Your browser doesn't support javascript.
loading
Mostrar: 20 | 50 | 100
Resultados 1 - 20 de 48
Filtrar
1.
Antonie Van Leeuwenhoek ; 105(1): 191-7, 2014 Jan.
Artigo em Inglês | MEDLINE | ID: mdl-24287870

RESUMO

A Gram-stain negative, strictly aerobic, non-motile, non-spore-forming, and rod-shaped bacterial strain designated FW-3(T) was isolated from fresh water and its taxonomic position was investigated by using a polyphasic approach. Strain FW-3(T) was found to grow at 10-37 °C and at pH 7.0 in the absence of NaCl on nutrient agar. On the basis of 16S rRNA gene sequence similarity, strain FW-3(T) was shown to belong to the family Acetobacteraceae and to be related to Roseomonas lacus TH-G33(T) (97.2 % sequence similarity) and Roseomonas terrae DS-48(T) (96.4 %). The G+C content of the genomic DNA was determined to be 68.0 %. The major menaquinone was determined to be Q-10 and the major fatty acids were identified as summed feature 7 (comprising C18:1 ω9c/ω12t/ω7c as defined by the MIDI system; 55.4 %), and C18:1 2OH (29.8 %). DNA and chemotaxonomic data supported the affiliation of strain FW-3(T) to the genus Roseomonas. Strain FW-3(T) could be differentiated genotypically and phenotypically from the recognized species of the genus Roseomonas. The novel isolate therefore represents a novel species, for which the name Roseomonas sediminicola sp. nov. is proposed, with the type strain FW-3(T) (=KACC 16616(T) = JCM 18210(T)).


Assuntos
Água Doce/microbiologia , Methylobacteriaceae/classificação , Methylobacteriaceae/isolamento & purificação , DNA Bacteriano/genética , DNA Ribossômico/genética , Ácidos Graxos/química , Ácidos Graxos/metabolismo , Methylobacteriaceae/genética , Methylobacteriaceae/metabolismo , Dados de Sequência Molecular , Filogenia , RNA Ribossômico 16S/genética
2.
Int J Syst Evol Microbiol ; 63(Pt 6): 1942-1946, 2013 Jun.
Artigo em Inglês | MEDLINE | ID: mdl-23024142

RESUMO

A strictly anaerobic, Gram-negative, non-spore-forming bacterium, designated GH1(T), was isolated from the rumen of Korean native cattle (HanWoo). Cells were straight to slightly curved rods (2.0-4.5 µm long) and were motile by peritrichous flagella. The isolate grew at 30-45 °C (optimum 40 °C), at pH 5.5-6.5 (optimum pH 6.0) and with up to 3.5% (w/v) NaCl. Strain GH1(T) produced acid from d-glucose, d-ribose and d-xylose, with butyric acid being the major end product. The genomic DNA G+C content was 54.6 mol%. Based on comparative 16S rRNA gene sequence analysis, strain GH1(T) was most closely related to Oscillibacter valericigenes Sjm18-20(T) (97.3% 16S rRNA gene sequence similarity). DNA-DNA hybridization between strain GH1(T) and O. valericigenes DSM 18026(T) showed 24% reassociation. The major fatty acids were iso-C13:0 (13.0%), iso-C15:0 (17.6%), anteiso-C15:0 (8.4%) and C14:0 (4.1%), and the cellular fatty acid methyl esters as dimethylacetals (DMAs) were C16:0 DMA (17.8%), iso-C15:0 DMA (15.2%) and C14:0 DMA (4.52%). The cell-wall peptidoglycan of strain GH1(T) contained meso-diaminopimelic acid and the major cell-wall sugar was galactose. Based on 16S rRNA gene sequence similarity, phylogenetic analysis, DNA G+C content, DNA-DNA relatedness and distinct phenotypic characteristics, strain GH1(T) is classified in the genus Oscillibacter as a member of a novel species, for which the name Oscillibacter ruminantium sp. nov. is proposed. The type strain is GH1(T) (=KCTC 15176(T)=NBRC 108824(T)=JCM 18333(T)).


