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1.
Cancer Sci ; 115(9): 2972-2984, 2024 Sep.
Artículo en Inglés | MEDLINE | ID: mdl-38951133

RESUMEN

Serum laminin-γ2 monomer (Lm-γ2m) is a potent predictive biomarker for hepatocellular carcinoma (HCC) onset in patients with hepatitis C infection who achieve a sustained virologic response with liver cirrhosis (LC) and for the onset of extrahepatic metastases in early-stage HCC. Although Lm-γ2m involvement in late-stage cancer progression has been well investigated, its precise roles in HCC onset remain to be systematically investigated. Therefore, we analyzed an HCC model, human hepatocytes and cholangiocytes, and surgically resected liver tissues from patients with HCC to understand the roles of Lm-γ2m in HCC onset. Ck-19- and EpCAM-positive hepatic progenitor cells (HPCs) in the liver of pdgf-c transgenic HCC mouse model with ductular reaction showed ectopic expression of Lm-γ2m. Forced expression of Lm-γ2m in hepatocytes adjacent to HPCs resulted in enhanced tumorigenicity, cell proliferation, and migration in immortalized hepatocytes, but not in cholangiocytes in vitro. Further, pharmacological inhibition of epidermal growth factor receptor (EGFR) and c-Jun activator JNK suppressed Lm-γ2m-induced hepatocyte transformation, suggesting the involvement of EGFR/c-Jun signaling in the transformation, leading to HCC development. Finally, immunohistochemical staining of HCC tissues revealed a high level of Lm-γ2 expression in the HPCs of the liver with ductular reaction in normal liver adjacent to HCC tissues. Overall, HPC-derived Lm-γ2m in normal liver with ductular reaction acts as a paracrine growth factor on surrounding hepatocytes and promotes their cellular transformation through the EGFR/c-Jun signaling pathway. Furthermore, this is the first report on Lm-γ2m expression detected in the normal liver with ductular reaction, a human precancerous lesion of HCC.


Asunto(s)
Carcinoma Hepatocelular , Transformación Celular Neoplásica , Receptores ErbB , Hepatocitos , Laminina , Neoplasias Hepáticas , Humanos , Hepatocitos/metabolismo , Animales , Ratones , Neoplasias Hepáticas/metabolismo , Neoplasias Hepáticas/patología , Carcinoma Hepatocelular/metabolismo , Carcinoma Hepatocelular/patología , Transformación Celular Neoplásica/metabolismo , Laminina/metabolismo , Receptores ErbB/metabolismo , Ratones Transgénicos , Transducción de Señal , Proliferación Celular , Masculino , Movimiento Celular , Hígado/metabolismo , Hígado/patología
2.
Environ Microbiome ; 18(1): 41, 2023 May 10.
Artículo en Inglés | MEDLINE | ID: mdl-37165459

RESUMEN

Understanding the variability of microbial niches and their interaction with abiotic and biotic factors in the Arctic can provide valuable insights into microbial adaptations to extreme environments. This study investigates the structure and diversity of soil bacterial communities obtained from sites with varying vegetation coverage and soil biogeochemical properties in the low Arctic tundra and explores how bacteria interact under different environmental parameters. Our findings reveal differences in bacterial composition and abundance among three bacterial niche breadths (specialists, common taxa, and generalists). Co-occurrence network analysis revealed Rhizobiales and Ktedonobacterales as keystone taxa that connect and support other microbes in the habitat. Low-elevation indicators, such as vascular plants and moisture content, were correlated with two out of three generalist modular hubs and were linked to a large proportion of generalists' distribution (18%). Structural equation modeling revealed that generalists' distribution, which influenced the remaining microbial communities, was mainly regulated by vegetation coverage as well as other abiotic and biotic factors. These results suggest that elevation-dependent environmental factors directly influence microbial community structure and module formation through the regulation of generalists' distribution. Furthermore, the distribution of generalists was mainly affected by macroenvironment filtering, whereas the distribution of specialists was mainly affected by microenvironment filtering (species-engineered microbial niche construction). In summary, our findings highlight the strong top-down control exerted by vegetation on generalists' distribution, which in turn shapes the overall microbial community structure in the low Arctic tundra.

