Temporally distinct roles of ATM and ROS in genotoxic- stress-dependent induction and maintenance of cellular senescence

Cells exposed to genotoxic stress induce cellular senescence through a DNA damage response (DDR) pathway regulated by ATM kinase and reactive oxygen species (ROS). Here, we show that the regulatory roles for ATM kinase and ROS differ during induction and maintenance of cellular senescence. Cells treated with different genotoxic agents were analyzed using specific pathway markers and inhibitors to determine that ATM kinase activation is directly proportional to the dose of the genotoxic stress and that senescence initiation is not dependent on ROS or the p53 status of cells. Cells in which ROS was quenched still activated ATM and initiated the DDR when insulted, and progressed normally to senescence. By contrast, maintenance of a viable senescent state required the presence of ROS as well as activated ATM. Inhibition or removal of either of the components caused cell death in senescent cells, through a deregulated ATM-ROS axis. Overall, our work demonstrates existence of an intricate temporal hierarchy between genotoxic stress, DDR and ROS in cellular senescence. Our model reports the existence of different stages of cellular senescence with distinct regulatory networks.

Structural and functional analysis of two universal stress proteins YdaA and YnaF from Salmonella typhimurium: possible roles in microbial stress tolerance

In many organisms ``Universal Stress Proteins'' CUSPS) are induced in response to a variety of environmental stresses. Here we report the structures of two USPs, YnaF and YdaA from Salmonella typhimurium determined at 1.8 angstrom and 2.4 angstrom resolutions, respectively. YnaF consists of a single USP domain and forms a tetrameric organization stabilized by interactions mediated through chloride ions. YdaA is a larger protein consisting of two tandem USP domains. Two protomers of YdaA associate to form a structure similar to the YnaF tetramer. YdaA showed ATPase activity and an ATP binding motif G-2X-G-9X-G(S/T/N) was found in its C-terminal domain. The residues corresponding to this motif were not conserved in YnaF although YnaF could bind ATP. However, unlike YdaA, YnaF did not hydrolyse ATP in vitro. Disruption of interactions mediated through chloride ions by selected mutations converted YnaF into an ATPase. Residues that might be important for ATP hydrolysis could be identified by comparing the active sites of native and mutant structures. Only the C-terminal domain of YdaA appears to be involved in ATP hydrolysis. The structurally similar N-terminal domain was found to bind a zinc ion near the segment equivalent to the phosphate binding loop of the C-terminal domain. Mass spectrometric analysis showed that YdaA might bind a ligand of approximate molecular weight 800 daltons. Structural comparisons suggest that the ligand, probably related to an intermediate in lipid A biosynthesis, might bind at a site close to the zinc ion. Therefore, the N-terminal domain of YdaA binds zinc and might play a role in lipid metabolism. Thus, USPs appear to perform several distinct functions such as ATP hydrolysis, altering membrane properties and chloride sensing. (C) 2015 Elsevier Inc. All rights reserved.

Insights into the Functional Roles of N-Terminal and C-Terminal Domains of Helicobacter pylori DprA

DNA processing protein A (DprA) plays a crucial role in the process of natural transformation. This is accomplished through binding and subsequent protection of incoming foreign DNA during the process of internalization. DprA along with Single stranded DNA binding protein A (SsbA) acts as an accessory factor for RecA mediated DNA strand exchange. H. pylori DprA (HpDprA) is divided into an N-terminal domain and a C-terminal domain. In the present study, individual domains of HpDprA have been characterized for their ability to bind single stranded (ssDNA) and double stranded DNA (dsDNA). Oligomeric studies revealed that HpDprA possesses two sites for dimerization which enables HpDprA to form large and tightly packed complexes with ss and dsDNA. While the N-terminal domain was found to be sufficient for binding with ss or ds DNA, C-terminal domain has an important role in the assembly of poly-nucleoprotein complex. Using site directed mutagenesis approach, we show that a pocket comprising positively charged amino acids in the N-terminal domain has an important role in the binding of ss and dsDNA. Together, a functional cross talk between the two domains of HpDprA facilitating the binding and formation of higher order complex with DNA is discussed.