Assuntos
Bovinos/microbiologia , Bactérias Gram-Negativas/classificação , Filogenia , Rúmen/microbiologia , Animais , Técnicas de Tipagem Bacteriana , Composição de Bases , DNA Bacteriano/genética , Ácidos Graxos/análise , Bactérias Gram-Negativas/genética , Bactérias Gram-Negativas/isolamento & purificação , Dados de Sequência Molecular , Hibridização de Ácido Nucleico , Peptidoglicano/análise , RNA Ribossômico 16S/genética , República da Coreia , Análise de Sequência de DNA
3.
Int J Syst Evol Microbiol ; 63(Pt 11): 4196-4201, 2013 Nov.
Artigo em Inglês | MEDLINE | ID: mdl-23771627

RESUMO

Three strictly anaerobic, methanogenic strains JH1(T), JH4 and JH8 were isolated from rumen of the Korean native cattle (HanWoo; Bos taurus coreanae) in South Korea. The colonies were circular, opaque, and slightly yellowish. Phylogenetic analyses of 16S rRNA gene and mcrA (encoding α subunit of methyl-coenzyme M reductase) sequences confirmed the affiliation of the novel strains with the Methanobacteriales, and Methanobrevibacter wolinii SH(T) was the most closely related species. The 16S rRNA gene and mcrA sequence similarities between strains JH1(T), JH4 and JH8 and M. wolinii SH(T) were 96.2 and 89.0 % respectively, and DNA-DNA hybridization of the isolates and M. wolinii DSM 11976(T) showed a 20 % reassociation. Strain JH1(T) exhibited 92 % DNA-DNA relatedness with strains JH4 and JH8, and their 16S rRNA gene and mcrA sequences were identical. Cells stained Gram-positive and were non-motile rods, 1.5-1.8 µm long and 0.6 µm wide. The strains were able to use H2/CO2 and formate. The optimum temperature and pH ranges for growth were 37-40 °C and pH 6.5-7.0. The DNA G+C content of strain JH1(T) was 28 mol%. Based on data from this study using a polyphasic approach, the three strains represent a novel species of genus Methanobrevibacter, for which the name Methanobrevibacter boviskoreani sp. nov. is proposed. The type strain is JH1(T) ( = KCTC 4102(T) = JCM 18376(T)).


Assuntos
Bovinos/microbiologia , Methanobrevibacter/classificação , Filogenia , Rúmen/microbiologia , Animais , Técnicas de Tipagem Bacteriana , Composição de Bases , Parede Celular/química , Methanobrevibacter/genética , Methanobrevibacter/isolamento & purificação , Dados de Sequência Molecular , Hibridização de Ácido Nucleico , Oxirredutases/genética , RNA Ribossômico 16S/genética , República da Coreia , Análise de Sequência de DNA
4.
Antonie Van Leeuwenhoek ; 103(6): 1385-93, 2013 Jun.
Artigo em Inglês | MEDLINE | ID: mdl-23553363

RESUMO

A Gram-positive, coccoid to rod-shaped, non-spore-forming bacterium, designated Gsoil 958(T), was isolated from soil of a ginseng field located in Pocheon province in South Korea. This bacterium was characterized in order to determine its taxonomic position by using a polyphasic approach. Strain Gsoil 958(T) was observed to grow well at 25-30 °C and at pH 7.0 on R2A and nutrient agar without NaCl supplementation. Strain Gsoil 958(T) was determined to have ß-glucosidase activity and the ability to transform ginsenoside Rb1 (one of the dominant active components of ginseng) to F2 via gypenoside XVII and Rd. On the basis of 16S rRNA gene sequence similarity, strain Gsoil 958(T) was shown to belong to the family Nocardioidaceae and related most closely to Nocardioides koreensis MSL-09(T) (97.6 % 16S rRNA gene sequence similarity), Nocardioides aquiterrae GW-9(T) (97.0 %), and Nocardioides sediminis MSL-01(T) (97.0 %). The sequence similarities with other validly named species within the genus Nocardioides were less than 96.8 %. Strain Gsoil 958(T) was characterized chemotaxonomically as having LL-2,6-diaminopimelic acid in the cell-wall peptidoglycan, MK-8(H4) as the predominant menaquinone, and iso-C16:0, iso-C16:1 H, iso-C14:0, iso-C15:0 were identified as the major fatty acids. The G + C content of genomic DNA was determined to be 70.8 mol %. The chemotaxonomic properties and phenotypic characteristics supported the affiliation of strain Gsoil 958(T) to the genus Nocardioides. The results of both physiological and biochemical tests allowed for differentiation of strain Gsoil 958(T) from the recognized Nocardioides species. Therefore, strain Gsoil 958(T) is considered to represent a novel species of the genus Nocardioides, for which the name Nocardioides panaciterrulae sp. nov. is proposed, with the type strain Gsoil 958(T) (KACC 14271(T) = KCTC 19471(T) = DSM 21350(T)).