3.
FEMS Microbiol Ecol ; 99(2)2023 01 24.
Artículo en Inglés | MEDLINE | ID: mdl-36581318

RESUMEN

Beta-diversity partitioning has shown that the nestedness component is developed with environmental stress in a variety of taxa. However, soil fungal community may maintain its turnover components in contrast to the development of plants' nestedness component, and the potential causes remain unclear. To investigate the process of species turnover of soil fungi along a stress gradient in the Arctic, we divided species turnover component into sub-components: ßsim_hete and ßsim_homo representing species turnover with and without a change in the guilds, respectively. The results indicate that fungal communities maintain their turnover components, unlike plant communities; however, their ßsim_hete increased under stressful conditions. Additionally, GDM analysis showed that ßsim_hete was mainly explained by stress gradient and plant nestedness, suggesting that the functionality of soil fungi was ecologically filtered by environmental stress and plant community structure. The discordant trend of beta-diversity values between plant and fungi (i.e. development of plant nestedness and maintenance of fungal turnover) is possibly not caused by different assembly rules working in parallel on the two taxa, but according to an ecological rule that reflects plant-fungal interaction.


Asunto(s)
Biodiversidad , Plantas , Hongos/genética , Suelo/química , Microbiología del Suelo
4.
Cancer Sci ; 114(2): 348-356, 2023 Feb.
Artículo en Inglés | MEDLINE | ID: mdl-36336966

RESUMEN

Strategies to develop cancer therapies using inhibitors that target matrix metalloproteinases (MMPs), particularly membrane type-1 MMP (MT1-MMP), have failed. This is predominantly attributed to the specificity of MMP inhibitors and numerous functions of MMPs; therefore, targeting substrates with such broad specificity can lead to off-target effects. Thus, new drug development for cancer therapeutics should focus on the ability of MT1-MMP to break down substrates, such as functional cell membrane proteins, to regulate the functions of these proteins that promote tumor malignancy. In this review, we discuss the mechanism by which proteolysis of cell surface proteins by MT1-MMP promotes progression of malignant tumor cells. In addition, we discuss the two protein fragments generated by limited cleavage of erythropoietin-producing hepatoma receptor tyrosine kinase A2 (EphA2-NF, -CF), which represent a promising basis for developing new cancer therapies and diagnostic techniques.


Asunto(s)
Proteínas de la Membrana , Neoplasias , Humanos , Proteolisis , Proteínas de la Membrana/metabolismo , Metaloproteinasa 14 de la Matriz/metabolismo , Metaloendopeptidasas/metabolismo
5.
Sci Rep ; 12(1): 2450, 2022 02 14.
Artículo en Inglés | MEDLINE | ID: mdl-35165334

RESUMEN

The neuroplastic mechanism of sex reversal in the fish brain remains unclear due to the difficulty in identifying the key neurons involved. Mozambique tilapia show different reproductive behaviours between sexes; males build circular breeding nests while females hold and brood fertilized eggs in their mouth. In tilapia, gonadotropin-releasing hormone 3 (GnRH3) neurons, located in the terminal nerve, regulate male reproductive behaviour. Mature males have more GnRH3 neurons than mature females, and these neurons have been indicated to play a key role in the androgen-induced female-to-male sex reversal of the brain. We aimed to elucidate the signalling pathway involved in the androgen-induced increase in GnRH3 neurons in mature female tilapia. Applying inhibitors to organotypic cultures of brain slices, we showed that the insulin-like growth factor (IGF)-1 receptor (IGF-1R)/PI3K/AKT/mTOR pathway contributed to the androgen-induced increase in GnRH3 neurons. The involvement of IGF-1 and IGF-1R in 11-ketotestosterone (11-KT)-induced development of GnRH3 neurons was supported by an increase in Igf-1 mRNA shortly after 11-KT treatment, the increase of GnRH3 neurons after IGF-1 treatment and the expression of IGF-1R in GnRH3 neurons. Our findings highlight the involvement of IGF-1 and its downstream signalling pathway in the sex reversal of the tilapia brain.


