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Displaying Results 1 - 25 of 142 on page 1 of 6
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A cenp-s/x complex assembles at the centromere in s and g2 phases of the human cell cycle
(2018)
Dornblut, C.; Quinn, N.; Monajambashi, S.; Prendergast, L.; van Vuuren, C.; Munch, S.; ...
A cenp-s/x complex assembles at the centromere in s and g2 phases of the human cell cycle
(2018)
Dornblut, C.; Quinn, N.; Monajambashi, S.; Prendergast, L.; van Vuuren, C.; Munch, S.; Deng, W.; Leonhardt, H.; Cardoso, M. C.; Hoischen, C.; Diekmann, S.; Sullivan, K. F.
Abstract:
The functional identity of centromeres arises from a set of specific nucleoprotein particle subunits of the centromeric chromatin fibre. These include CENP-A and histone H3 nucleosomes and a novel nucleosome-like complex of CENPs -T, -W, -S and -X. Fluorescence cross-correlation spectroscopy and Forster resonance energy transfer (FRET) revealed that human CENP-S and -X exist principally in complex in soluble form and retain proximity when assembled at centromeres. Conditional labelling experiments show that they both assemble de novo during S phase and G2, increasing approximately three-to fourfold in abundance at centromeres. Fluorescence recovery after photobleaching (FRAP) measurements documented steady-state exchange between soluble and assembled pools, with CENP-X exchanging approximately 10 times faster than CENP-S (t(1/2) similar to 10 min versus 120 min). CENP-S binding to sites of DNA damage was quite distinct, with a FRAP half-time of approximately 160 s. Fluorescent two-h...
http://hdl.handle.net/10379/11239
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A fluorescence anisotropy method for measuring protein concentration in complex cell culture media
(2014)
Groza, RC,Calvet, A,Ryder, AG; Groza, Radu Constantin; Calvet, Amandine; Ryder, Alan G.
A fluorescence anisotropy method for measuring protein concentration in complex cell culture media
(2014)
Groza, RC,Calvet, A,Ryder, AG; Groza, Radu Constantin; Calvet, Amandine; Ryder, Alan G.
Abstract:
Journal article
The rapid, quantitative analysis of the complex cell culture media used in biopharmaceutical manufacturing is of critical importance. Requirements for cell culture media composition profiling, or changes in specific analyte concentrations (e. g. amino acids in the media or product protein in the bioprocess broth) often necessitate the use of complicated analytical methods and extensive sample handling. Rapid spectroscopic methods like multi-dimensional fluorescence (MDF) spectroscopy have been successfully applied for the routine determination of compositional changes in cell culture media and bioprocess broths. Quantifying macromolecules in cell culture media is a specific challenge as there is a need to implement measurements rapidly on the prepared media. However, the use of standard fluorescence spectroscopy is complicated by the emission overlap from many media components. Here, we demonstrate how combining anisotropy measurements with standard total synchro...
http://hdl.handle.net/10379/4521
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A fluorescence anisotropy method for measuring protein concentration in complex cell culture media
(2018)
Groza, Radu Constantin; Calvet, Amandine; Ryder, Alan G.
A fluorescence anisotropy method for measuring protein concentration in complex cell culture media
(2018)
Groza, Radu Constantin; Calvet, Amandine; Ryder, Alan G.
http://hdl.handle.net/10379/11741
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A high through-put screen for small molecules modulating mcm2 phosphorylation identifies ryuvidine as an inducer of the dna damage response
(2018)
FitzGerald, Jennifer; Murillo, Laura S.; O'Brien, Gemma; O'Connell, Enda; O...