Functional roles of N-terminal and C-terminal domains in the overall activity of a novel single-stranded DNA binding protein of Deinococcus radiodurans

Single-stranded DNA binding protein (Ssb) of Deinococcus radiodurans comprises N- and C-terminal oligonucleotide/oligosaccharide binding (OB) folds connected by a beta hairpin connector. To assign functional roles to the individual OB folds, we generated three Ssb variants: Ssb(N) (N-terminal without connector), Ssb(NC) (N-terminal with connector) and Ssb(C) (C-terminal), each harboring one OB fold. Both Ssb(N) and Ssb(NC) displayed weak single-stranded DNA (ssDNA) binding activity, compared to the full-length Ssb (Ssb(FL)). The level of ssDNA binding activity displayed by SsbC was intermediate between Ssb(FL) and Ssb(N). Ssb(C) and Ssb(FL) predominantly existed as homo-dimers while Ssb(NC)/Ssb(N) formed different oligomeric forms. In vitro, Ssb(NC) or Ssb(N) formed a binary complex with Ssb(C) that displayed enhanced ssDNA binding activity. Unlike Ssb(FL), Ssb variants were able to differentially modulate topoisomerase-I activity, but failed to stimulate Deinococcal RecA-promoted DNA strand exchange. The results suggest that the C-terminal OB fold is primarily responsible for ssDNA binding. The N-terminal OB fold binds weakly to ssDNA but is involved in multimerization. (C) 2015 The Authors. Published by Elsevier B.V. on behalf of the Federation of European Biochemical Societies. This is an open access article under the CC BY-NC-ND license.

Roles for Transcription Factors Sp1, NF-kappa B, IRF3, and IRF7 in Expression of the Human IFNL4 Gene

The expression of a biologically active human IFN4 depends on the presence of a frameshift deletion polymorphism within the first exon of the interferon lambda 4 (IFNL4) gene. In this report, we use the lung carcinoma-derived cell line, A549, which is genetically viable to express a functional IFN4, to address transcriptional requirements of the IFNL4 gene. We show that the GC-rich DNA-binding transcription factor (TF) specificity protein 1 (Sp1) is recruited to the IFNL4 promoter and has a role in induction of gene expression upon stimulation with viral RNA mimic poly(I:C). By using RNAi and overexpression strategies, we also show key roles in IFNL4 gene expression for the virus-inducible TFs, nuclear factor kappa-light-chain-enhancer of activated B cells (NF-B), IFN regulatory factor 3 (IRF3), and IRF7. Interestingly, we also observe that overexpression of IFN4 influences IFNL4 promoter activity, which may further be dependent on the retinoic acid-inducible gene-I (RIG-I)-like receptor pathway. Together, our work for the first time reports on the functional characterization of the human IFNL4 promoter.

Transposon mutagenesis in a hyper-invasive clinical isolate of Campylobacter jejuni reveals a number of genes with potential roles in invasion.

Transposon mutagenesis has been applied to a hyper-invasive clinical isolate of Campylobacter jejuni, 01/51. A random transposon mutant library was screened in an in vitro assay of invasion and 26 mutants with a significant reduction in invasion were identified. Given that the invasion potential of C. jejuni is relatively poor compared to other enteric pathogens, the use of a hyper-invasive strain was advantageous as it greatly facilitated the identification of mutants with reduced invasion. The location of the transposon insertion in 23 of these mutants has been determined; all but three of the insertions are in genes also present in the genome-sequenced strain NCTC 11168. Eight of the mutants contain transposon insertions in one region of the genome (approximately 14 kb), which when compared with the genome of NCTC 11168 overlaps with one of the previously reported plasticity regions and is likely to be involved in genomic variation between strains. Further characterization of one of the mutants within this region has identified a gene that might be involved in adhesion to host cells.

