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1.
Exp Cell Res ; 383(1): 111494, 2019 10 01.
Artículo en Inglés | MEDLINE | ID: mdl-31306655

RESUMEN

Mesenchymal stem cells home towards inflammatory microenvironments, such as the tumour stroma, where they have been shown to have both pro- and anti-tumorigenic effects. Here, we demonstrate that the aspartic acid protease cathepsin D is part of the chemoattraction process. Using a Boyden chamber co-culture system, the migration of the mesenchymal stem cells and their invasion through Matrigel increased in the presence of breast cancer MDA-MB-231 cells, colon cancer HT29 cells or their conditioned media. Mesenchymal stem cell movement was reduced by protease inhibitors of matrix metalloproteinases and by pepstatin A, an inhibitor of cathepsin D. We confirmed a role for cathepsin D through addition of recombinant protein, upregulation of cathepsin D release using chloroquine and knockdown of cathepsin D expression. While all cell types expressed active cathepsin D, enzymatically inactive precursor procathepsin D was expressed only at low levels by mesenchymal stem cells. Expression in mesenchymal stem cells was increased following co-culture with cancer cells. The chemoattractive effect of cathepsin D required its enzymatic activity, but not changes in mesenchymal stem cell proliferation or adhesion rates. In conclusion, cathepsin D and its precursors enhance mesenchymal stem cell homing towards tumour sites, most likely by enzymatic mechanisms.


Asunto(s)
Neoplasias de la Mama/patología , Catepsina D/metabolismo , Movimiento Celular , Neoplasias del Colon/patología , Precursores Enzimáticos/metabolismo , Células Madre Mesenquimatosas/patología , Microambiente Tumoral , Neoplasias de la Mama/metabolismo , Adhesión Celular , Proliferación Celular , Células Cultivadas , Técnicas de Cocultivo , Colágeno , Neoplasias del Colon/metabolismo , Medios de Cultivo Condicionados , Combinación de Medicamentos , Femenino , Humanos , Laminina , Células Madre Mesenquimatosas/metabolismo , Proteoglicanos
2.
Gen Comp Endocrinol ; 292: 113443, 2020 06 01.
Artículo en Inglés | MEDLINE | ID: mdl-32097662

RESUMEN

We de novo assembled a transcriptome for early life-stages of the Aotearoa-New Zealand crayfish, Paranephrops zealandicus, establishing the first genetic resource for this under-developed aquaculture species and for the Paranephrops genus. Mining of this transcriptome for neuropeptides and their putative cognate G protein-coupled receptors (GPCRs) yielded a comprehensive catalogue of neuropeptides, but few putative neuropeptide GPCRs. Of the neuropeptides commonly identified from decapod transcriptomes, only crustacean female sex hormone and insulin-like peptide were absent from our trinity de novo transcriptome assembly, and also RNA-sequence reads. We identified 63 putative neuropeptide precursors from 43 families, predicted to yield 122 active peptides. Transcripts encoding 26 putative neuropeptide GPCRs were identified but were often incomplete. Putative GPCRs for 15 of the neuropeptides identified here were absent from our transcriptome and RNAseq reads. These data highlight the diverse neuropeptide systems already present at the early development life stages sampled here for P. zealandicus.


Asunto(s)
Astacoidea/embriología , Astacoidea/genética , Neuropéptidos/genética , Receptores Acoplados a Proteínas G/genética , Transcriptoma/genética , Secuencia de Aminoácidos , Animales , Embrión no Mamífero/metabolismo , Femenino , Regulación del Desarrollo de la Expresión Génica , Neuropéptidos/química , Neuropéptidos/metabolismo , Nueva Zelanda , Receptores Acoplados a Proteínas G/metabolismo , Transducción de Señal
3.
Nucleic Acids Res ; 46(4): 1927-1944, 2018 02 28.
Artículo en Inglés | MEDLINE | ID: mdl-29325104