Assuntos
Ginsenosídeos/metabolismo , Panax/microbiologia , Propionibacteriaceae/isolamento & purificação , Propionibacteriaceae/metabolismo , Microbiologia do Solo , Técnicas de Tipagem Bacteriana , Composição de Bases , Sequência de Bases , DNA Bacteriano/genética , Ácido Diaminopimélico/análise , Ácidos Graxos/análise , Dados de Sequência Molecular , Filogenia , Propionibacteriaceae/classificação , Propionibacteriaceae/genética , RNA Ribossômico 16S/genética , República da Coreia , Análise de Sequência de DNA , Vitamina K 2/análise , beta-Glucosidase/metabolismo
5.
J Bacteriol ; 194(22): 6362, 2012 Nov.
Artigo em Inglês | MEDLINE | ID: mdl-23105088

RESUMO

Oscillibacter ruminantium strain GH1 was isolated from the rumen of Korean native cattle (HanWoo; Bos taurus coreanae). Here, we present the 3.07-Mb draft genome of this strain, which could reveal the presence of certain fiber-specific glycoside hydrolases and butyric acid-producing genes.


Assuntos
Bactérias/classificação , Bactérias/genética , Bovinos/microbiologia , Genoma Bacteriano , Rúmen/microbiologia , Animais , Dados de Sequência Molecular , República da Coreia
6.
Materials (Basel) ; 15(24)2022 Dec 18.
Artigo em Inglês | MEDLINE | ID: mdl-36556862

RESUMO

Although artificial porous materials are useful for dissipating acoustic waves, they pose a major environmental threat as most are non-recyclable. Developing sustainable structural materials with the mechanical and energy-absorption properties required to replace artificial porous materials is currently a key challenge. Here, we report, for the first time, a novel microstructure using all-natural moss with a compressive strength of up to 2.35 GPa and a sound-absorption performance of up to 90%, depending on the additives, such as yogurt, starch, and beer. In addition, the moss-based microstructure was applied as graffiti to a three-dimensionally printed house model to demonstrate improved performance against the effects of sound. By incorporating energy-absorbing materials without harmful substances, the desired structure can be decorated with the graffiti method. This work could pave the way for attenuating sound-wave and impact noise by using graffiti work on structural composite materials.

7.
Mol Cancer Res ; 20(3): 412-424, 2022 Mar 01.
Artigo em Inglês | MEDLINE | ID: mdl-34728553

RESUMO

Loss of NF2 (merlin) has been suggested as a genetic cause of neurofibromatosis type 2 and malignant peripheral nerve sheath tumor (MPNST). Previously, we demonstrated that NF2 sustained TGFß receptor 2 (TßR2) expression and reduction or loss of NF2 activated non-canonical TGFß signaling, which reduced Raf kinase inhibitor protein (RKIP) expression via TßR1 kinase activity. Here, we show that a selective RKIP inducer (novel chemical, Nf18001) inhibits tumor growth and promotes schwannoma cell differentiation into mature Schwann cells under NF2-deficient conditions. In addition, Nf18001 is not cytotoxic to cells expressing NF2 and is not disturb canonical TGFß signaling. Moreover, the novel chemical induces expression of SOX10, a marker of differentiated Schwann cells, and promotes nuclear export and degradation of SOX2, a stem cell factor. Treatment with Nf18001 inhibited tumor growth in an allograft model with mouse schwannoma cells. These results strongly suggest that selective RKIP inducers could be useful for the treatment of neurofibromatosis type 2 as well as NF2-deficient MPNST. IMPLICATIONS: This study identifies that a selective RKIP inducer inhibits tumor growth and promotes schwannoma cell differentiation under NF2-deficient conditions by reducing SOX2 and increasing SOX10 expression.


Assuntos
Neurilemoma , Neurofibromatose 2 , Neurofibrossarcoma , Animais , Diferenciação Celular , Humanos , Camundongos , Neurilemoma/genética , Neurilemoma/metabolismo , Neurilemoma/patologia , Neurofibromatose 2/genética , Neurofibromina 2/genética , Neurofibromina 2/metabolismo , Proteína de Ligação a Fosfatidiletanolamina/metabolismo , Fatores de Transcrição SOXB1/genética , Fatores de Transcrição SOXB1/metabolismo , Fator de Crescimento Transformador beta/metabolismo
8.
Artigo em Inglês | MEDLINE | ID: mdl-33807417

RESUMO

In the present study we investigated the ability of the microalgal strain Parachlorella sp. AA1 to biologically uptake a radionuclide waste material. Batch experiments were conducted to investigate the biosorption of uranyl ions (U(VI)) in the 0.5-50.0 mg/L concentration range by strain AA1. The results showed that AA1 biomass could uptake U(VI). The highest removal efficiency and biosorption capacity (95.6%) occurred within 60 h at an initial U(VI) concentration of 20 mg/L. The optimum pH for biosorption was 9.0 at a temperature of 25 °C. X-ray absorption near edge structure analysis confirmed the presence of U(VI) in pellets of Parachlorella sp. AA1 cells. The biosorption methods investigated here may be useful in the treatment and disposal of nuclides and heavy metals in diverse wastewaters.