La equidad de género en la toma de decisiones a nivel de la Unidad de Producción Agropecuaria y de la Comunidad de Nandarola. Municipio de Nandaime (2013-2014)

La presente investigación se realizó en la comunidad de Nandarola, municipio de Nandaime, Nicaragua. 2013-2014. Los objetivos que se persiguieron fueron: analizar los diferentes factores que inciden en la equidad de género a través de los roles, la poca participación de la mujer en las labores agrícolas y sociales, así como capacitaciones, asistencia técnica, y la distribución del trabajo que desempeñan los miembros de cada unidad de producción agropecuaria, y en la Comunidad. Analizar el nivel de participación e integración de la mujer en la agricultura y labores domésticas. Conocer el grado de participación en la toma de decisiones de los recursos físicos y los cambios que se han venido dando en las diferentes épocas en el trabajo agropecuario. La metodología que se utilizó para la investigación fue tipo cualitativo-cuantitativo. En los métodos cualitativos se realizaron 17 entrevistas de preguntas abiertas para analizar el comportamiento y actitudes de hombres y mujeres, el grado de participación de la mujer en los procesos productivos en la agricultura, también se aplicaron como método cualitativo el grupo focal para conocer las diferentes expresiones de inequidad que ha vivido la mujer en la década de los ´80 y en la actualidad. En el análisis cuantitativo se realizaron 103 encuestas. En los resultados más importantes se encontró que el 8.33 % de las mujeres fueron beneficiados por la reforma agraria en la distribución de tierra, y los hombres con un 91.67%. Los problemas que enfrenta la población femenina es la falta de participación, en la toma de decisiones y el control de los recursos en la parte agropecuaria la mujer está ocupando apenas el 20 % en cambio el hombre con un 60%, dado que la mujer se limita a las actividades agrícolas y a la inserción del mercado. En el área pecuaria (ganado menor) la mujer colabora con un 71.43% y el hombre apenas aporta el 14.29%. En el aspecto organizativo las mujeres representan solamente el 20% del total de los miembros de las cooperativas, porque no tienen el tiempo suficiente de participar y son sometidas al conyugue. Otro dato importante que encontramos en la UPA como en la jefatura en el hogar el hombre representa un 63% y la mujer representa el 37%. La triple jornada (trabajo productivo, doméstico y reproductivo) de la mujer es uno de los problemas que enfrenta para participar de manera más sistemática en los diferentes ámbitos de su vida. En conclusión la mujer tiene menos acceso a los recursos físicos, por la falta de participación en las actividades agrícolas, también existe un dominio del hombre en todas las tomas de decisiones, tanto en el trabajo agropecuario como en el uso de los recursos de la unidad de producción agropecuaria, comunitario, y doméstico. Así mismo la distribución del trabajo familiar, lo doméstico está destinado para las hijas mujeres desde la temprana edad, y los hijos varones a las labores agrícolas y obreras.

How can IT professionals make the step into leadership roles?

Last year, Jisc began work with EDUCAUSE - the US organisation for IT professionals in higher education - to find out the skillset of the CIO of the future. One of the findings of our project was that many aspiring technology leaders find it difficult to make the step up. Louisa Dale, director Jisc group sector intelligence, talks through the learnings and opens a call for IT professionals to get involved in the next phase of work.