RESUMEN

When a stop codon is at the 80S ribosomal A site, there are six nucleotides (+4 to +9) downstream that are inferred to be occupying the mRNA channel. We examined the influence of these downstream nucleotides on translation termination success or failure in mammalian cells at the three stop codons. The expected hierarchy in the intrinsic fidelity of the stop codons (UAA>UAG>>UGA) was observed, with highly influential effects on termination readthrough mediated by nucleotides at position +4 and position +8. A more complex influence was observed from the nucleotides at positions +5 and +6. The weakest termination contexts were most affected by increases or decreases in the concentration of the decoding release factor (eRF1), indicating that eRF1 binding to these signals was rate-limiting. When termination efficiency was significantly reduced by cognate suppressor tRNAs, the observed influence of downstream nucleotides was maintained. There was a positive correlation between experimentally measured signal strength and frequency of the signal in eukaryotic genomes, particularly in Saccharomyces cerevisiae and Drosophila melanogaster. We propose that termination efficiency is not only influenced by interrogation of the stop signal directly by the release factor, but also by downstream ribosomal interactions with the mRNA nucleotides in the entry channel.


Asunto(s)
Codón de Terminación , Terminación de la Cadena Péptídica Traduccional , ARN Mensajero/metabolismo , Animales , Células COS , Chlorocebus aethiops , Drosophila melanogaster/genética , Células HEK293 , Humanos , Nucleótidos/metabolismo , Factores de Terminación de Péptidos/metabolismo , ARN de Transferencia/metabolismo , Ribosomas/metabolismo , Saccharomyces cerevisiae/genética
4.
BMC Genomics ; 19(1): 832, 2018 Nov 21.
Artículo en Inglés | MEDLINE | ID: mdl-30463532

RESUMEN

BACKGROUND: Having conquered water surfaces worldwide, the semi-aquatic bugs occupy ponds, streams, lakes, mangroves, and even open oceans. The diversity of this group has inspired a range of scientific studies from ecology and evolution to developmental genetics and hydrodynamics of fluid locomotion. However, the lack of a representative water strider genome hinders our ability to more thoroughly investigate the molecular mechanisms underlying the processes of adaptation and diversification within this group. RESULTS: Here we report the sequencing and manual annotation of the Gerris buenoi (G. buenoi) genome; the first water strider genome to be sequenced thus far. The size of the G. buenoi genome is approximately 1,000 Mb, and this sequencing effort has recovered 20,949 predicted protein-coding genes. Manual annotation uncovered a number of local (tandem and proximal) gene duplications and expansions of gene families known for their importance in a variety of processes associated with morphological and physiological adaptations to a water surface lifestyle. These expansions may affect key processes associated with growth, vision, desiccation resistance, detoxification, olfaction and epigenetic regulation. Strikingly, the G. buenoi genome contains three insulin receptors, suggesting key changes in the rewiring and function of the insulin pathway. Other genomic changes affecting with opsin genes may be associated with wavelength sensitivity shifts in opsins, which is likely to be key in facilitating specific adaptations in vision for diverse water habitats. CONCLUSIONS: Our findings suggest that local gene duplications might have played an important role during the evolution of water striders. Along with these findings, the sequencing of the G. buenoi genome now provides us the opportunity to pursue exciting research opportunities to further understand the genomic underpinnings of traits associated with the extreme body plan and life history of water striders.


Asunto(s)
Genoma , Heterópteros/genética , Heterópteros/fisiología , Proteínas de Insectos/genética , Adaptación Fisiológica , Animales , Evolución Molecular , Genómica , Heterópteros/clasificación , Fenotipo , Filogenia
5.
Biochem Soc Trans ; 46(6): 1615-1625, 2018 12 17.
Artículo en Inglés | MEDLINE | ID: mdl-30420414

RESUMEN

Translational stop codons, UAA, UAG, and UGA, form an integral part of the universal genetic code. They are of significant interest today for their underlying fundamental role in terminating protein synthesis, but also for their potential utilisation for programmed alternative translation events. In diverse organisms, UAA has wide usage, but it is puzzling that the high fidelity UAG is selected against and yet UGA, vulnerable to suppression, is widely used, particularly in those archaeal and bacterial genomes with a high GC content. In canonical protein synthesis, stop codons are interpreted by protein release factors that structurally and functionally mimic decoding tRNAs and occupy the decoding site on the ribosome. The release factors make close contact with the decoding complex through multiple interactions. Correct interactions cause conformational changes resulting in new and enhanced contacts with the ribosome, particularly between specific bases in the mRNA and rRNA. The base following the stop codon (fourth or +4 base) may strongly influence decoding efficiency, facilitating alternative non-canonical events like frameshifting or selenocysteine incorporation. The fourth base is drawn into the decoding site with a compacted stop codon in the eukaryotic termination complex. Surprisingly, mRNA sequences upstream and downstream of this core tetranucleotide signal have a significant influence on the strength of the signal. Since nine bases downstream of the stop codon are within the mRNA channel, their interactions with rRNA, and r-proteins may affect efficiency. With this understanding, it is now possible to design stop signals of desired strength for specific applied purposes.