A high through-put screen for small molecules modulating mcm2 phosphorylation identifies ryuvidine as an inducer of the dna damage response
(2018)
FitzGerald, Jennifer; Murillo, Laura S.; O'Brien, Gemma; O'Connell, Enda; O'Connor, Aisling; Wu, Kevin; Wang, Guan-Nan; Rainey, Michael D.; Natoni, Alessandro; Healy, Sandra; O'Dwyer, Michael; Santocanale, Corrado
Abstract:
DNA replication is an essential process for cell division and as such it is a process that is directly targeted by several anticancer drugs. CDC7 plays an essential role in the activation of replication origins and has recently been proposed as a novel target for drug discovery. The MCM DNA helicase complex (MCM2-7) is a key target of the CDC7 kinase, and MCM phosphorylation status at specific sites is a reliable biomarker of CDC7 cellular activity. In this work we describe a cell-based assay that utilizes the "In Cell Western Technique'' (ICW) to identify compounds that affect cellular CDC7 activity. By screening a library of approved drugs and kinase inhibitors we found several compounds that can affect CDC7-dependent phosphorylation of MCM2 in HeLa cells. Among these, Mitoxantrone, a topoisomerase inhibitor, and Ryuvidine, previously described as a CDK4 inhibitor, cause a reduction in phosphorylated MCM2 levels and a sudden blockade of DNA synthesis that is...
http://hdl.handle.net/10379/11484
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A profusion of upstream open reading frame mechanisms in polyamine-responsive translational regulation
(2010)
Ivanov, Ivaylo P.; Atkins, John F.; Michael, Antony J.
A profusion of upstream open reading frame mechanisms in polyamine-responsive translational regulation
(2010)
Ivanov, Ivaylo P.; Atkins, John F.; Michael, Antony J.
Abstract:
In many eukaryotic mRNAs one or more short 'upstream' open reading frames, uORFs, precede the initiator of the main coding sequence. Upstream ORFs are functionally diverse as illustrated by their variety of features in polyamine pathway biosynthetic mRNAs. Their propensity to act as sensors for regulatory circuits and to amplify the signals likely explains their occurrence in most polyamine pathway mRNAs. The uORF-mediated polyamine responsive autoregulatory circuits found in polyamine pathway mRNAs exemplify the translationally regulated dynamic interface between components of the proteome and metabolism.
http://hdl.handle.net/10468/5028
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A role for the p53 tumour suppressor in regulating the balance between homologous recombination and non-homologous end joining
(2018)
Moureau, Sylvie; Luessing, Janna; Harte, Emma Christina; Voisin, Muriel; Lowndes, Noel ...
A role for the p53 tumour suppressor in regulating the balance between homologous recombination and non-homologous end joining
(2018)
Moureau, Sylvie; Luessing, Janna; Harte, Emma Christina; Voisin, Muriel; Lowndes, Noel Francis
Abstract:
Loss of p53, a transcription factor activated by cellular stress, is a frequent event in cancer. The role of p53 in tumour suppression is largely attributed to cell fate decisions. Here, we provide evidence supporting a novel role for p53 in the regulation of DNA double-strand break (DSB) repair pathway choice. 53BP1, another tumour suppressor, was initially identified as p53 Binding Protein 1, and has been shown to inhibit DNA end resection, thereby stimulating non-homologous end joining (NHEJ). Yet another tumour suppressor, BRCA1, reciprocally promotes end resection and homologous recombination (HR). Here, we show that in both human and mouse cells, the absence of p53 results in impaired 53BP1 focal recruitment to sites of DNA damage induced by ionizing radiation. This effect is largely independent of cell cycle phase and the extent of DNA damage. In p53-deficient cells, diminished localization of 53BP1 is accompanied by a reciprocal increase in BRCA1 recruitment to DSBs. Consist...
http://hdl.handle.net/10379/12998
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Abnormal centrosomal structure and duplication in cep135-deficient vertebrate cells
(2018)
Inanc, B.; Putz, M.; Lalor, P.; Dockery, P.; Kuriyama, R.; Gergely, F.; Morrison, C. G.
Abnormal centrosomal structure and duplication in cep135-deficient vertebrate cells
(2018)
Inanc, B.; Putz, M.; Lalor, P.; Dockery, P.; Kuriyama, R.; Gergely, F.; Morrison, C. G.