Asunto(s)
Encéfalo/metabolismo , Hormona Liberadora de Gonadotropina/metabolismo , Metiltestosterona/farmacología , Neuronas/metabolismo , Fosfatidilinositol 3-Quinasas/metabolismo , Ácido Pirrolidona Carboxílico/análogos & derivados , Receptor IGF Tipo 1/metabolismo , Reproducción/efectos de los fármacos , Transducción de Señal/efectos de los fármacos , Serina-Treonina Quinasas TOR/metabolismo , Animales , Encéfalo/efectos de los fármacos , Femenino , Factor I del Crecimiento Similar a la Insulina/farmacología , Masculino , Neuronas/efectos de los fármacos , Proteínas Proto-Oncogénicas c-akt/metabolismo , Ácido Pirrolidona Carboxílico/metabolismo , Testosterona/análogos & derivados , Testosterona/farmacología , Tilapia
6.
FEMS Microbiol Ecol ; 97(10)2021 10 01.
Artículo en Inglés | MEDLINE | ID: mdl-34494103

RESUMEN

Metabarcoding technologies for soil fungal DNA pools have enabled to capture the diversity of fungal community and the agreement of their ß-diversity with plant ß-diversity. However, processes underlying the synchrony of the aboveground-belowground biodiversity is still unclear. By using partitioning methods for plant ß-diversity, this study explored the process driving synchrony in tundra ecosystems, in which drastic vegetation shifts are observed with climate warming. Our methods based on Baselga's partitioning enabled the division of plant ß-diversity into two phenomena and three functional components. Correlation of fungal ß-diversity with the components of plant ß-diversity showed that the spatial replacement of fungi was promoted by plant species turnover, in particular, plant species turnover with functional exchange. In addition, spatial variety of graminoid or forbs species, rather than shrubs, enhanced fungal ß-diversity. These results suggest the importance of small-scale factors such as plant-fungal interactions or local environments modified by plants for the fungal community assemblage. The process-based understanding of community dynamics of plants and fungi allows us to predict the ongoing shrub encroachment in the Arctic region, which could weaken the aboveground-belowground synchrony.


Asunto(s)
Ecosistema , Suelo , Regiones Árticas , Biodiversidad , Hongos/genética , Tundra
7.
J Chem Phys ; 155(2): 024302, 2021 Jul 14.
Artículo en Inglés | MEDLINE | ID: mdl-34266257

RESUMEN

Icosahedral noble-metal 13-atom nanoclusters (NCs) can form connected structures, which can be regarded as superatomic molecules, by vertex sharing. However, there have been very few reports on the superatomic molecules formed using silver (Ag) as the base element. In this study, we synthesized [Ag23Pd2(PPh3)10Cl7]0 (Pd = palladium, PPh3 = triphenylphosphine, Cl = chloride), in which two icosahedral 13-atom NCs are connected, and elucidated its geometric and electronic structures to clarify what type of superatomic molecules can be synthesized. The results revealed that [Ag23Pd2(PPh3)10Cl7]0 is a synthesizable superatomic molecule. Single crystal x-ray diffraction analysis showed that the metal-metal distances in and between the icosahedral structures of [Ag23Pd2(PPh3)10Cl7]0 are slightly shorter than those of previously reported [Ag23Pt2(PPh3)10Cl7]0, whereas the metal-PPh3 distances are slightly longer. On the basis of several experiments and density functional theory calculations, we concluded that [Ag23Pd2(PPh3)10Cl7]0 and previously reported [Ag23Pt2(PPh3)10Cl7]0 are more stable than [Ag25(PPh3)10Cl7]2+ because of their stronger superatomic frameworks (metal cores). These findings are expected to lead to clear design guidelines for creation of new superatomic molecules.

8.
Chemistry ; 26(69): 16149, 2020 Dec 09.
Artículo en Inglés | MEDLINE | ID: mdl-33119138

RESUMEN

Invited for the cover of this issue is the group of Yuichi Negishi at Tokyo University of Science. The image depicts the alloy nanoclusters reported in this review. Read the full text of the article at 10.1002/chem.202001877.