Asunto(s)
Codón de Terminación/genética , Ribosomas/metabolismo , Regulación de la Expresión Génica , Biosíntesis de Proteínas , ARN Mensajero/genética
6.
Electrophoresis ; 39(5-6): 824-832, 2018 03.
Artículo en Inglés | MEDLINE | ID: mdl-29125656

RESUMEN

Polyacrylamide gel electrophoresis is a universal tool in a biochemist's toolkit for protein and nucleic acid separation and subsequent visualisation and analysis. The standard formulation of polyacrylamide gels consists of acrylamide (ACM) monomer crosslinked with bisacrylamide (MBA) which creates a gel with excellent sieving properties but which is mechanically fragile and prone to tearing during post-electrophoresis manipulations involved in visualisation and analysis. By adding a poly(ethylene oxide) macro-crosslinker to the standard gel formulation, we have created a tough gel matrix that can be used to fractionate proteins and nucleic acids by polyacrylamide gel electrophoresis. The protein and nucleic acid resolving capabilities and performance during staining and electroblotting of the tough gel matrix rivals that of conventional acrylamide/bisacrylamide gels. The tough gel matrix is resistant to tear and remarkably elastic, capable of stretching to over four times its original length before breaking, and represents a significant improvement over standard polyacrylamide gel formulations.


Asunto(s)
Resinas Acrílicas/química , Electroforesis en Gel de Poliacrilamida/métodos , Fenómenos Mecánicos , Ácidos Nucleicos/análisis , Proteínas/análisis , Animales , Reactivos de Enlaces Cruzados/química , Immunoblotting/métodos , Polietilenglicoles/química , Polimerizacion , Spodoptera/química , Coloración y Etiquetado
7.
Environ Sci Technol ; 52(10): 6009-6022, 2018 05 15.
Artículo en Inglés | MEDLINE | ID: mdl-29634279

RESUMEN

Hyalella azteca is a cryptic species complex of epibenthic amphipods of interest to ecotoxicology and evolutionary biology. It is the primary crustacean used in North America for sediment toxicity testing and an emerging model for molecular ecotoxicology. To provide molecular resources for sediment quality assessments and evolutionary studies, we sequenced, assembled, and annotated the genome of the H. azteca U.S. Lab Strain. The genome quality and completeness is comparable with other ecotoxicological model species. Through targeted investigation and use of gene expression data sets of H. azteca exposed to pesticides, metals, and other emerging contaminants, we annotated and characterized the major gene families involved in sequestration, detoxification, oxidative stress, and toxicant response. Our results revealed gene loss related to light sensing, but a large expansion in chemoreceptors, likely underlying sensory shifts necessary in their low light habitats. Gene family expansions were also noted for cytochrome P450 genes, cuticle proteins, ion transporters, and include recent gene duplications in the metal sequestration protein, metallothionein. Mapping of differentially expressed transcripts to the genome significantly increased the ability to functionally annotate toxicant responsive genes. The H. azteca genome will greatly facilitate development of genomic tools for environmental assessments and promote an understanding of how evolution shapes toxicological pathways with implications for environmental and human health.


Asunto(s)
Anfípodos , Contaminantes Químicos del Agua , Animales , Ecotoxicología , Sedimentos Geológicos , América del Norte , Pruebas de Toxicidad
8.
Ann Bot ; 117(5): 833-43, 2016 04.
Artículo en Inglés | MEDLINE | ID: mdl-27013176