Abstract:
Centrosomes are key microtubule-organizing centers that contain a pair of centrioles, conserved cylindrical, microtubule-based structures. Centrosome duplication occurs once per cell cycle and relies on templated centriole assembly. In many animal cells this process starts with the formation of a radially symmetrical cartwheel structure. The centrosomal protein Cep135 localizes to this cartwheel, but its role in vertebrates is not well understood. Here we examine the involvement of Cep135 in centriole function by disrupting the Cep135 gene in the DT40 chicken B-cell line. DT40 cells that lack Cep135 are viable and show no major defects in centrosome composition or function, although we note a small decrease in centriole numbers and a concomitant increase in the frequency of monopolar spindles. Furthermore, electron microscopy reveals an atypical structure in the lumen of Cep135-deficient centrioles. Centrosome amplification after hydroxyurea treatment increases significantly in Cep1...
http://hdl.handle.net/10379/12037
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Adolescent Risperidone treatment alters protein expression associated with protein trafficking and cellular metabolism in the adult rat prefrontal cortex.
(2014)
Farrelly, Lorna; Dicker, Patrick; Wynne, Kieran; English, Jane A; Cagney, Gerard; Föcki...
Adolescent Risperidone treatment alters protein expression associated with protein trafficking and cellular metabolism in the adult rat prefrontal cortex.
(2014)
Farrelly, Lorna; Dicker, Patrick; Wynne, Kieran; English, Jane A; Cagney, Gerard; Föcking, Melanie; Cotter, David R
Abstract:
<p><strong><em>This is the pre-peer reviewed version of the following article: </em></strong>Farrelly LA, Dicker P, Wynne K, English J, Cagney G, Föcking M, Cotter DR. Adolescent Risperidone treatment alters protein expression associated with protein trafficking and cellular metabolism in the adult rat prefrontal cortex. Proteomics. 2014;14(12):1574-8. <strong><em>which has been published in final form at </em>DOI: </strong>10.1002/pmic.201300466<strong><em>This article may be used for non-commercial purposes in accordance with <a href="http://olabout.wiley.com/WileyCDA/Section/id-828039.html#terms">Wiley Terms and Conditions for Self-Archiving</a>.</em></strong></p>
<p>The prefrontal cortex (PFC) is associated with mental health illnesses including schizophrenia, depression, bipolar disorder, and autism spectrum disorders. It richly expresses neuroreceptors which are ...
https://epubs.rcsi.ie/psychart/65
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Alternative uses for co-products: Harnessing the potential of valuable compounds from meat processing chains
(2017)
Mullen, Anne Maria; Álvarez García, Carlos; Zeugolis, Dimitrios; Henchion, Maeve; O...
Alternative uses for co-products: Harnessing the potential of valuable compounds from meat processing chains
(2017)
Mullen, Anne Maria; Álvarez García, Carlos; Zeugolis, Dimitrios; Henchion, Maeve; O'Neill, Eileen; Drummond, Liana
Abstract:
Opportunities for exploiting the inherent value of protein-rich meat processing co-products, in the context of increased global demand for protein and for sustainable processing systems, are discussed. While direct consumption maybe the most profitable route for some, this approach is influenced greatly by local and cultural traditions. A more profitable and sustainable approach may be found in recognizing this readily available and under-utilised resource can provide high value components, such as proteins, with targeted high value functionality of relevance to a variety of sectors. Applications in food & beverages, petfood biomedical and nutrition arenas are discussed. Utilization of the raw material in its entirety is a necessary underlying principle in this approach to help maintain minimum waste generation. Understanding consumer attitudes to these products, in particular when used in food or beverage systems, is critical in optimizing commercialization strategies.
This...
http://hdl.handle.net/11019/1135
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Alternative uses for co-products: harnessing the potential of valuable compounds from meat processing chains
(2018)
Mullen, Anne Maria; Álvarez, Carlos; Zeugolis, Dimitrios I.; Henchion, Maeve; O'Ne...