Relaciones amorosas de pareja en las trayectorias vitales de las mujeres encarceladas

[ES]La tesis doctoral analiza las experiencias amorosas de pareja de mujeres encarceladas, con el doble objetivo de visibilizar a las mujeres presas en el ámbito de las ciencias sociales y de introducir las especificidades de las mujeres encarceladas en los debates sociológicos y feministas acerca del amor. Las escasas aproximaciones al amor entre las mujeres presas han tendido a explicar sus relaciones de pareja desde el concepto de “dependencia emocional”, que, como se muestra en la tesis, presenta dos debilidades básicas, de un lado la tendencia a la psicologización y patologización de cuestiones de claro sustrato social; de otro la homogeneización de experiencias que presentan gran diversidad. Otra debilidad de ciertos análisis sobre las mujeres presas y aquellas excluidas socialmente, es que se han basado en concepciones sexistas acerca de las mujeres transgresoras como “malas mujeres”, por considerar que no cumplen con las expectativas culturales y sociales asociadas a los supuestos atributos de género. Esta tesis doctoral adopta una epistemología basada en la crítica feminista que busca modelos analíticos alejados de los estereotipos y la estigmatización de las mujeres transgresoras. Desde una perspectiva metodológica cualitativa, el trabajo de campo fue desarrollado en la cárcel de Nanclares de Oca (País Vasco) durante el 2008. Desarrollé un trabajo etnográfico de observación participante y entrevistas en profundidad semiestructuradas que elaboraban información sobre aspectos relativos a sus trayectorias de vida (familia de origen, vivienda, empleo, nivel educativo, situación penal y penitenciaria, estado de salud, etc.) y sus experiencias amorosas de pareja. En mi análisis, he rastreado la diversidad y variabilidad de las experiencias amorosas de las mujeres presas, el impacto del encarcelamiento en sus trayectorias amorosas y en la configuración de su intimidad, los elementos que hacen para estas mujeres del amor un “cautiverio”, y al mismo tiempo, las estrategias “liberadoras” que despliegan en sus desarrollos afectivos. El amor puede constituir un cautiverio para las mujeres ya que favorece la acomodación a unos roles de género que definen a las mujeres como dependientes y carentes de libertad. Al mismo tiempo, el amor se puede entender como una “estrategia emocional”, una forma de superar las consecuencias del encierro y de lograr ciertos estándares de “normalización” social, en un contexto en que se encuentran excluidas socialmente y fuertemente estigmatizadas.

Helena de Eurípides y los roles de género

Le Rane : Collana di Studi e Testi. Studi ; 55 - El teatro clásico en el marco de la cultura griega y su pervivencia en la cultura occidental ; 13

WIF India Workshop Report: Enhancing Women’s Roles in Fisheries in India, 1-3 February 2010, YUVA Centre, Navi Mumbai, India: report

The International Collective in Support of Fishworkers (ICSF) organized a workshop titled “Enhancing Women’s Roles in Fisheries in India” during 1-3 February 2010 in Navi Mumbai, India. The workshop was meant to discuss and analyze the role of women in fisheries, and reflect on issues facing women in fishing communities and organizations in contemporary India. Not only did the workshop expose the participants to women’s issues, organizational strategies, policies, schemes and legislation relevant to fishing communities, but it also stimulated discussions and exchange of views, which helped them explore a vision of a fisheries that sustains both communities and livelihoods. The workshop strengthened the linkages between diverse groups, and helped them search for strategies to protect the spaces of women in fisheries and their sources of livelihood. (PDF contains 61 pages)

Some ecological roles of Hippopotamus (Hippopotamus amphibious Linn. 1758) in fish production: possibilities for integrated fish-cum Agric production system

The common hippopotamus (Hippopotamus amphibious Linn. 1758) contributes to the productivity of aquatic systems where it lives. This paper reviews ecological roles of the hippo in this regard. Desk review of available literature information complemented with field observations were employed in the data collection. The ecological roles of the common hippopotamus being presented draw examples from East, West, Central and South African sub regions. The nutritional importance of the amphibious hippopotamus to rural communities was highlighted. In Southern Ethiopia, the Bodi, Bacha and Mura tribes eat hippo meat and this has led to severe hunting consequences on the wild populations of the animal. The important relationships between the hippopotamus and fish were presented. Hippopotamuses usually defecate in water and their excrements enrich the nutrients in the water resulting in favourable conditions for large fish populations. Some fish, including Labeo spp. were observed to feed on the micro-organisms and algae that grow on the skin of the hippotamus. A strong case was made for hippo-cum-fish integrated farm development in Nigeria based on ecological relationships so observed between the amphibious mammals and fish. This is one of the meeting points of fisheries and wildlife management that should be exploited for the benefits of the teeming Nigerian population