RESUMEN

BACKGROUND: The remarkable similarity of animal embryos at particular stages of development led to the proposal of a developmental hourglass. In this model, early events in development are less conserved across species but lead to a highly conserved 'phylotypic period'. Beyond this stage, the model suggests that development once again becomes less conserved, leading to the diversity of forms. Recent comparative studies of gene expression in animal groups have provided strong support for the hourglass model. How and why might such an hourglass pattern be generated? More importantly, how might early acting events in development evolve while still maintaining a later conserved stage? SCOPE: The discovery that an hourglass pattern may also exist in the embryogenesis of plants provides comparative data that may help us explain this phenomenon. Whether the developmental hourglass occurs in plants, and what this means for our understanding of embryogenesis in plants and animals is discussed. Models by which conserved early-acting genes might change their functional role in the evolution of gene networks, how networks buffer these changes, and how that might constrain, or confer diversity, of the body plan are also discused. CONCLUSIONS: Evidence of a morphological and molecular hourglass in plant and animal embryogenesis suggests convergent evolution. This convergence is likely due to developmental constraints imposed upon embryogenesis by the need to produce a viable embryo with an established body plan, controlled by the architecture of the underlying gene regulatory networks. As the body plan is largely laid down during the middle phases of embryo development in plants and animals, then it is perhaps not surprising this stage represents the narrow waist of the hourglass where the gene regulatory networks are the oldest and most robust and integrated, limiting species diversity and constraining morphological space.


Asunto(s)
Desarrollo Embrionario , Semillas , Animales , Flores/crecimiento & desarrollo , Regulación del Desarrollo de la Expresión Génica , Modelos Biológicos , Semillas/crecimiento & desarrollo
9.
Anal Biochem ; 447: 82-9, 2014 Feb 15.
Artículo en Inglés | MEDLINE | ID: mdl-24176934

RESUMEN

This work describes a quick semi-quantitative colony immunoassay (QSCI) method for immunoblot detection of intracellularly expressed proteins in both yeast and bacterial cells. After induction of protein expression, only 4.5 h is required for cell breakage, protein detection, and data analysis. This protocol was used to screen and unambiguously identify Saccharomyces cerevisiae cells efficiently overexpressing glutathione S-transferase (GST)-tagged Yih1 in addition to cells expressing the myc-tagged large 297-kDa Gcn1 protein. In addition, the method was used to identify Escherichia coli cells efficiently expressing His6-tagged Yih1 and a GST-tagged Gcn1 fragment, respectively. The protocol allows the use of both epitope-specific and protein-specific antibodies. The same colony immunoassay can also be used to determine the minimal concentration of inducing agent sufficient for induction of optimal protein expression (e.g., galactose for yeast, isopropyl ß-D-1-thiogalactopyranoside [IPTG] for E. coli). To our knowledge, this is the first report on a rapid low-cost procedure that allows the calibration of inducing agent on solid medium.


Asunto(s)
Escherichia coli/genética , Inmunoensayo/métodos , Proteínas Recombinantes/genética , Saccharomyces cerevisiae/genética , Escherichia coli/citología , Expresión Génica , Ingeniería Genética , Saccharomyces cerevisiae/citología
10.
J Ind Microbiol Biotechnol ; 40(10): 1105-16, 2013 Oct.
Artículo en Inglés | MEDLINE | ID: mdl-23907252

RESUMEN

Listeria monocytogenes is a food-borne pathogen which causes listeriosis and is difficult to eradicate from seafood processing environments; therefore, more effective control methods need to be developed. This study investigated the effectiveness of three bacteriophages (LiMN4L, LiMN4p and LiMN17), individually or as a three-phage cocktail at ≈9 log10 PFU/ml, in the lysis of three seafood-borne L. monocytogenes strains (19CO9, 19DO3 and 19EO3) adhered to a fish broth layer on stainless steel coupon (FBSSC) and clean stainless steel coupon (SSC), in 7-day biofilm, and dislodged biofilm cells at 15 ± 1 °C. Single phage treatments (LiMN4L, LiMN4p or LiMN17) decreased bacterial cells adhered to FBSSC and SSC by ≈3-4.5 log units. Phage cocktail reduced the cells on both surfaces (≈3.8-4.5 and 4.6-5.4 log10 CFU/cm², respectively), to less than detectable levels after ≈75 min (detection limit = 0.9 log10 CFU/cm²). The phage cocktail at ≈5.8, 6.5 and 7.5 log10 PFU/cm² eliminated Listeria contamination (≈1.5-1.7 log10 CFU/cm²) on SSC in ≈15 min. One-hour phage treatments (LiMN4p, LiMN4L and cocktail) in three consecutive applications resulted in a decrease of 7-day L. monocytogenes biofilms (≈4 log10 CFU/cm²) by ≈2-3 log units. Single phage treatments reduced dislodged biofilm cells of each L. monocytogenes strain by ≈5 log10 CFU/ml in 1 h. The three phages were effective in controlling L. monocytogenes on stainless steel either clean or soiled with fish proteins which is likely to occur in seafood processing environments. Phages were more effective on biofilm cells dislodged from the surface compared with undisturbed biofilm cells. Therefore, for short-term phage treatments of biofilm it should be considered that some disruption of the biofilm cells from the surface prior to phage application will be required.