Assuntos
Clorófitas , Poluentes Químicos da Água , Adsorção , Biomassa , Concentração de Íons de Hidrogênio , Íons , Cinética
9.
Microorganisms ; 9(5)2021 May 20.
Artigo em Inglês | MEDLINE | ID: mdl-34065548

RESUMO

Agrobacterium tumefaciens is a pathogen of various plants which transfers its own DNA (T-DNA) to the host plants. It is used for producing genetically modified plants with this ability. To control T-DNA transfer to the right place, toxin-antitoxin (TA) systems of A. tumefaciens were used to control the target site of transfer without any unintentional targeting. Here, we describe a toxin-antitoxin system, Atu0939 (mazE-at) and Atu0940 (mazF-at), in the chromosome of Agrobacterium tumefaciens. The toxin in the TA system has 33.3% identity and 45.5% similarity with MazF in Escherichia coli. The expression of MazF-at caused cell growth inhibition, while cells with MazF-at co-expressed with MazE-at grew normally. In vivo and in vitro assays revealed that MazF-at inhibited protein synthesis by decreasing the cellular mRNA stability. Moreover, the catalytic residue of MazF-at was determined to be the 24th glutamic acid using site-directed mutagenesis. From the results, we concluded that MazF-at is a type II toxin-antitoxin system and a ribosome-independent endoribonuclease. Here, we characterized a TA system in A. tumefaciens whose understanding might help to find its physiological function and to develop further applications.

10.
Sci Rep ; 11(1): 9122, 2021 04 27.
Artigo em Inglês | MEDLINE | ID: mdl-33907225

RESUMO

Werner syndrome (WRN) is a rare progressive genetic disorder, caused by functional defects in WRN protein and RecQ4L DNA helicase. Acceleration of the aging process is initiated at puberty and the expected life span is approximately the late 50 s. However, a Wrn-deficient mouse model does not show premature aging phenotypes or a short life span, implying that aging processes differ greatly between humans and mice. Gene expression analysis of WRN cells reveals very similar results to gene expression analysis of Hutchinson Gilford progeria syndrome (HGPS) cells, suggesting that these human progeroid syndromes share a common pathological mechanism. Here we show that WRN cells also express progerin, an abnormal variant of the lamin A protein. In addition, we reveal that duplicated sequences of human WRN (hWRN) from exon 9 to exon 10, which differ from the sequence of mouse WRN (mWRN), are a natural inhibitor of progerin. Overexpression of hWRN reduced progerin expression and aging features in HGPS cells. Furthermore, the elimination of progerin by siRNA or a progerin-inhibitor (SLC-D011 also called progerinin) can ameliorate senescence phenotypes in WRN fibroblasts and cardiomyocytes, derived from WRN-iPSCs. These results suggest that progerin, which easily accumulates under WRN-deficient conditions, can lead to premature aging in WRN and that this effect can be prevented by SLC-D011.


Assuntos
Lamina Tipo A/metabolismo , Progéria/patologia , Helicase da Síndrome de Werner/metabolismo , Síndrome de Werner/genética , Adulto , Senilidade Prematura/genética , Animais , Linhagem Celular , Senescência Celular/efeitos dos fármacos , Criança , Modelos Animais de Doenças , Feminino , Fibroblastos/patologia , Expressão Gênica , Humanos , Masculino , Camundongos Mutantes , Progéria/genética , Isoformas de Proteínas , Síndrome de Werner/patologia , Helicase da Síndrome de Werner/genética
11.
Commun Biol ; 4(1): 1397, 2021 12 15.
Artigo em Inglês | MEDLINE | ID: mdl-34912047

RESUMO

Amyotrophic Lateral Sclerosis (ALS) is a fatal neurodegenerative disease characterized by selective death of motor neurons. Mutations in Cu, Zn-superoxide dismutase (SOD1) causing the gain of its toxic property are the major culprit of familial ALS (fALS). The abnormal SOD1 aggregation in the motor neurons has been suggested as the major pathological hallmark of ALS patients. However, the development of pharmacological interventions against SOD1 still needs further investigation. In this study, using ELISA-based chemical screening with wild and mutant SOD1 proteins, we screened a new small molecule, PRG-A01, which could block the misfolding/aggregation of SOD1 or TDP-43. The drug rescued the cell death induced by mutant SOD1 in human neuroblastoma cell line. Administration of PRG-A01 into the ALS model mouse resulted in significant improvement of muscle strength, motor neuron viability and mobility with extended lifespan. These results suggest that SOD1 misfolding/aggregation is a potent therapeutic target for SOD1 related ALS.