Novel roles of the SRP RNA in co-translational protein targeting

The signal recognition particle (SRP) and its receptor (SR) are universally conserved protein machineries that deliver nascent peptides to their proper destination. The SRP RNA is a universally conserved and essential component of SRP, which serves as the “catalyst” of the protein targeting cycle. The SRP RNA accelerates SRP-SR complex formation at the beginning of the protein targeting reaction, and triggers GTP hydrolysis and SRP-SR complex disassembly at the end. Here we combined biochemical and biophysical approaches to investigate the molecular mechanism of the functions of the SRP RNA. We found that two functional ends in the SRP RNA mediate distinct functions. The tetraloop end facilitates initial assembly of SRP and SR by mediating an electrostatic interaction with the Lys399 receptor, which ensures efficient and accurate substrate targeting. At the later stage of the SRP cycle, the SRP-SR complex relocalizes ~ 100 Angstrom to the 5’,3’-distal end of the RNA, a conformation crucial for GTPase activation and cargo handover. These results, combined with recent structural work, elucidate the functions of the SRP RNA during the protein targeting reaction.

Expression of eph-family receptor tyrosine kinases and ephrins in the tadpole of the frog Xenopus laevis, and possible roles in the development of retinotectal topography

Assembling a nervous system requires exquisite specificity in the construction of neuronal connectivity. One method by which such specificity is implemented is the presence of chemical cues within the tissues, differentiating one region from another, and the presence of receptors for those cues on the surface of neurons and their axons that are navigating within this cellular environment.

Connections from one part of the nervous system to another often take the form of a topographic mapping. One widely studied model system that involves such a mapping is the vertebrate retinotectal projection-the set of connections between the eye and the optic tectum of the midbrain, which is the primary visual center in non-mammals and is homologous to the superior colliculus in mammals. In this projection the two-dimensional surface of the retina is mapped smoothly onto the two-dimensional surface of the tectum, such that light from neighboring points in visual space excites neighboring cells in the brain. This mapping is implemented at least in part via differential chemical cues in different regions of the tectum.

The Eph family of receptor tyrosine kinases and their cell-surface ligands, the ephrins, have been implicated in a wide variety of processes, generally involving cellular movement in response to extracellular cues. In particular, they possess expression patterns-i.e., complementary gradients of receptor in retina and ligand in tectum- and in vitro and in vivo activities and phenotypes-i.e., repulsive guidance of axons and defective mapping in mutants, respectively-consistent with the long-sought retinotectal chemical mapping cues.

The tadpole of Xenopus laevis, the South African clawed frog, is advantageous for in vivo retinotectal studies because of its transparency and manipulability. However, neither the expression patterns nor the retinotectal roles of these proteins have been well characterized in this system. We report here comprehensive descriptions in swimming stage tadpoles of the messenger RNA expression patterns of eleven known Xenopus Eph and ephrin genes, including xephrin-A3, which is novel, and xEphB2, whose expression pattern has not previously been published in detail. We also report the results of in vivo protein injection perturbation studies on Xenopus retinotectal topography, which were negative, and of in vitro axonal guidance assays, which suggest a previously unrecognized attractive activity of ephrins at low concentrations on retinal ganglion cell axons. This raises the possibility that these axons find their correct targets in part by seeking out a preferred concentration of ligands appropriate to their individual receptor expression levels, rather than by being repelled to greater or lesser degrees by the ephrins but attracted by some as-yet-unknown cue(s).