Asunto(s)
Bacteriófagos/fisiología , Biopelículas , Descontaminación/métodos , Contaminación de Alimentos/prevención & control , Listeria monocytogenes/fisiología , Listeria monocytogenes/virología , Alimentos Marinos/microbiología , Animales , Adhesión Bacteriana , Recuento de Colonia Microbiana , Proteínas de Peces/análisis , Contaminación de Alimentos/análisis , Listeria monocytogenes/aislamiento & purificación , Acero Inoxidable/análisis
11.
Biol Rev Camb Philos Soc ; 98(4): 1003-1015, 2023 08.
Artículo en Inglés | MEDLINE | ID: mdl-36808687

RESUMEN

Forestry management worldwide has become increasingly effective at obtaining high timber yields from productive forests. In New Zealand, a focus on improving an increasingly successful and largely Pinus radiata plantation forestry model over the last 150 years has resulted in some of the most productive timber forests in the temperate zone. In contrast to this success, the full range of forested landscapes across New Zealand, including native forests, are impacted by an array of pressures from introduced pests, diseases, and a changing climate, presenting a collective risk of losses in biological, social and economic value. As the national government policies incentivise reforestation and afforestation, the social acceptability of some forms of newly planted forests is also being challenged. Here, we review relevant literature in the area of integrated forest landscape management to optimise forests as nature-based solutions, presenting 'transitional forestry' as a model design and management paradigm appropriate to a range of forest types, where forest purpose is placed at the heart of decision making. We use New Zealand as a case study region, describing how this purpose-led transitional forestry model can benefit a cross section of forest types, from industrialised forest plantations to dedicated conservation forests and a range of multiple-purpose forests in between. Transitional forestry is an ongoing multi-decade process of change from current 'business-as-usual' forest management to future systems of forest management, embedded across a continuum of forest types. This holistic framework incorporates elements to enhance efficiencies of timber production, improve overall forest landscape resilience, and reduce some potential negative environmental impacts of commercial plantation forestry, while allowing the ecosystem functioning of commercial and non-commercial forests to be maximised, with increased public and biodiversity conservation value. Implementation of transitional forestry addresses tensions that arise between meeting climate mitigation targets and improving biodiversity criteria through afforestation, alongside increasing demand for forest biomass feedstocks to meet the demands of near-term bioenergy and bioeconomy goals. As ambitious government international targets are set for reforestation and afforestation using both native and exotic species, there is an increasing opportunity to make such transitions via integrated thinking that optimises forest values across a continuum of forest types, while embracing the diversity of ways in which such targets can be reached.


Asunto(s)
Ecosistema , Agricultura Forestal , Nueva Zelanda , Bosques , Biodiversidad , Conservación de los Recursos Naturales , Árboles
12.
Nucleic Acids Res ; 38(22): 8039-50, 2010 Dec.
Artículo en Inglés | MEDLINE | ID: mdl-20705650

RESUMEN

eIF4E-binding proteins (4E-BPs) regulate translation of mRNAs in eukaryotes. However the extent to which specific mRNA targets are regulated by 4E-BPs remains unknown. We performed translational profiling by microarray analysis of polysome and monosome associated mRNAs in wild-type and mutant cells to identify mRNAs in yeast regulated by the 4E-BPs Caf20p and Eap1p; the first-global comparison of 4E-BP target mRNAs. We find that yeast 4E-BPs modulate the translation of >1000 genes. Most target mRNAs differ between the 4E-BPs revealing mRNA specificity for translational control by each 4E-BP. This is supported by observations that eap1Δ and caf20Δ cells have different nitrogen source utilization defects, implying different mRNA targets. To account for the mRNA specificity shown by each 4E-BP, we found correlations between our data sets and previously determined targets of yeast mRNA-binding proteins. We used affinity chromatography experiments to uncover specific RNA-stabilized complexes formed between Caf20p and Puf4p/Puf5p and between Eap1p and Puf1p/Puf2p. Thus the combined action of each 4E-BP with specific 3'-UTR-binding proteins mediates mRNA-specific translational control in yeast, showing that this form of translational control is more widely employed than previously thought.


