ABI1 and PP2CA Phosphatases Are Negative Regulators of Snf1-Related Protein Kinase1 Signaling in Arabidopsis

Plant survival under environmental stress requires the integration of multiple signaling pathways into a coordinated response, but the molecular mechanisms underlying this integration are poorly understood. Stress-derived energy deprivation activates the Snf1-related protein kinases1 (SnRK1s), triggering a vast transcriptional and metabolic reprogramming that restores homeostasis and promotes tolerance to adverse conditions. Here, we show that two clade A type 2C protein phosphatases (PP2Cs), established repressors of the abscisic acid (ABA) hormonal pathway, interact with the SnRK1 catalytic subunit causing its dephosphorylation and inactivation. Accordingly, SnRK1 repression is abrogated in double and quadruple pp2c knockout mutants, provoking, similarly to SnRK1 overexpression, sugar hypersensitivity during early seedling development. Reporter gene assays and SnRK1 target gene expression analyses further demonstrate that PP2C inhibition by ABA results in SnRK1 activation, promoting SnRK1 signaling during stress and once the energy deficit subsides. Consistent with this, SnRK1 and ABA induce largely overlapping transcriptional responses. Hence, the PP2C hub allows the coordinated activation of ABA and energy signaling, strengthening the stress response through the cooperation of two key and complementary pathways.

Dissection of miRNA Pathways Using Arabidopsis Mesophyll Protoplasts

MicroRNAs (miRNAs) control gene expression mostly post-transcriptionally by guiding transcript cleavage and/or translational repression of complementary mRNA targets, thereby regulating developmental processes and stress responses. Despite the remarkable expansion of the field, the mechanisms underlying miRNA activity are not fully understood. In this article, we describe a transient expression system in Arabidopsis mesophyll protoplasts, which is highly amenable for the dissection of miRNA pathways. We show that by transiently overexpressing primary miRNAs and target mimics, we can manipulate miRNA levels and consequently impact on their targets. Furthermore, we developed a set of luciferase-based sensors for quantifying miRNA activity that respond specifically to both endogenous and overexpressed miRNAs and target mimics. We demonstrate that these miRNA sensors can be used to test the impact of putative components of the miRNA pathway on miRNA activity, as well as the impact of specific mutations, by either overexpression or the use of protoplasts from the corresponding mutants. We further show that our miRNA sensors can be used for investigating the effect of chemicals on miRNA activity. Our cell-based transient expression system is fast and easy to set up, and generates quantitative results, being a powerful tool for assaying miRNA activity in vivo.

Complex genetic findings in a female patient with pyruvate dehydrogenase complex deficiency: Null mutations in the PDHX gene associated with unusual expression of the testis-specific PDHA2 gene in her somatic cells

Human pyruvate dehydrogenase complex (PDC) catalyzes a key step in the generation of cellular energy and is composed by three catalytic elements (E1, E2, E3), one structural subunit (E3-binding protein), and specific regulatory elements, phosphatases and kinases (PDKs, PDPs). The E1α subunit exists as two isoforms encoded by different genes: PDHA1 located on Xp22.1 and expressed in somatic tissues, and the intronless PDHA2 located on chromosome 4 and only detected in human spermatocytes and spermatids. We report on a young adult female patient who has PDC deficiency associated with a compound heterozygosity in PDHX encoding the E3-binding protein. Additionally, in the patient and in all members of her immediate family, a full-length testis-specific PDHA2 mRNA and a 5′UTR-truncated PDHA1 mRNA were detected in circulating lymphocytes and cultured fibroblasts, being bothmRNAs translated into full-length PDHA2 and PDHA1 proteins, resulting in the co-existence of both PDHA isoforms in somatic cells.Moreover, we observed that DNA hypomethylation of a CpG island in the coding region of PDHA2 gene is associatedwith the somatic activation of this gene transcription in these individuals. This study represents the first natural model of the de-repression of the testis-specific PDHA2 gene in human somatic cells, and raises some questions related to the somatic activation of this gene as a potential therapeutic approach for most forms of PDC deficiency.