Alternative uses for co-products: harnessing the potential of valuable compounds from meat processing chains
(2018)
Mullen, Anne Maria; Álvarez, Carlos; Zeugolis, Dimitrios I.; Henchion, Maeve; O'Neill, Eileen; Drummond, Liana
Abstract:
Opportunities for exploiting the inherent value of protein-rich meat processing co-products, in the context of increased global demand for protein and for sustainable processing systems, are discussed. While direct consumption maybe the most profitable route for some, this approach is influenced greatly by local and cultural traditions. A more profitable and sustainable approach may be found in recognizing this readily available and under-utilised resource can provide high value components, such as proteins, with targeted high value functionality of relevance to a variety of sectors. Applications in food & beverages, petfood biomedical and nutrition arenas are discussed. Utilization of the raw material in its entirety is a necessary underlying principle in this approach to help maintain minimum waste generation. Understanding consumer attitudes to these products, in particular when used in food or beverage systems, is critical in optimizing commercialization strategies.
http://hdl.handle.net/10379/13009
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Analysis of gene expression in the bovine corpus luteum through generation and characterisation of 960 ests
(2018)
Casey, Orla M.; Fitzpatrick, Richard; McInerney, James O.; Morris, Dermot G.; Powell, R...
Analysis of gene expression in the bovine corpus luteum through generation and characterisation of 960 ests
(2018)
Casey, Orla M.; Fitzpatrick, Richard; McInerney, James O.; Morris, Dermot G.; Powell, Richard; Sreenan, Joseph M.
http://hdl.handle.net/10379/8951
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Analysis of stress-induced hepatic gene expression in rainbow trout (Oncorhynchus mykiss) selected for high- and low-responsiveness to stress
(2016)
Pemmasani, Jhansi K.; Pottinger, Tom G.; Cairns, Michael T.
Analysis of stress-induced hepatic gene expression in rainbow trout (Oncorhynchus mykiss) selected for high- and low-responsiveness to stress
(2016)
Pemmasani, Jhansi K.; Pottinger, Tom G.; Cairns, Michael T.
Abstract:
The production and welfare of intensively reared fish would be improved by reducing stress responsiveness. One approach to achieving this goal is selective breeding utilising stress-responsive genes as direct genetic markers of the desirable trait. As a first step in this process, microarray analysis has been carried out on liver tissues of rainbow trout selectively bred for high (HR) or low (LR) responsiveness to a stressor. Microarray hybridizations provided gene expression profiles for pooled samples of fish confined for 6 h, 24 h and 168 h and for individual fish (168 h only). 161 genes were shown to be differentially regulated in HR and LR fish during confinement exposure and eight of these gene expression profiles were validated by quantitative PCR. Genes of particular interest included intelectin-2 precursor which showed greater than 100-fold higher expression in HR fish compared to LR fish irrespective of whether the fish were confined or not; interferon inducible transmembr...
http://hdl.handle.net/10379/5833
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Anisotropy resolved multidimensional emission spectroscopy (ARMES) for the analysis and resolution of insulin oligomer emission
(2019)
Casamayou-Boucau, Yannick
Anisotropy resolved multidimensional emission spectroscopy (ARMES) for the analysis and resolution of insulin oligomer emission
(2019)
Casamayou-Boucau, Yannick
Abstract:
Protein aggregation is one of the biggest challenges affecting the manufacture and safe use of biopharmaceuticals. Despite efforts made to enhance protein stability [1], aggregation still remains a critical factor in product safety [2] as it can affect bioavailability and increase immunogenicity problems [3-4]. Aggregation can occur at every step of the bio-pharma process, from production to protein delivery and can be induced by a multitude of factors [5]. Detecting and quantifying protein aggregation is a considerable analytical challenge, due to the often unknown nature of the aggregates, their wide size range (nm to mm) [6] and the varying concentration ranges encountered. Within the aggregate family, being able to detect the soluble nm size aggregates is probably the most difficult/critical part as they form the seeds of bigger aggregation issues [7-8]. Many current methods for aggregation monitoring are either destructive, only capable of measuring large precipitated aggrega...