Asunto(s)
Factores Eucarióticos de Iniciación/fisiología , Regulación Fúngica de la Expresión Génica , Biosíntesis de Proteínas , Proteínas de Unión al ARN/metabolismo , Proteínas de Saccharomyces cerevisiae/fisiología , Saccharomyces cerevisiae/genética , Factores de Transcripción/fisiología , Aminoácidos/metabolismo , Ciclinas/genética , Factores Eucarióticos de Iniciación/genética , Eliminación de Gen , Nitrógeno/metabolismo , Análisis de Secuencia por Matrices de Oligonucleótidos , ARN Mensajero/metabolismo , Saccharomyces cerevisiae/crecimiento & desarrollo , Saccharomyces cerevisiae/metabolismo , Proteínas de Saccharomyces cerevisiae/genética , Factores de Transcripción/genética , Transcripción Genética
13.
Sci Rep ; 11(1): 3898, 2021 02 16.
Artículo en Inglés | MEDLINE | ID: mdl-33594128

RESUMEN

Glacier ice is an extreme environment in which most animals cannot survive. Here we report the colonization of high elevation, climate-threatened glaciers along New Zealand's southwestern coast by species of Arthropoda, Nematoda, Platyhelminthes, Rotifera and Tardigrada. Based on DNA barcoding and haplotype-inferred evidence for deep genetic variability, at least 12 undescribed species are reported, some of which have persisted in this niche habitat throughout the Pleistocene. These findings identify not only an atypical biodiversity hotspot but also highlight the adaptive plasticity of microinvertebrate Animalia.

14.
Genome Biol Evol ; 12(7): 1099-1188, 2020 07 01.
Artículo en Inglés | MEDLINE | ID: mdl-32442304

RESUMEN

The tremendous diversity of Hymenoptera is commonly attributed to the evolution of parasitoidism in the last common ancestor of parasitoid sawflies (Orussidae) and wasp-waisted Hymenoptera (Apocrita). However, Apocrita and Orussidae differ dramatically in their species richness, indicating that the diversification of Apocrita was promoted by additional traits. These traits have remained elusive due to a paucity of sawfly genome sequences, in particular those of parasitoid sawflies. Here, we present comparative analyses of draft genomes of the primarily phytophagous sawfly Athalia rosae and the parasitoid sawfly Orussus abietinus. Our analyses revealed that the ancestral hymenopteran genome exhibited traits that were previously considered unique to eusocial Apocrita (e.g., low transposable element content and activity) and a wider gene repertoire than previously thought (e.g., genes for CO2 detection). Moreover, we discovered that Apocrita evolved a significantly larger array of odorant receptors than sawflies, which could be relevant to the remarkable diversification of Apocrita by enabling efficient detection and reliable identification of hosts.


Asunto(s)
Especiación Genética , Genoma de los Insectos , Interacciones Huésped-Parásitos/genética , Himenópteros/genética , Secuencia de Aminoácidos , Animales , Secuencia Conservada , Elementos Transponibles de ADN , Femenino , Dosificación de Gen , Glicoproteínas/genética , Herbivoria/genética , Inmunidad/genética , Proteínas de Insectos/genética , Masculino , Familia de Multigenes , Receptores Odorantes/genética , Conducta Social , Visión Ocular/genética
15.
Genome Biol ; 20(1): 64, 2019 04 02.
Artículo en Inglés | MEDLINE | ID: mdl-30935422