9.
Front Microbiol ; 11: 1372, 2020.
Artículo en Inglés | MEDLINE | ID: mdl-32754122

RESUMEN

Dimethyl sulfide (DMS) is an important component of the global sulfur cycle as it is the most abundant sulfur compound that is emitted via the ocean surface to the atmosphere. Dimethylsulfoniopropionate (DMSP), the precursor of DMS, is mainly produced by phytoplankton and is degraded by marine bacteria. To reveal the role of bacteria in the regulation of DMSP degradation and DMS production, mesocosm and field studies were performed in the Sanriku Coast on the Pacific Ocean in northeast Japan. The responsible bacteria for the transformation of DMSP to DMS and the assimilation of DMSP were monitored, and the genes encoding DMSP lyase (dddD and dddP) and DMSP demethylase (dmdA) were analyzed. The mesocosm study showed that the dmdA subclade D was the dominant DMSP degradation gene in the free-living (FL) and particle-associated (PA) fractions. The dddD gene was found in higher abundance than the dddP gene in all the tested samples. Most importantly, DMS concentration was positively correlated with the abundance of the dddD gene. These results indicated that bacteria possessing dmdA and dddD genes were the major contributors to the DMSP degradation and DMS production, respectively. The genes dmdA subclade D and dddP were abundant in the Tsugaru Warm (TW) Current, while the dmdA subclade C/2 and dddD genes were dominant in the Oyashio (OY) Current. Functional gene network analysis also showed that the DMSP degradation genes were divided into OY and TW Current-related modules, and genes sharing similar functions were clustered in the same module. Our data suggest that environmental fluctuations resulted in habitat filtering and niche partitioning of bacteria possessing DMSP degradation genes. Overall, our findings provide novel insights into the distribution and abundance of DMSP degradation genes in a coastal region with different water current systems.

10.
Chemistry ; 26(69): 16150-16193, 2020 Dec 09.
Artículo en Inglés | MEDLINE | ID: mdl-32453462

RESUMEN

Metal nanoclusters (NCs) have a particle size of about one nanometer, which makes them the smallest unit that can give a function to a substance. In addition, metal NCs possess physical and chemical properties that are different from those of the corresponding bulk metals. Metal NCs with such characteristics are expected to be important for use in nanotechnology. Research on the precise synthesis of metal NCs and elucidation of their physical/chemical properties and functions is being actively conducted. When metal NCs are alloyed, it is possible to obtain further various electronic and geometrical structures and functions. Thus, research on alloy NCs has become a hot topic in the study of metal NCs and the number of publications on alloy NCs has increased explosively in recent years. Such publications have provided much insight into the effects of alloying on the electronic structure and function of metal NCs. However, the rapid increase in knowledge has made it difficult for researchers (especially those new to the field) to grasp all of it. Therefore, in this review, we summarize the reported chemical composition, geometrical structure, electronic structure, and physical and chemical properties of Aun-x Mx (SR)m , Agn-x Mx (SR)m , Aun-x Mx (PR3 )l (SR)m , and Agn-x Mx (PR3 )l (SR)m (Au=gold, Ag=silver, M=heteroatom, PR3 =phosphine, and SR=thiolate) NCs. This review is expected to help researchers understand the characteristics of alloy NCs and lead to clear design guidelines to develop new alloy NCs with intended functions.

11.
Nanoscale Adv ; 2(2): 669-678, 2020 Feb 18.
Artículo en Inglés | MEDLINE | ID: mdl-36133224

RESUMEN

Although Pt is extensively used as a catalyst to purify automotive exhaust gas, it is desirable to reduce Pt consumption through size reduction because Pt is a rare element and an expensive noble metal. In this study, we successfully loaded a Pt17 cluster on γ-alumina (γ-Al2O3) (Pt17/γ-Al2O3) using [Pt17(CO)12(PPh3)8]Cl n (n = 1, 2) as a precursor. In addition, we demonstrated that Pt is not present in the form of an oxide in Pt17/γ-Al2O3 but instead has a framework structure as a metal cluster. Moreover, we revealed that Pt17/γ-Al2O3 exhibits higher catalytic activity for carbon monoxide and propylene oxidation than γ-Al2O3-supported larger Pt nanoparticles (PtNP/γ-Al2O3) prepared using the conventional impregnation method. Recently, our group discovered a simple method for synthesizing the precursor [Pt17(CO)12(PPh3)8]Cl n . Furthermore, Pt17 is a Pt cluster within the size range associated with high catalytic activity. By combining our established synthesis and loading methods, other groups can conduct further research on Pt17/γ-Al2O3 to explore its catalytic activities in greater depth.