Violencia e invisibilidad indígena : La cuestión de los pueblos originarios durante el primer peronismo

El presente trabajo busca profundizar en el análisis de los vínculos existentes entre el primer gobierno peronista y la cuestión indígena, centrándose en las políticas gubernamentales desarrolladas por el gobierno de Perón frente a dos fuertes conflictos violentos que tuvieron lugar entre 1946 y 1947: el "Malón de la paz", la movilización indígena llevada a cabo entre mayo y agosto de 1947, cuando 174 kollas caminaron 2000 kilómetros desde la Puna y el valle de Orán hasta la Capital Federal para reclamar por la titularidad de sus tierras, en manos de terratenientes y en denuncia de las condiciones de explotación en las que trabajaban; y "Masacre de Rincón Bomba", el conflicto desarrollado en una pequeña localidad de Formosa, cuando indígenas de comunidades wichi, tobas y principalmente pilagás fueron masacradas por la Gendarmería Nacional en un confuso episodio, que sale a la luz hace pocos años. El objetivo en ambos puntos es doble: por un lado analizar la relación entre los intereses e intenciones del gobierno de Juan Domingo Perón para con las comunidades originarias, visibilizando su existencia y sus condiciones de vida en tanto sujetos de derechos históricamente vulnerados. Por otro, abrir el debate historiográfico sobre el quehacer de los historiadores respecto de una temática que ha sido silenciada durante décadas, negando la existencia y la identidad de los pueblos originarios

Politics and religion; a study in Scottish history from the reformation to the revolution,

I. The French alliance. Condition of the church. The eve of the reformation, 1550-1559. The war of reformation, 1559-1560. The reformation parliament. John Knox. Maitland and Mary Stewart, 1561-1567. Civil war, 1568-1573. The new religion. Church and state. Bishops and presbyters, 1572-1625. The reign of the moderates. The national covenant, 1625-1638. Presbytery restored, 1638. The Glasgow assembly.--II. The covenant in arms, 1639-1641. The solemn league and covenant, 1641-1643. The royalist reaction, 1644-1648. The theocratic experiment, 1648-1651. The reign of the zealots. The restoration, 1651-1663. The Pentland rising, 1663-1667. The Leighton group, 1667-1674. The Bothwell rising, 1674-1680. Fanaticism and repression, 1680-1685. The revolution, 1685-1688. The revolution settlement, 1688-1695.

Bulletin.

Vol. 1, no. 2 (May, 1918) with title, A handbook of common garden pests, has no General bulletin numbering.

Choix de discours et 'opinions /

About political matters, repression, censorship, the civil code, etc.

Étude sur les champignons parasites (parodiellinacées, inclus erysiphées)

At head of title: Ministère de l'agriculture. Direction des services sanitaires et scientifiques et de la repression des fraudes.

Violencia e invisibilidad indígena : La cuestión de los pueblos originarios durante el primer peronismo

El presente trabajo busca profundizar en el análisis de los vínculos existentes entre el primer gobierno peronista y la cuestión indígena, centrándose en las políticas gubernamentales desarrolladas por el gobierno de Perón frente a dos fuertes conflictos violentos que tuvieron lugar entre 1946 y 1947: el "Malón de la paz", la movilización indígena llevada a cabo entre mayo y agosto de 1947, cuando 174 kollas caminaron 2000 kilómetros desde la Puna y el valle de Orán hasta la Capital Federal para reclamar por la titularidad de sus tierras, en manos de terratenientes y en denuncia de las condiciones de explotación en las que trabajaban; y "Masacre de Rincón Bomba", el conflicto desarrollado en una pequeña localidad de Formosa, cuando indígenas de comunidades wichi, tobas y principalmente pilagás fueron masacradas por la Gendarmería Nacional en un confuso episodio, que sale a la luz hace pocos años. El objetivo en ambos puntos es doble: por un lado analizar la relación entre los intereses e intenciones del gobierno de Juan Domingo Perón para con las comunidades originarias, visibilizando su existencia y sus condiciones de vida en tanto sujetos de derechos históricamente vulnerados. Por otro, abrir el debate historiográfico sobre el quehacer de los historiadores respecto de una temática que ha sido silenciada durante décadas, negando la existencia y la identidad de los pueblos originarios

Speaking in Tongues: Translating Yiddish in the 20th Century, pt. 3

Writing in Tongues examines the complexities of translating Yiddish literature at a time when the Yiddish language is in decline. After the Holocaust, Soviet repression, and American assimilation, the survival of traditional Yiddish literature depends on translation, yet a few Yiddish classics have been translated repeatedly while many others have been ignored. Anita Norich traces historical and aesthetic shifts through versions of these canonical texts, and she argues that these works and their translations form an enlightening conversation about Jewish history and identity.