Assuntos
Esclerose Lateral Amiotrófica/genética , Neurônios Motores/fisiologia , Degeneração Neural/fisiopatologia , Dobramento de Proteína , Superóxido Dismutase-1/genética , Esclerose Lateral Amiotrófica/fisiopatologia , Animais , Modelos Animais de Doenças , Mutação , Degeneração Neural/genética , Superóxido Dismutase-1/metabolismo
12.
Commun Biol ; 4(1): 5, 2021 01 04.
Artigo em Inglês | MEDLINE | ID: mdl-33398110

RESUMO

Previous work has revealed that progerin-lamin A binding inhibitor (JH4) can ameliorate pathological features of Hutchinson-Gilford progeria syndrome (HGPS) such as nuclear deformation, growth suppression in patient's cells, and very short life span in an in vivo mouse model. Despite its favorable effects, JH4 is rapidly eliminated in in vivo pharmacokinetic (PK) analysis. Thus, we improved its property through chemical modification and obtained an optimized drug candidate, Progerinin (SLC-D011). This chemical can extend the life span of LmnaG609G/G609G mouse for about 10 weeks and increase its body weight. Progerinin can also extend the life span of LmnaG609G/+ mouse for about 14 weeks via oral administration, whereas treatment with lonafarnib (farnesyl-transferase inhibitor) can only extend the life span of LmnaG609G/+ mouse for about two weeks. In addition, progerinin can induce histological and physiological improvement in LmnaG609G/+ mouse. These results indicate that progerinin is a strong drug candidate for HGPS.


Assuntos
Progéria/tratamento farmacológico , Adolescente , Animais , Criança , Modelos Animais de Doenças , Avaliação Pré-Clínica de Medicamentos , Feminino , Células HEK293 , Humanos , Lamina Tipo A/antagonistas & inibidores , Masculino , Camundongos , Cultura Primária de Células
13.
J Microbiol Biotechnol ; 20(1): 15-20, 2010 Jan.
Artigo em Inglês | MEDLINE | ID: mdl-20134228

RESUMO

A Gram negative, aerobic, nonspore-forming, straight or curved rod-shaped bacterium, designated Gsoil 317T, was isolated from soil of a ginseng field in Pocheon Province (South Korea) and was characterized using a polyphasic approach. Cells were dimorphic, with stalk (or prostheca) and nonmotile or nonstalked and motile, by means of a single polar flagellum. Comparative analysis of 16S rRNA gene sequences revealed that strain Gsoil 317T was most closely related to Caulobacter mirabilis LMG 24261T (97.2%), Caulobacter fusiformis ATCC 15257T (97.1%), Caulobacter segnis LMG 17158T (97.0%), Caulobacter vibrioides DSM 9893T (96.8%), and Caulobacter henricii ATCC 15253T (96.7%). The sequence similarities to any other recognized species within Alphaproteobacteria were less than 96.0%. The detection of Q-10 as the major respiratory quinone and a fatty acid profile with summed feature 7 (C18:1 omega7c and/or C18:1 omega9t and/or C18:1 omega12t; 56.6%) and C16:0 (15.9%) as the major fatty acids supported the affiliation of strain Gsoil 317T to the genus Caulobacter. The G+C content of the genomic DNA was 65.5 mol%. DNA-DNA hybridization experiments showed that the DNA-DNA relatedness values between strain Gsoil 317T and its closest phylogenetic neighbors were below 11%. On the basis of its phenotypic properties and phylogenetic distinctiveness, strain Gsoil 317T should be classified as representing a novel species in the genus Caulobacter, for which the name Caulobacter ginsengisoli sp. nov. is proposed. The type strain is Gsoil 317T (=KCTC 12788T= DSM 18695T).


Assuntos
Caulobacter/classificação , Caulobacter/isolamento & purificação , Panax/microbiologia , Microbiologia do Solo , Composição de Bases , Caulobacter/genética , Caulobacter/metabolismo , DNA Bacteriano/genética , DNA Ribossômico/genética , Ácidos Graxos/metabolismo , Dados de Sequência Molecular , RNA Ribossômico 16S/genética
14.
Cell Death Dis ; 10(2): 107, 2019 02 06.
Artigo em Inglês | MEDLINE | ID: mdl-30728349

RESUMO

p53-mediated cellular senescence has been intensively investigated, because it is important for tumor suppressive function. In addition, p16/INK4A is well known to be critical for cellular senescence. However, detailed molecular mechanism or relevance between p53 and p16-mediated senescence has not been demonstrated yet. Here we show that p53 induces p16 through Lamin A/C stabilization via direct interaction. Stabilized Lamin A/C promotes degradation of BMI-1 and MEL-18 (Polycomb repressor complex 1, PRC1), which sequesters p16 promotor. Increased p53 can reduce BMI-1/MEL-18 and induce p16 expression via Lamin A/C. Elimination of Lamin A/C can abolish p53-induced p16 expression and BMI-1/MEL-18 reduction. As Lamin A/C expression is increased during cell differentiation, this mechanism seems to be very useful for selective induction of senescence in non-stem cells. Our results suggest that Lamin A/C-p53 network is important for p16/INK4A-mediated cellular senescence.