12.
Front Microbiol ; 10: 2722, 2019.
Artículo en Inglés | MEDLINE | ID: mdl-31849876

RESUMEN

Greater diversity of eukaryotic phytoplankton than expected has been revealed recently through molecular techniques, but little is known about their temporal dynamics or fate in the open ocean. Here, we examined size-fractionated eukaryotic phytoplankton communities from the surface to abyssopelagic zone (5,000 m) throughout the year, by tracking sequence variants of the 18S rRNA gene in the western subtropical North Pacific. The oceanographic conditions were divided into two periods, stratification and mixing, between which the surface phytoplankton community differed. During the mixing period, the abundance of large phytoplankton (≥3 µm) increased, with diatoms and putative Pseudoscourfieldia marina dominating this fraction. Picophytoplankton (<3 µm) also increased during the mixing period and were dominated by Mamiellophyceae. Taxa belonging to prasinophytes (including Ps. marina and Mamiellophyceae) were observed in the epipelagic zone throughout the year, and thus likely seeded the seasonal bloom that occurred during the mixing period. In contrast, diatoms observed during the mixing period mostly represented taxa unique to that period, including coastal species. Numerical particle backtracking experiments indicated that water masses in the surface layer could be transported from coastal areas to the study site. Gene sequences of coastal diatoms were present in the abyssopelagic zone. Therefore, allochthonous species drove the seasonal bloom and could be transported to deep waters. In the abyssopelagic zone, the relative abundance of Ps. marina in deep waters was similar to or higher than that of diatoms during the mixing period. Among picophytoplankton, Mamiellophyceae made up a significant fraction in the abyssopelagic zone, suggesting that prasinophytes are also involved in carbon export. Our molecular survey showed that these previously overlooked phytoplankton species could contribute significantly to the seasonal bloom and biological pump in the subtropical open ocean.

13.
J Microbiol ; 57(5): 413-422, 2019 May.
Artículo en Inglés | MEDLINE | ID: mdl-30806980

RESUMEN

Although culture-independent studies have shown the presence of Verrucomicrobia in the deep sea, verrucomicrobial strains from deep-sea environments have been rarely cultured and characterized. Recently, Rubritalea profundi SAORIC-165T, a psychrophilic bacterium of the phylum Verrucomicrobia, was isolated from a depth of 2,000 m in the northwestern Pacific Ocean. In this study, the genome sequence of R. profundi SAORIC-165T, the first deep-sea verrucomicrobial isolate, is reported with description of the genome properties and comparison to surface-borne Rubritalea genomes. The draft genome consisted of four contigs with an entire size of 4,167,407 bp and G+C content of 47.5%. The SAORIC-165T genome was predicted to have 3,844 proteincoding genes and 45 non-coding RNA genes. The genome contained a repertoire of metabolic pathways, including the Embden-Meyerhof-Parnas pathway, pentose phosphate pathway, tricarboxylic acid cycle, assimilatory sulfate reduction, and biosynthesis of nicotinate/nicotinamide, pantothenate/coenzyme A, folate, and lycopene. The comparative genomic analyses with two surface-derived Rubritalea genomes showed that the SAORIC-165T genome was enriched in genes involved in transposition of mobile elements, signal transduction, and carbohydrate metabolism, some of which might be related to bacterial enhancement of ecological fitness in the deep-sea environment. Amplicon sequencing of 16S rRNA genes from the water column revealed that R. profundi-related phylotypes were relatively abundant at 2,000 m and preferred a particle-associated life style in the deep sea. These findings suggest that R. profundi represents a genetically unique and ecologically relevant verrucomicrobial group well adapted to the deep-sea environment.