Helene & Writing Abuse, Shame, and Death: A Poetics of Spirit within the Failing Body

Thesis (Master's)--University of Washington, 2016-06

Investigating the Effect of Histone H4 Sumoylation on Chromatin Structure and Function

Thesis (Ph.D.)--University of Washington, 2016-06

Systematic characterization of the zinc-finger-containing proteins in the mouse transcriptome

Zinc-finger-containing proteins can be classified into evolutionary and functionally divergent protein families that share one or more domains in which a zinc ion is tetrahedrally coordinated by cysteines and histidines. The zinc finger domain defines one of the largest protein superfamilies in mammalian genomes; 46 different conserved zinc finger domains are listed in InterPro (http://www.ebi.ac.uk/InterPro). Zinc finger proteins can bind to DNA, RNA, other proteins, or lipids as a modular domain in combination with other conserved structures. Owing to this combinatorial diversity, different members of zinc finger superfamilies contribute to many distinct cellular processes, including transcriptional regulation, mRNA stability and processing, and protein turnover. Accordingly, mutations of zinc finger genes lead to aberrations in a broad spectrum of biological processes such as development, differentiation, apoptosis, and immunological responses. This study provides the first comprehensive classification of zinc finger proteins in a mammalian transcriptome. Specific detailed analysis of the SP/Kruppel-like factors and the E3 ubiquitin-ligase RING-H2 families illustrates the importance of such an analysis for a more comprehensive functional classification of large protein families. We describe the characterization of a new family of C2H2 zinc-finger-containing proteins and a new conserved domain characteristic of this family, the identification and characterization of Sp8, a new member of the Sp family of transcriptional regulators, and the identification of five new RING-H2 proteins.

Distinct cis-elements in the Asparagus officinalis asparagine synthetase promoter respond to carbohydrate and senescence signals

The Asparagus officinalis L. asparagine (Asn) synthetase (AS) promoter was analysed for elements responding to carbohydrate and senescence signals. Transgenic Arabidopsis thaliana L. plants containing deletion constructs of the –1958 bp AS promoter linked to the β-glucuronidase (GUS) reporter gene (AS::GUS) were analysed by measuring GUS specific activity. Inclusion of sucrose (Suc), glucose (Glc) or fructose (Fru) in plant media repressed levels of GUS activity in –1958AS::GUS plants, regardless of the light environment, with increases in GUS found 1 d after incubation on Suc-lacking media. Hexokinase is likely to be involved in the signal pathway, as Suc, Glc, Fru, 2-deoxy-d-glucose and mannose were more effective repressors than 3-O-methylglucose, and the hexokinase inhibitor mannoheptulose reduced repression. Plants containing AS::GUS constructs with deletions that reduced the promoter to less than –405 bp did not show low sugar induction. AS::GUS activity was significantly higher in excised leaves induced to senesce by dark storage for 24 h, compared to fresh leaves, for lines containing at least –640 bp of the AS promoter but not those with –523 bp or smaller promoter fragments. Fusion of the –640 to –523 bp region to a –381AS::GUS construct generated a promoter that retained senescence induction but lacked low sugar induction. Alignment of this region to the 33-bp senescence-related sequence of the Arabidopsis and Brassica napus L. SAG12 promoters identified the sequence TTGCACG as being conserved in all the promoters, and which may be an important senescence-responsive element.

Gene expression during early ascidian metamorphosis requires signaling by Hemps, an EGF-like protein

Hemps, a novel epidermal growth factor (EGF)-like protein, is expressed during larval development and early metamorphosis in the ascidian Herdmania curvata and plays a direct role in triggering metamorphosis. In order to identify downstream genes in the Hemps pathway we used a gene expression profiling approach, in which we compared post-larvae undergoing normal metamorphosis with larval metamorphosis blocked with an anti-Hemps antibody. Molecular profiling revealed that there are dynamic changes in gene expression within the first 30 minutes of normal metamorphosis with a significant portion of the genome (approximately 49%) being activated or repressed. A more detailed analysis of the expression of 15 of these differentially expressed genes through embryogenesis, larval development and metamorphosis revealed that while there is a diversity of temporal expression patterns, a number of genes are transiently expressed during larval development and metamorphosis. These and other differentially expressed genes were localised to a range of specific cell and tissue types in Herdmania larvae and post-larvae. The expression of approximately 24% of the genes that were differentially expressed during early metamorphosis was affected in larvae treated with the anti-Hemps antibody. Knockdown of Hemps activity affected the expression of a range of genes within 30 minutes of induction, suggesting that the Hemps pathway directly regulates early response genes at metamorphosis. In most cases, it appears that the Hemps pathway contributes to the modulation of gene expression, rather than initial gene activation or repression. A total of 151 genes that displayed the greatest alterations in expression in response to anti-Hemps antibody were sequenced. These genes were implicated in a range of developmental and physiological roles, including innate immunity, signal transduction and in the regulation of gene transcription. These results suggest that there is significant gene activity during the very early stages of H. curvata metamorphosis and that the Hemps pathway plays a key role in regulating the expression of many of these genes.