http://hdl.handle.net/10379/14931
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Anisotropy resolved multidimensional emission spectroscopy (ARMES): A new tool for protein analysis
(2016)
Groza, Radu Constantin; Ryder, Alan G.; Li, Boyan
Anisotropy resolved multidimensional emission spectroscopy (ARMES): A new tool for protein analysis
(2016)
Groza, Radu Constantin; Ryder, Alan G.; Li, Boyan
Abstract:
Structural analysis of proteins using the emission of intrinsic fluorophores is complicated by spectral overlap. Anisotropy resolved multidimensional emission spectroscopy (ARMES) overcame the overlap problem by the use of anisotropy, with chemometric analysis, to better resolve emission from different fluorophores. Total synchronous fluorescence scan (TSFS) provided information about all the fluorophores that contributed to emission while anisotropy provided information about the environment of each fluorophore. Here the utility of ARMES was demonstrated via study of the chemical and thermal denaturation of human serum albumin (HSA).Multivariate curve resolution (MCR) analysis of the constituent polarized emission ARMES data resolved contributions from four emitters: fluorescence from tryptophan (Trp), solvent exposed tyrosine (Tyr), Tyr in a hydrophobic environment, and room temperature phosphorescence (RTP) from Trp. The MCR scores, anisotropy, and literature validated these assi...
http://hdl.handle.net/10379/5507
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Antinociceptive effects of the GPR55 Antagonist CID16020046 injected into the rat anterior cingulate cortex
(2020)
Okine, Bright N.; Mc Laughlin, Gemma; Gaspar, Jessica C.; Harhen, Brendan; Roche, Miche...
Antinociceptive effects of the GPR55 Antagonist CID16020046 injected into the rat anterior cingulate cortex
(2020)
Okine, Bright N.; Mc Laughlin, Gemma; Gaspar, Jessica C.; Harhen, Brendan; Roche, Michelle; Finn, David P.
Abstract:
The G-protein coupled receptor, GPR55, modulates nociceptive processing. Given the expression of GPR55 in the anterior cingulate cortex (ACC), a key brain region involved in the cognitive and affective dimensions of pain, the present study tested the hypothesis that GPR55 signalling in the ACC facilitates inflammatory pain behaviour in rats. The expression of GPR55 in the ACC was confirmed by both western blotting and immunostaining, with evidence for neuronal localisation. Microinjection of the selective GPR55 antagonist CID16020046 into the ACC of adult male Sprague-Dawley rats significantly reduced second phase formalin-evoked nociceptive behaviour compared with vehicle-treated controls. CID16020046 administration was associated with a reduction in phosphorylation of extracellular signal-regulated kinase (ERK), a downstream target of GPR55 activation, in the ACC. Intra-ACC administration of CID16020046 prevented the formalin-induced increases in expression of mRNA coding for the ...
http://hdl.handle.net/10379/16254
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Association analysis identifies 65 new breast cancer risk loci
(2018)
Michailidou, Kyriaki; Lindström, Sara; Dennis, Joe; Beesley, Jonathan; Hui, Shirley; Ka...
Association analysis identifies 65 new breast cancer risk loci
(2018)
Michailidou, Kyriaki; Lindström, Sara; Dennis, Joe; Beesley, Jonathan; Hui, Shirley; Kar, Siddhartha; Lemaçon, Audrey; Soucy, Penny; Glubb, Dylan; Rostamianfar, Asha; Bolla, Manjeet K.; Wang, Qin; Tyrer, Jonathan; Dicks, Ed; Lee, Andrew; Wang, Zhaoming; Allen, Jamie; Keeman, Renske; Eilber, Ursula; French, Juliet D.
Abstract:
Breast cancer risk is influenced by rare coding variants in susceptibility genes, such as BRCA1, and many common, mostly non-coding variants. However, much of the genetic contribution to breast cancer risk remains unknown. Here we report the results of a genome-wide association study of breast cancer in 122,977 cases and 105,974 controls of European ancestry and 14,068 cases and 13,104 controls of East Asian ancestry(1). We identified 65 new loci that are associated with overall breast cancer risk at P &lt; 5 x 10(-8). The majority of credible risk single-nucleotide polymorphisms in these loci fall in distal regulatory elements, and by integrating in silico data to predict target genes in breast cells at each locus, we demonstrate a strong overlap between candidate target genes and somatic driver genes in breast tumours. We also find that heritability of breast cancer due to all single-nucleotide polymorphisms in regulatory features was 2-5-fold enriched relative to the geno...
http://hdl.handle.net/10379/12886
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Atp synthase f1 subunits recruited to centromeres by cenp-a are required for male meiosis
(2018)
Collins, Caitríona M.; Malacrida, Beatrice; Burke, Colin; Kiely, Patrick A.; Dunleavy, ...