Asunto(s)
Sedimentos Geológicos/microbiología , Agua de Mar/microbiología , Verrucomicrobia/clasificación , Verrucomicrobia/genética , Técnicas de Tipificación Bacteriana , Composición de Base/genética , Secuencia de Bases , ADN Bacteriano/genética , Genoma Bacteriano/genética , Océano Pacífico , ARN Ribosómico 16S/genética , Verrucomicrobia/aislamiento & purificación
14.
PLoS One ; 13(8): e0202636, 2018.
Artículo en Inglés | MEDLINE | ID: mdl-30125317

RESUMEN

Planktonic archaea are thought to play an important role in ammonia oxidation in marine environments. Data on the distribution, abundance, and diversity of ammonia oxidizers in the coastal sea-surface microlayer (SML) are lacking, despite previous reports of high abundance of Thaumarchaeota in the SML of estuaries and freshwater lakes. Here, we failed to detect the presence of ammonia-oxidizing bacteria in any of our samples taken from a semi-enclosed marine inlet in Japan. Therefore, we shifted our focus to examine the archaeal community composition as well as the Thaumarchaeota marine group I (MG-I) and ammonia monooxygenase subunit A (amoA) gene copy numbers and composition in the SML and corresponding underlying water (UW, 20 cm). amoA gene copy numbers obtained by quantitative PCR were consistent with the typical values observed in the surface waters of oceanic and coastal environments where nitrification activity has been detected, but the copy numbers were two- to three-fold less than those reported from the surface layers and UW of high mountain lakes. Both amoA and MG-I 16S rRNA gene copy numbers were significantly negatively correlated with chlorophyll-a and transparent exopolymer particle concentrations in the SML. Communities of archaea and ammonia-oxidizing archaea in SML samples collected during low wind conditions (≤5 m s-1) differed the most from those in UW samples, whereas the communities in SML samples collected during high wind conditions were similar to the UW communities. In the SML, low ratios of amoA to MG-I 16S rRNA genes were observed, implying that most of the SML Thaumarchaeota lacked amoA. To our knowledge, our results provide the first comparison of ammonia-oxidizing communities in the coastal SML with those in the UW.


Asunto(s)
Amoníaco/metabolismo , Archaea/genética , Biodiversidad , Oxidorreductasas/genética , Archaea/metabolismo , Bahías/microbiología , Dosificación de Gen/genética , Sedimentos Geológicos/microbiología , Lagos/microbiología , Nitrificación , Oxidación-Reducción , Oxidorreductasas/metabolismo , ARN Ribosómico 16S/genética , Agua de Mar/microbiología
15.
ISME J ; 12(9): 2238-2251, 2018 09.
Artículo en Inglés | MEDLINE | ID: mdl-29884827

RESUMEN

Coral reef ecosystems are highly sensitive to microbial activities that result from dissolved organic matter (DOM) enrichment of their surrounding seawater. However, the response to particulate organic matter (POM) enrichment is less studied. In a microcosm experiment, we tested the response of bacterioplankton to a pulse of POM from the mass-spawning of Orbicella franksi coral off the Caribbean coast of Panama. Particulate organic carbon (POC), a proxy measurement for POM, increased by 40-fold in seawater samples collected during spawning; 68% degraded within 66 h. The elevation of multiple hydrolases presumably solubilized the spawn-derived POM into DOM. A carbon budget constructed for the 275 µM of degraded POC showed negligible change to the concentration of dissolved organic carbon (DOC), indicating that the DOM was readily utilized. Fourier transform ion cyclotron resonance mass spectrometry shows that the DOM pool became enriched with heteroatom-containing molecules, a trend that suggests microbial alteration of organic matter. Our sensitivity analysis demonstrates that bacterial carbon demand could have accounted for a large proportion of the POC degradation. Further, using bromodeoxyuridine immunocapture in combination with 454 pyrosequencing of the 16S ribosomal RNA gene, we surmise that actively growing bacterial groups were the primary degraders. We conclude that coral gametes are highly labile to bacteria and that such large capacity for bacterial degradation and alteration of organic matter has implications for coral reef health and coastal marine biogeochemistry.