Assuntos
Lamina Tipo A/metabolismo , Laminas/metabolismo , Proteína Supressora de Tumor p53/metabolismo , Linhagem Celular Tumoral , Núcleo Celular/metabolismo , Senescência Celular/fisiologia , Criança , Inibidor p16 de Quinase Dependente de Ciclina/biossíntese , Inibidor p16 de Quinase Dependente de Ciclina/genética , Dano ao DNA , Feminino , Fibroblastos/metabolismo , Técnicas de Silenciamento de Genes , Células HCT116 , Células HEK293 , Humanos , Proteína Quinase 7 Ativada por Mitógeno/metabolismo , Complexo Repressor Polycomb 1/metabolismo , Estabilidade Proteica , Transcrição Gênica , Transfecção , Proteína Supressora de Tumor p53/genética
15.
BMB Rep ; 51(7): 327-337, 2018 Jul.
Artigo em Inglês | MEDLINE | ID: mdl-29764566

RESUMO

Lamin A and its alternative splicing product Lamin C are the key intermediate filaments (IFs) of the inner nuclear membrane intermediate filament. Lamin A/C forms the inner nuclear mesh with Lamin B and works as a frame with a nuclear shape. In addition to supporting the function of nucleus, nuclear lamins perform important roles such as holding the nuclear pore complex and chromatin. However, mutations on the Lamin A or Lamin B related proteins induce various types of human genetic disorders and diseases including premature aging syndromes, muscular dystrophy, lipodystrophy and neuropathy. In this review, we briefly overview the relevance of genetic mutations of Lamin A, human disorders and laminopathies. We also discuss a mouse model for genetic diseases. Finally, we describe the current treatment for laminopathies. [BMB Reports 2018; 51(7): 327-337].


Assuntos
Lamina Tipo A/metabolismo , Lipodistrofia/patologia , Distrofias Musculares/patologia , Animais , Modelos Animais de Doenças , Inibidores Enzimáticos/uso terapêutico , Humanos , Lamina Tipo A/química , Lamina Tipo A/genética , Lipodistrofia/genética , Distrofias Musculares/genética , Mutação , Progéria/tratamento farmacológico , Progéria/genética , Progéria/patologia
16.
J Microbiol Biotechnol ; 28(6): 987-996, 2018 Jun 28.
Artigo em Inglês | MEDLINE | ID: mdl-29642286

RESUMO

Bacterial programmed cell death is regulated by the toxin-antitoxin (TA) system. YhaV (toxin) and Pr1F (antitoxin) have been recently identified as a type II TA system in Escherichia coli. YhaV homologs have conserved active residues within the C-terminus, and to characterize the function of this region, we purified native YhaV protein (without denaturing) and constructed YhaV proteins of varying lengths. Here, we report a new low-temperature method of purifying native YhaV, which is notable given the existing challenges of purifying this highly toxic protein. The secondary structures and thermostability of the purified native protein were characterized and no significant structural destruction was observed, suggesting that the observed inhibition of cell growth in vivo was not the result of structural protein damage. However, it has been reported that excessive levels of protein expression may result in protein misfolding and changes in cell growth and mRNA stability. To exclude this possibility, we used an [³5S]-methionine prokaryotic cell-free protein synthesis system in vitro in the presence of purified YhaV, and two C-terminal truncated forms of this protein (YhaV-L and YhaV-S). Our results suggest that the YhaV C-terminal region is essential for mRNA interferase activity, and the W143 or H154 residues may play an analogous role to Y87 of RelE.


Assuntos
Toxinas Bacterianas/toxicidade , Proteínas de Escherichia coli/toxicidade , Escherichia coli/efeitos dos fármacos , Escherichia coli/crescimento & desenvolvimento , Toxinas Bacterianas/química , Toxinas Bacterianas/genética , Toxinas Bacterianas/isolamento & purificação , Análise Mutacional de DNA , Escherichia coli/genética , Proteínas de Escherichia coli/química , Proteínas de Escherichia coli/genética , Proteínas de Escherichia coli/isolamento & purificação , Expressão Gênica , Estabilidade Proteica , Estrutura Secundária de Proteína , Estabilidade de RNA , Proteínas Recombinantes/química , Proteínas Recombinantes/genética , Proteínas Recombinantes/isolamento & purificação , Proteínas Recombinantes/toxicidade
17.
Mol Cancer Res ; 16(6): 935-946, 2018 06.
Artigo em Inglês | MEDLINE | ID: mdl-29545477