Atp synthase f1 subunits recruited to centromeres by cenp-a are required for male meiosis
(2018)
Collins, Caitríona M.; Malacrida, Beatrice; Burke, Colin; Kiely, Patrick A.; Dunleavy, Elaine M.
Abstract:
The histone H3 variant CENP-A epigenetically defines the centromere and is critical for chromosome segregation. Here we report an interaction between CENP-A and subunits of the mitochondrial ATP synthase complex in the germline of male Drosophila. Furthermore, we report that knockdown of CENP-A, as well as subunits ATPsyn-alpha, -beta like (a testis-specific paralogue of ATPsyn-beta) and -gamma disrupts sister centromere cohesion in meiotic prophase I. We find that this disruption is likely independent of reduced ATP levels. We identify that ATPsyn-alpha and -beta like localise to meiotic centromeres and that this localisation is dependent on the presence of CENP-A. We show that ATPsyn-alpha directly interacts with the N-terminus of CENP-A in vitro and that truncation of its N terminus perturbs sister centromere cohesion in prophase I. We propose that the CENP-A N-terminus recruits ATPsyn-alpha and -beta like to centromeres to promote sister centromere cohesion in a nuclear function...
http://hdl.handle.net/10379/10854
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Atr-atrip kinase complex triggers activation of the fanconi anemia dna repair pathway
(2018)
Shigechi, T.; Tomida, J.; Sato, K.; Kobayashi, M.; Eykelenboom, J. K.; Pessina, F.; Zha...
Atr-atrip kinase complex triggers activation of the fanconi anemia dna repair pathway
(2018)
Shigechi, T.; Tomida, J.; Sato, K.; Kobayashi, M.; Eykelenboom, J. K.; Pessina, F.; Zhang, Y.; Uchida, E.; Ishiai, M.; Lowndes, N. F.; Yamamoto, K.; Kurumizaka, H.; Maehara, Y.; Takata, M.
http://hdl.handle.net/10379/13899
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Bacterial microcompartment-directed polyphosphate kinase promotes stable polyphosphate accumulation in E. coli
(2017)
Liang, Mingzhi; Frank, Stefanie; Lünsdorf, Heinrich; Warren, Martin J.; Prentice, Micha...
Bacterial microcompartment-directed polyphosphate kinase promotes stable polyphosphate accumulation in E. coli
(2017)
Liang, Mingzhi; Frank, Stefanie; Lünsdorf, Heinrich; Warren, Martin J.; Prentice, Michael B.
Abstract:
Processes for the biological removal of phosphate from wastewater rely on temporary manipulation of bacterial polyphosphate levels by phased environmental stimuli. In E. coli polyphosphate levels are controlled via the polyphosphate-synthesizing enzyme polyphosphate kinase (PPK1) and exopolyphosphatases (PPX and GPPA), and are temporarily enhanced by PPK1 overexpression and reduced by PPX overexpression. We hypothesised that partitioning PPK1 from cytoplasmic exopolyphosphatases would increase and stabilise E. coli polyphosphate levels. Partitioning was achieved by co-expression of E. coli PPK1 fused with a microcompartment-targeting sequence and an artificial operon of Citrobacter freundii bacterial microcompartment genes. Encapsulation of targeted PPK1 resulted in persistent phosphate uptake and stably increased cellular polyphosphate levels throughout cell growth and into the stationary phase, while PPK1 overexpression alone produced temporary polyphosphate increase and phosphate...
http://hdl.handle.net/10468/4211
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Beneficial modulation of the gut microbiota
(2020)
Cotter, Paul D.