Asunto(s)
Antozoos/fisiología , Bacterias/metabolismo , Plancton/metabolismo , Agua de Mar/química , Animales , Bacterias/crecimiento & desarrollo , Bacterias/aislamiento & purificación , Carbono/análisis , Arrecifes de Coral , Material Particulado/análisis , Plancton/crecimiento & desarrollo , Plancton/aislamiento & purificación
16.
Genome Announc ; 4(6)2016 Dec 08.
Artículo en Inglés | MEDLINE | ID: mdl-27932653

RESUMEN

Here, we report the draft genome sequences of Fabibacter sp. strain 4D4 and F. misakiensis strain SK-8T, isolated from surface seawater of a semienclosed inlet.

17.
Gene ; 576(2 Pt 1): 650-6, 2016 Feb 01.
Artículo en Inglés | MEDLINE | ID: mdl-26474747

RESUMEN

The "rare biosphere" consisting of thousands of low-abundance microbial taxa is important as a seed bank or a gene pool to maintain microbial functional redundancy and robustness of the ecosystem. Here we investigated contemporaneous growth of diverse microbial taxa including rare taxa and determined their variability in environmentally distinctive locations along a north-south transect in the Pacific Ocean in order to assess which taxa were actively growing and how environmental factors influenced bacterial community structures. A bromodeoxyuridine-labeling technique in combination with PCR amplicon pyrosequencing of 16S rRNA genes gave 215-793 OTUs from 1200 to 3500 unique sequences in the total communities and 175-299 OTUs nearly 860 to 1800 sequences in the active communities. Unexpectedly, many of the active OTUs were not detected in the total fractions. Among these active but rare OTUs, some taxa (2-4% of rare OTUs) showed much higher abundance (>0.10% of total reads) in the active fraction than in the total fraction, suggesting that their contribution to bacterial community productivity or growth was much larger than that expected from their standing stocks at each location. An ordination plot by the principal component analysis presented that bacterial community compositions among 4 sampling locations and between total and active fractions were distinctive with each other. A redundancy analysis revealed that the variability of community compositions significantly correlated to seawater temperature and dissolved oxygen concentration. Also, a variation partitioning analysis showed that the environmental factors explained 49% of the variability of community compositions and the distance only explained 4.0% of its variability. These results implied very dynamic change of community structures due to environmental filtering. The active bacterial populations are more diverse and spread further in rare biosphere than we have ever seen. This study implied that rare microbes are important as an active part of microbial communities functioning ecosystems.


Asunto(s)
Bromodesoxiuridina/metabolismo , ARN Ribosómico 16S/genética , Microbiología del Agua , Océano Pacífico , Reacción en Cadena de la Polimerasa , Salinidad , Temperatura
18.
Appl Environ Microbiol ; 81(12): 4184-94, 2015 Jun 15.
Artículo en Inglés | MEDLINE | ID: mdl-25862229

RESUMEN

Dimethylsulfoniopropionate (DMSP) is mainly produced by marine phytoplankton but is released into the microbial food web and degraded by marine bacteria to dimethyl sulfide (DMS) and other products. To reveal the abundance and distribution of bacterial DMSP degradation genes and the corresponding bacterial communities in relation to DMS and DMSP concentrations in seawater, we collected surface seawater samples from DMS hot spot sites during a cruise across the Pacific Ocean. We analyzed the genes encoding DMSP lyase (dddP) and DMSP demethylase (dmdA), which are responsible for the transformation of DMSP to DMS and DMSP assimilation, respectively. The averaged abundance (±standard deviation) of these DMSP degradation genes relative to that of the 16S rRNA genes was 33% ± 12%. The abundances of these genes showed large spatial variations. dddP genes showed more variation in abundances than dmdA genes. Multidimensional analysis based on the abundances of DMSP degradation genes and environmental factors revealed that the distribution pattern of these genes was influenced by chlorophyll a concentrations and temperatures. dddP genes, dmdA subclade C/2 genes, and dmdA subclade D genes exhibited significant correlations with the marine Roseobacter clade, SAR11 subgroup Ib, and SAR11 subgroup Ia, respectively. SAR11 subgroups Ia and Ib, which possessed dmdA genes, were suggested to be the main potential DMSP consumers. The Roseobacter clade members possessing dddP genes in oligotrophic subtropical regions were possible DMS producers. These results suggest that DMSP degradation genes are abundant and widely distributed in the surface seawater and that the marine bacteria possessing these genes influence the degradation of DMSP and regulate the emissions of DMS in subtropical gyres of the Pacific Ocean.