RESUMEN

BACKGROUND: The Hemiptera (aphids, cicadas, and true bugs) are a key insect order, with high diversity for feeding ecology and excellent experimental tractability for molecular genetics. Building upon recent sequencing of hemipteran pests such as phloem-feeding aphids and blood-feeding bed bugs, we present the genome sequence and comparative analyses centered on the milkweed bug Oncopeltus fasciatus, a seed feeder of the family Lygaeidae. RESULTS: The 926-Mb Oncopeltus genome is well represented by the current assembly and official gene set. We use our genomic and RNA-seq data not only to characterize the protein-coding gene repertoire and perform isoform-specific RNAi, but also to elucidate patterns of molecular evolution and physiology. We find ongoing, lineage-specific expansion and diversification of repressive C2H2 zinc finger proteins. The discovery of intron gain and turnover specific to the Hemiptera also prompted the evaluation of lineage and genome size as predictors of gene structure evolution. Furthermore, we identify enzymatic gains and losses that correlate with feeding biology, particularly for reductions associated with derived, fluid nutrition feeding. CONCLUSIONS: With the milkweed bug, we now have a critical mass of sequenced species for a hemimetabolous insect order and close outgroup to the Holometabola, substantially improving the diversity of insect genomics. We thereby define commonalities among the Hemiptera and delve into how hemipteran genomes reflect distinct feeding ecologies. Given Oncopeltus's strength as an experimental model, these new sequence resources bolster the foundation for molecular research and highlight technical considerations for the analysis of medium-sized invertebrate genomes.


Asunto(s)
Evolución Molecular , Genoma de los Insectos , Hemípteros/genética , Secuencia de Aminoácidos , Animales , Dedos de Zinc CYS2-HIS2 , Conducta Alimentaria , Dosificación de Gen , Perfilación de la Expresión Génica , Transferencia de Gen Horizontal , Genes Homeobox , Hemípteros/crecimiento & desarrollo , Hemípteros/metabolismo , Pigmentación/genética , Olfato , Factores de Transcripción/genética
16.
Nucleic Acids Res ; 34(7): 1959-73, 2006.
Artículo en Inglés | MEDLINE | ID: mdl-16614446

RESUMEN

Six diverse prokaryotic and five eukaryotic genomes were compared to deduce whether the protein synthesis termination signal has common determinants within and across both kingdoms. Four of the six prokaryotic and all of the eukaryotic genomes investigated demonstrated a similar pattern of nucleotide bias both 5' and 3' of the stop codon. A preferred core signal of 4 nt was evident, encompassing the stop codon and the following nucleotide. Codons decoded by hyper-modified tRNAs were over-represented in the region 5' to the stop codon in genes from both kingdoms. The origin of the 3' bias was more variable particularly among the prokaryotic organisms. In both kingdoms, genes with the highest expression index exhibited a strong bias but genes with the lowest expression showed none. Absence of bias in parasitic prokaryotes may reflect an absence of pressure to evolve more efficient translation. Experiments were undertaken to determine if a correlation existed between bias in signal abundance and termination efficiency. In Escherichia coli signal abundance correlated with termination efficiency for UAA and UGA stop codons, but not in mammalian cells. Termination signals that were highly inefficient could be made more efficient by increasing the concentration of the cognate decoding release factor.


Asunto(s)
Codón de Terminación , Células Eucariotas/metabolismo , Terminación de la Cadena Péptídica Traduccional , Células Procariotas/metabolismo , Animales , Células COS , Chlorocebus aethiops , Biología Computacional , Genómica , Humanos , Nucleótidos/análisis , Factores de Terminación de Péptidos/metabolismo , ARN de Transferencia/metabolismo
17.
Int J Oncol ; 52(2): 579-588, 2018 Feb.
Artículo en Inglés | MEDLINE | ID: mdl-29207190