Beneficial modulation of the gut microbiota
(2020)
Cotter, Paul D.
Abstract:
Conference abstract. 17th International Congress on Infectious Diseases. Invited presentation
As the scientific community continues to develop an ever-greater understanding of the composition and function of the human gut microbiota, and the role of specific microbial populations in health and disease, attention has turned to the tools that are at our disposal with respect to altering these microbes in a beneficial way. The options available include the use of diet, probiotics/prebiotics, antimicrobials and, potentially, exercise. Here, our recent investigations of the relationship between protein, bacteriocin producing probiotics and exercise and the gut microbiota and, in turn, health will be described.
http://hdl.handle.net/11019/2271
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Bioelectrochemical Haber-Bosch process: An ammonia-producing H-2/N-2 fuel cell
(2019)
Milton, Ross D.; Cai, Rong; Abdellaoui, Sofiene; Leech, Dónal; De Lacey, Antonio L.; Pi...
Bioelectrochemical Haber-Bosch process: An ammonia-producing H-2/N-2 fuel cell
(2019)
Milton, Ross D.; Cai, Rong; Abdellaoui, Sofiene; Leech, Dónal; De Lacey, Antonio L.; Pita, Marcos; Minteer, Shelley D.
Abstract:
Nitrogenases are the only enzymes known to reduce molecular nitrogen (N-2) to ammonia (NH3). By using methyl viologen (N, N'-dimethyl-4,4'-bipyridinium) to shuttle electrons to nitrogenase, N-2 reduction to NH3 can be mediated at an electrode surface. The coupling of this nitrogenase cathode with a bioanode that utilizes the enzyme hydrogenase to oxidize molecular hydrogen (H-2) results in an enzymatic fuel cell (EFC) that is able to produce NH3 from H-2 and N-2 while simultaneously producing an electrical current. To demonstrate this, a charge of 60 mC was passed across H-2/ N-2 EFCs, which resulted in the formation of 286 nmol NH3 mg (-1) MoFe protein, corresponding to a Faradaic efficiency of 26.4%.
This work is supported by a Marie Curie‐Skłodowska Individual Fellowship (Global) under the European Commission's Horizon 2020 Framework (project 654836 “Bioelectroammonia”), the Army Research Office MURI, and Spanish MINECO/FEDER, EU project CTQ2015‐71290‐R. We tha...
http://hdl.handle.net/10379/14968
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Bioelectrochemical haber-bosch process: an ammonia-producing h2/n2fuel cell
(2018)
Milton, Ross D.; Cai, Rong; Abdellaoui, Sofiene; Leech, Dónal; De Lacey, Antonio L.; Pi...
Bioelectrochemical haber-bosch process: an ammonia-producing h2/n2fuel cell
(2018)
Milton, Ross D.; Cai, Rong; Abdellaoui, Sofiene; Leech, Dónal; De Lacey, Antonio L.; Pita, Marcos; Minteer, Shelley D.
Abstract:
Nitrogenases are the only enzymes known to reduce molecular nitrogen (N-2) to ammonia (NH3). By using methyl viologen (N, N'-dimethyl-4,4'-bipyridinium) to shuttle electrons to nitrogenase, N-2 reduction to NH3 can be mediated at an electrode surface. The coupling of this nitrogenase cathode with a bioanode that utilizes the enzyme hydrogenase to oxidize molecular hydrogen (H-2) results in an enzymatic fuel cell (EFC) that is able to produce NH3 from H-2 and N-2 while simultaneously producing an electrical current. To demonstrate this, a charge of 60 mC was passed across H-2/ N-2 EFCs, which resulted in the formation of 286 nmol NH3 mg (-1) MoFe protein, corresponding to a Faradaic efficiency of 26.4%.
http://hdl.handle.net/10379/12906
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Brca1 is required for maintenance of phospho-chk1 and g2/m arrest during dna cross-link repair in dt40 cells
(2018)
Draga, Margarethe; Madgett, Elizabeth B.; Vandenberg, Cassandra J.; du Plessis, David; ...