RESUMO

Quinacrine (QNC), antiprotozoan drug commonly used against Malaria and Giardiasis, has been recently tried for rheumatics and prion diseases via drug repositioning. In addition, several reports suggest antitumor effects of QNC through suppression of NF-κB and activation of p53. This study demonstrates the anticancer effect of QNC via a novel pathway through the elimination of checkpoint kinase 1/2 (Chk1/2) under p53-inactivated conditions. Inhibition of p53 by PFT-α or siRNA promotes QNC-induced apoptosis in normal fibroblast and p53-intact cancer cells. Considering that Chk1/2 kinases exert an essential role in the control of cell cycle, inhibition of Chk1/2 by QNC may induce cell death via uncontrolled cell cycle progression. Indeed, QNC reduces Chk1/2 expression under p53-impaired cancer cells and induces cell death in the G2-M phase. QNC increases the binding between p-Chk1/2 and ß-TrCP and promotes proteasome-dependent degradation. Moreover, QNC treatment displayed antitumor effects in a Villin-Cre;p53+/LSL-R172H intestinal cancer mouse model system as well as HCT116 p53-/- xenografts.Implications: QNC has been used for the past over 70 years without obvious side effects, as such it is a plausible drug candidate for relapsed cancers, small-cell lung cancer, breast cancer as well as various p53-inactivated human malignancies. Mol Cancer Res; 16(6); 935-46. ©2018 AACR.


Assuntos
Antimaláricos/uso terapêutico , Quinase 1 do Ponto de Checagem/genética , Neoplasias/tratamento farmacológico , Quinacrina/uso terapêutico , RNA Interferente Pequeno/metabolismo , Proteína Supressora de Tumor p53/genética , Animais , Antimaláricos/farmacologia , Humanos , Camundongos , Camundongos Nus , Neoplasias/genética , Neoplasias/patologia , Quinacrina/farmacologia , Transfecção
18.
Mol Cancer Ther ; 17(11): 2271-2284, 2018 11.
Artigo em Inglês | MEDLINE | ID: mdl-30135214

RESUMO

Neurofibromatosis type 2 (NF2) syndrome is a very rare human genetic disease, and there has been no proper treatment for it until now. In our recent study, it has been reported that the loss of NF2 activates MAPK signaling through reduction of RKIP in a mesothelioma model. Here, we show that loss of NF2 induces reduction of the TGFß receptor 2 (TßR2) expression, and an overwhelming expression of TGFß receptor 1 (TßR1) is activated by physical stimuli such as pressure or heavy materials. Activated TßR1 induces the phosphorylation and degradation of RKIP. RKIP reduction consequently results in MAPK activation as well as Snail-mediated p53 suppression and occurrence of EMT in NF2-deficient cells by physical stimuli. Thus, TßR1 kinase inhibitors restore cell differentiation and induce growth suppression in NF2-deficient Schwannoma cell line and MEF. Moreover, TEW7197, a specific TßR1 kinase inhibitor, reduces tumor formation in the NF2-model mouse (Postn-Cre;NF2f/f). Gene expression profiling reveals that TEW7197 treatment induces the expression of lipid metabolism-related gene set, such as NF2-restored cells in HEI-193 (NF2-deficient Schwannoma). Our results indicate that reduction or deletion of TßR2 or NF2 induces the TßR1-mediated oncogenic pathway, and therefore inhibition of the unbalanced TGFß signaling is a putative strategy for NF2-related cancers (NF2 syndrome and mesothelioma) and TßR2-mutated advanced cancers. Mol Cancer Ther; 17(11); 2271-84. ©2018 AACR.


Assuntos
Neurofibromatose 2/tratamento farmacológico , Neurofibromina 2/deficiência , Oncogenes , Receptor do Fator de Crescimento Transformador beta Tipo I/antagonistas & inibidores , Receptor do Fator de Crescimento Transformador beta Tipo I/metabolismo , Transdução de Sinais , Fator de Crescimento Transformador beta/metabolismo , Animais , Carcinogênese , Diferenciação Celular/efeitos dos fármacos , Linhagem Celular Tumoral , Proliferação de Células/efeitos dos fármacos , Modelos Animais de Doenças , Transição Epitelial-Mesenquimal , Humanos , Camundongos , Neurilemoma/patologia , Neurofibromina 2/metabolismo , Proteína de Ligação a Fosfatidiletanolamina/metabolismo , Inibidores de Proteínas Quinases/farmacologia , Inibidores de Proteínas Quinases/uso terapêutico , Receptor do Fator de Crescimento Transformador beta Tipo II/metabolismo , Dióxido de Silício , Suínos
19.
J Microbiol Biotechnol ; 17(8): 1291-9, 2007 Aug.
Artigo em Inglês | MEDLINE | ID: mdl-18051597

