cannot metabolize very long chain fatty acid, so we did not use propofol containing long chain fatty acid. Operation and anesthesia were uneventful. There were no complications during and after anesthesia.
Assuntos
Adrenoleucodistrofia , Anestesia Geral/métodos , Assistência Odontológica para a Pessoa com Deficiência/métodos , Membro 1 da Subfamília D de Transportadores de Cassetes de Ligação de ATP , Transportadores de Cassetes de Ligação de ATP/genética , Transportadores de Cassetes de Ligação de ATP/fisiologia , Hiperfunção Adrenocortical , Adrenoleucodistrofia/etiologia , Adulto , Ácidos Graxos Insaturados/metabolismo , Doenças Desmielinizantes Hereditárias do Sistema Nervoso Central , Humanos , Intubação Intratraqueal/métodos , Masculino , Éteres Metílicos , Óxido Nitroso , Sevoflurano , Adulto JovemRESUMO
Tetramerization of p53 is crucial to exert its biological activity, and nucleolar disruption is sufficient to activate p53. We previously demonstrated that nucleolar stress induces translocation of the nucleolar protein MYBBP1A from the nucleolus to the nucleoplasm and enhances p53 activity. However, whether and how MYBBP1A regulates p53 tetramerization in response to nucleolar stress remain unclear. In this study, we demonstrated that MYBBP1A enhances p53 tetramerization, followed by acetylation under nucleolar stress. We found that MYBBP1A has two regions that directly bind to lysine residues of the p53 C-terminal regulatory domain. MYBBP1A formed a self-assembled complex that provided a molecular platform for p53 tetramerization and enhanced p300-mediated acetylation of the p53 tetramer. Moreover, our results show that MYBBP1A functions to enhance p53 tetramerization that is necessary for p53 activation, followed by cell death with actinomycin D treatment. Thus, we suggest that MYBBP1A plays a pivotal role in the cellular stress response.