Brca1 is required for maintenance of phospho-chk1 and g2/m arrest during dna cross-link repair in dt40 cells
(2018)
Draga, Margarethe; Madgett, Elizabeth B.; Vandenberg, Cassandra J.; du Plessis, David; Kaufmann, Aisling; Werler, Petra; Chakraborty, Prasun; Lowndes, Noel F.; Hiom, Kevin
Abstract:
The Fanconi anemia DNA repair pathway is pivotal for the efficient repair of DNA interstrand cross-links. Here, we show that FA-defective (Fancc(-)) DT40 cells arrest in G(2) phase following cross-link damage and trigger apoptosis. Strikingly, cell death was reduced in Fancc(-) cells by additional deletion of the BRCA1 tumor suppressor, resulting in elevated clonogenic survival. Increased resistance to cross-link damage was not due to loss of toxic BRCA1-mediated homologous recombination but rather through the loss of a G(2) checkpoint. This proapoptotic role also required the BRCA1-A complex member ABRAXAS (FAM175A). Finally, we show that BRCA1 promotes G(2) arrest and cell death by prolonging phosphorylation of Chk1 on serine 345 after DNA damage to sustain arrest. Our data imply that DNA-induced cross-link death in cells defective in the FA pathway is dependent on the ability of BRCA1 to prolong cell cycle arrest in G(2) phase.
http://hdl.handle.net/10379/11253
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Budding yeast rad9 is an atp-dependent rad53 activating machine
(2018)
Gilbert, Christopher S; Green, Catherine M; Lowndes, Noel F
Budding yeast rad9 is an atp-dependent rad53 activating machine
(2018)
Gilbert, Christopher S; Green, Catherine M; Lowndes, Noel F
Abstract:
We find budding yeast Rad9 in two distinct, large, and soluble complexes in cell extracts. The larger (greater than or equal to 850 kDa) complex, found in nondamaged cells, contains hypophosphorylated Rad9, whereas the smaller (560 kDa) complex, which forms after DNA damage, contains hyperphosphorylated Rad9 and Rad53. This smaller Rad9 complex is capable of catalyzing phosphorylation and release of active Rad53 kinase, a process requiring the kinase activity of Rad53. However, Mec1 and Tel1 are no longer required once the 560 kDa complex has been formed. We propose a model whereby Mec1/Tel1-dependent hyperphosphorylation of Rad9 results in formation of the smaller Rad9 complex and recruitment of Rad53. This complex then catalyzes activation of Rad53 by acting as a scaffold that brings Rad53 molecules into close proximity, facilitating Rad53 in trans autophosphorylation and subsequent release of activated Rad53.
http://hdl.handle.net/10379/9215
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Cdc7-dependent and -independent phosphorylation of claspin in the induction of the dna replication checkpoint
(2018)
Rainey, Michael; Harhen, Brendan; Wang, Guan-Nan; Murphy, Paul; Santocanale, Corrado
Cdc7-dependent and -independent phosphorylation of claspin in the induction of the dna replication checkpoint
(2018)
Rainey, Michael; Harhen, Brendan; Wang, Guan-Nan; Murphy, Paul; Santocanale, Corrado
Abstract:
Claspin is a critical mediator protein in the DNA replication checkpoint, responsible for ATR-dependent activation of the effector kinase Chk1. Cdc7, an essential kinase required for the initiation of DNA replication, can also interact with and phosphorylate Claspin. In this study we use small-molecule inhibitors of Cdc7 kinase to further understand the relationship between Cdc7, Claspin and Chk1 activation. We demonstrate that inhibition of Cdc7 kinase delays HU-induced phosphorylation of Chk1 but does not affect the maintenance of the replication checkpoint once it is established. We find that while chromatin association of Claspin is not affected by Cdc7 inhibition, Claspin phosphorylation is attenuated following HU treatment, which may be responsible for the altered kinetics of HU-induced Chk1 phosphorylation. We demonstrate that Claspin is an in vitro substrate of Cdc7 kinase, and using mass-spectrometry, we identify multiple phosphorylation sites that help to define a Cdc7 pho...
http://hdl.handle.net/10379/13577
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