RESUMO

Two beta-1,4-glucanases (DI and DIII fractions) were purified to homogeneity from the culture filtrate of a cellulolytic bacteria, Cellulomonas sp. CS1-1, which was classified as a novel species belonging to Cellulomonas uda based on chemotaxanomic and phylogenetic analyses. The molecular mass was estimated as 50,000 Da and 52,000 Da for DI and DIII, respectively. Moreover, DIII was identified as a glycoprotein with a pI of 3.8, and DI was identified as a non-glycoprotein with a pI of 5.3. When comparing the ratio of the CMC-saccharifying activity and CMC-liquefying activity, DI exhibited a steep slope, characteristic of an endoglucanase, whereas DIII exhibited a low slope, characteristic of an exoglucanase. The substrate specificity of the purified enzymes revealed that DI efficiently hydrolyzed CMC as well as xylan, whereas DIII exhibited a high activity on microcrystalline celluloses, such as Sigmacells. A comparison of the hydrolysis patterns for pNP-glucosides (DP 2-5) using an HPLC analysis demonstrated that the halosidic bond 3 from the nonreducing end was the preferential cleavage site for DI, whereas bond 2, from which the cellobiose unit is split off, was the preferential cleavage site for DIII. The partial N-terminal amino acid sequences for the purified enzymes were 1Ala-Gly-Ser-Thr-Leu-Gln-Ala-Ala-Ala-Ser-Glu-Ser-Gly-Arg-Tyr15- for DI and 1Ala-Asp-Ser-Asp-Phe-Asn-Leu-Tyr-Val-Ala-Glu-Asn-Ala-Met-Lys15- for DIII. The apparent sequences exhibited high sequence similarities with other bacterial beta-1,4-glucanases as well as beta-1,4-xylanases.


Assuntos
Celulase/isolamento & purificação , Celulase/metabolismo , Cellulomonas/enzimologia , Glucana 1,4-beta-Glucosidase/isolamento & purificação , Glucana 1,4-beta-Glucosidase/metabolismo , Sequência de Aminoácidos , Proteínas de Bactérias/química , Proteínas de Bactérias/isolamento & purificação , Proteínas de Bactérias/metabolismo , Carboximetilcelulose Sódica/metabolismo , Celulase/química , Cellulomonas/classificação , Cellulomonas/genética , Cellulomonas/isolamento & purificação , Celulose/metabolismo , DNA Bacteriano/química , DNA Bacteriano/genética , DNA Ribossômico/química , DNA Ribossômico/genética , Glucana 1,4-beta-Glucosidase/química , Ponto Isoelétrico , Dados de Sequência Molecular , Peso Molecular , Filogenia , RNA Ribossômico 16S/genética , Análise de Sequência de DNA , Análise de Sequência de Proteína , Homologia de Sequência de Aminoácidos , Especificidade por Substrato , Xilanos/metabolismo
20.
J Microbiol Biotechnol ; 17(6): 913-8, 2007 Jun.
Artigo em Inglês | MEDLINE | ID: mdl-18050908

RESUMO

A Gram-positive, aerobic, rod-shaped, nonmotile, endospore-forming bacterium, designated Gsoil 349T, was isolated from soil of a ginseng field and characterized using a polyphasic approach. Comparative analysis of 16S rRNA gene sequences revealed that the strain Gsoil 349T belongs to the family Paenibacillaceae, and the sequence showed closest similarity with Cohnella thermotolerans DSM 17683T (94.1%) and Cohnella hongkongensis DSM 17642T (93.6%). The strain showed less than 91.3% 16S rRNA gene sequence similarity with Paenibacillus species. In addition, the presence of MK-7 as the major menaquinone and anteiso-C(15:0), iso-C(16:0), and C(16:0) as major fatty acids suggested its affiliation to the genus Cohnella. The G+C content of the genomic DNA was 53.4 mol%. On the basis of its phenotypic characteristics and phylogenetic distinctiveness, strain Gsoil 349T should be treated as a novel species within the genus Cohnella for which the name Cohnella panacarvi sp. nov. is proposed. The type strain is Gsoil 349T (=KCTC 13060T = DSM 18696T).


Assuntos
Bactérias Gram-Positivas Formadoras de Endosporo/classificação , Panax/microbiologia , Microbiologia do Solo , Xilanos/metabolismo , Composição de Bases , Ácidos Graxos/análise , Bactérias Gram-Positivas Formadoras de Endosporo/isolamento & purificação , Bactérias Gram-Positivas Formadoras de Endosporo/metabolismo , Panax/crescimento & desenvolvimento , Fenótipo , Filogenia , RNA Ribossômico 16S/genética , Vitamina K 2/análise
SELEÇÃO DE REFERÊNCIAS
DETALHE DA PESQUISA