RESUMEN

Prostate cancer is initially sensitive to hormone therapy; however, over time the majority of patients progress to a hormone-insensitive form classified as castration-resistant prostate cancer (CRPC). CRPC is highly metastatic and patients have a poor prognosis. Thus, new drugs for the treatment of this disease are required. In this study, we therefore examined the cytotoxic effects and anticancer mechanism(s) of action of second generation curcumin analogs towards CRPC cells. For this purpose, PC3 and DU145 cells were treated with a series of curcumin analogs at 0-10 µM for 72 h and cytotoxicity was determined by the sulforhodamine B (SRB) assay. Two compounds, 1-isopropyl-3,5-bis(pyridin-3-ylmethylene)-4-piperidone (RL118) and 1-methyl-3,5-[(6-methoxynaphthalen-2-yl)methylene]-4-piperidone (RL121), were found to have the most potent cytotoxic effect with EC50 values of 0.50 and 0.58 µM in the PC3 cells and EC50 values of 0.76 and 0.69 µM in the DU145 cells, respectively. Thus, further experiments were performed focusing on these two compounds. Flow cytometry was performed to determine their effects on the cell cycle and apoptosis. Both analogs increased the number of cells in the G2/M phase of the cell cycle and induced apoptosis. Specifically, in the PC3 cells, RL121 increased the number of cells in the G2/M phase by 86% compared to the control, while RL118 increased the number of cells in the G2/M phase by 42% compared to the control after 24 h. Moreover, both RL118 and RL121 induced the apoptosis of both cell lines. In the DU145 cells, a 38-fold increase in the number of apoptotic cells was elicited by RL118 and a 78-fold increase by RL121 compared to the control. Furthermore, the effects of both analogs on the expression of key proteins involved in cell proliferation were also determined by western blot analysis. The results revealed that both analogs inhibited the expression of nuclear factor (NF)-κB (p65/RelA), eukaryotic translation initiation factor 4E-binding protein 1 (4E-BP1), p-4E-BP1, mammalian target of rapamycin (mTOR), p-mTOR, AKT and p-AKT. Thus, the findings of this study provide evidence that RL118 and RL121 have potent anticancer activity against CPRC cells, and both analogs warrant further investigation in vivo.


Asunto(s)
Antineoplásicos/farmacología , Apoptosis/efectos de los fármacos , Curcumina/análogos & derivados , Curcumina/farmacología , Neoplasias de la Próstata Resistentes a la Castración , Línea Celular Tumoral , Humanos , Masculino , Piperidonas/farmacología , Piridinas/farmacología
19.
Mol Cell Biol ; 35(3): 544-54, 2015 Feb.
Artículo en Inglés | MEDLINE | ID: mdl-25421713

RESUMEN

The spatial organization of eukaryotic genomes is linked to their functions. However, how individual features of the global spatial structure contribute to nuclear function remains largely unknown. We previously identified a high-frequency interchromosomal interaction within the Saccharomyces cerevisiae genome that occurs between the intergenic spacer of the ribosomal DNA (rDNA) repeats and the intergenic sequence between the locus encoding the second largest RNA polymerase I subunit and a lysine tRNA gene [i.e., RPA135-tK(CUU)P]. Here, we used quantitative chromosome conformation capture in combination with replacement mapping to identify a 75-bp sequence within the RPA135-tK(CUU)P intergenic region that is involved in the interaction. We demonstrate that the RPA135-IGS1 interaction is dependent on the rDNA copy number and the Msn2 protein. Surprisingly, we found that the interaction does not govern RPA135 transcription. Instead, replacement of a 605-bp region within the RPA135-tK(CUU)P intergenic region results in a reduction in the RPA135-IGS1 interaction level and fluctuations in rDNA copy number. We conclude that the chromosomal interaction that occurs between the RPA135-tK(CUU)P and rDNA IGS1 loci stabilizes rDNA repeat number and contributes to the maintenance of nucleolar stability. Our results provide evidence that the DNA loci involved in chromosomal interactions are composite elements, sections of which function in stabilizing the interaction or mediating a functional outcome.


Asunto(s)
ADN Ribosómico/genética , Regulación Fúngica de la Expresión Génica/genética , Genes de ARNr/genética , ARN Polimerasa I/genética , Proteínas de Saccharomyces cerevisiae/metabolismo , Saccharomyces cerevisiae/genética , Animales , Cromosomas Fúngicos/genética , ADN de Hongos/genética , Sitios Genéticos , ARN Polimerasa I/metabolismo , Transcripción Genética/genética
20.
Nutrients ; 7(3): 1787-97, 2015 Mar 11.
Artículo en Inglés | MEDLINE | ID: mdl-25768950

RESUMEN

Many insects are capable of remarkable changes in biology and form in response to their environment or diet. The most extreme example of these are polyphenisms, which are when two or more different phenotypes are produced from a single genotype in response to the environment. Polyphenisms provide a fascinating opportunity to study how the environment affects an animal's genome, and how this produces changes in form. Here we review the current state of knowledge of the molecular basis of polyphenisms and what can be learnt from them to understand how nutrition may influence our own genomes.


Asunto(s)
Dieta , Epigénesis Genética , Epigenómica , Genotipo , Insectos/genética , Estado Nutricional/genética , Fenotipo , Animales , Ambiente , Pleiotropía Genética , Humanos
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