Asunto(s)
Bacterias/clasificación , Bacterias/metabolismo , Genes Bacterianos , Consorcios Microbianos , Agua de Mar/microbiología , Compuestos de Sulfonio/metabolismo , Bacterias/aislamiento & purificación , Liasas de Carbono-Azufre/genética , Clorofila , Clorofila A , ADN Bacteriano/genética , Consorcios Microbianos/genética , Consorcios Microbianos/fisiología , Oxidorreductasas/genética , Oxidorreductasas/metabolismo , Océano Pacífico , Filogenia , ARN Ribosómico 16S/genética , Roseobacter/genética , Roseobacter/aislamiento & purificación , Roseobacter/metabolismo , Análisis de Secuencia de ADN , Sulfuros/metabolismo , Temperatura
19.
Comp Biochem Physiol B Biochem Mol Biol ; 166(2): 165-71, 2013 Oct.
Artículo en Inglés | MEDLINE | ID: mdl-23994361

RESUMEN

We purified D-amino acid oxidase (EC 1.4.3.3, DAO) from Xenopus laevis tadpoles. The optimal temperature and pH for enzyme activity were 35-40 °C and 8.3-9.0, respectively, depending on the substrate amino acids available to the enzyme; the highest activity was observed with D-proline followed by D-phenylalanine. Activity was significantly inhibited by p-hydroxymercuribenzoate, but only moderately by p-chloromercuribenzoate or benzoate. Enzyme activity was increased until the final tadpole stage, but was reduced to one-third in the adult and was localized primarily in the kidney. The tadpoles contained high concentrations of D-proline close to the final developmental stage and nearly no D-amino acids were detected in the adult frog, indicating that D-amino acid oxidase functions in metamorphosis.


Asunto(s)
D-Aminoácido Oxidasa/aislamiento & purificación , Larva/enzimología , Metamorfosis Biológica , Xenopus laevis/metabolismo , Aminoácidos , Animales , D-Aminoácido Oxidasa/antagonistas & inhibidores , D-Aminoácido Oxidasa/química , D-Aminoácido Oxidasa/metabolismo , Hidroximercuribenzoatos/farmacología , Larva/crecimiento & desarrollo , Larva/metabolismo , Fenilalanina/metabolismo , Fenilalanina/farmacología , Prolina/química , Prolina/farmacología , Especificidad por Sustrato , Xenopus laevis/crecimiento & desarrollo
20.
Mar Biotechnol (NY) ; 9(4): 429-36, 2007.
Artículo en Inglés | MEDLINE | ID: mdl-17497195

RESUMEN

The diversity and distribution of sulfate-reducing prokaryotes (SRP) was investigated in the Nankai Trough sediments of off-central Japan by exploring the diversity of a functional gene, dissimilatory sulfite reductase (dsrAB). Bulk DNAs were extracted from five piston-cored samples (up to 4.5 m long) with 41 vertical sections, and full-length dsrABgene sequences (ca. 1.9 kb) were PCR amplified and cloned. A total of 382 dsrAB clones yielded eight phylogenetic groups with an indigenous group forming a unique dsrAB lineage. The deltaproteobacterial dsrAB genes were found in almost all sediment samples, especially in the surface layer. One unique dsrAB clone group was also widespread in the dsrAB profiles of the studied sediments, and the percentage of its clones was generally shown gradual increase with sediment depth.


Asunto(s)
Variación Genética/genética , Sedimentos Geológicos/análisis , Hidrogenosulfito Reductasa/genética , Filogenia , Células Procariotas , Japón , Océanos y Mares , Reacción en Cadena de la Polimerasa
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