Birth of a poly(A) tail: mechanisms and control of mRNA polyadenylation

During their synthesis in the cell nucleus, most eukaryotic mRNAs undergo a two-step 3′-end processing reaction in which the pre-mRNA is cleaved and released from the transcribing RNA polymerase II and a polyadenosine (poly(A)) tail is added to the newly formed 3′-end. These biochemical reactions might appear simple at first sight (endonucleolytic RNA cleavage and synthesis of a homopolymeric tail), but their catalysis requires a multi-faceted enzymatic machinery, the cleavage and polyadenylation complex (CPAC), which is composed of more than 20 individual protein subunits. The activity of CPAC is further orchestrated by Poly(A) Binding Proteins (PABPs), which decorate the poly(A) tail during its synthesis and guide the mRNA through subsequent gene expression steps. Here, we review the structure, molecular mechanism, and regulation of eukaryotic mRNA 3′-end processing machineries with a focus on the polyadenylation step. We concentrate on the CPAC and PABPs from mammals and the budding yeast, Saccharomyces cerevisiae, because these systems are the best-characterized at present. Comparison of their functions provides valuable insights into the principles of mRNA 3′-end processing. </p

Study based on electronic descriptors of the diastereoselective aza-diels-alder cycloaddition of [(1r)-10-(n,n-diethylsulfamoyl)isobornyl] 2h-azirine-3-carboxylate to e,e-1,4-diacetoxy-1,3-butadiene

Cycloaddition of chiral [(1R)-10-(N,N-diethylsulfamoyl)isobornyl] 2H-azirine-3-carboxylate to E,E-1,4-diacetoxy-1,3-butadiene shows complete diatereoselectivity giving a single cycloadduct (-)-(2S,5R,6R)-6-[(1R)-10-(N,N-diethylsulfamoyl)isobornyloxycarbonyl]-1-azabicyclo[4.1.0]hept-3-ene-2,5-diyldiacetate. Our main objective is to identify electronic/steric parameters capable of describing the observed tendencies of this reaction. The results of the calculations conclude that: even though the steric factors can play an important role at the initial steps of the reaction, at the transition states the behavior of several electronic parameters; like hardness, polarizability, aromaticity, charge transfer, etc is decisive enough to justify the obtained product. Finally, this work summarizes an exhaustive analysis of electronic descriptors and empirical reactivity principles, reaching a definitive and comprehensive explanation to the observed experimental result.Thanks to Fundação para a Ciência e Tecnologia (FCT) for partial financial support (project PTDC/QUI/67407/2006

Comparison of fungal spores concentrations measured with wideband integrated bioaerosol sensor and Hirst methodology

The aim of this work was to provide both a comparison of traditional and novel methodologies for airborne spores detection (i.e. the Hirst Burkard trap and WIBS-4) and the first quantitative study of airborne fungal concentrations in Payerne (Western Switzerland) as well as their relation to meteorological parameters. From the traditional method -Hirst trap and microscope analysis-, sixty-three propagule types (spores, sporangia and hyphae) were identified and the average spore concentrations measured over the full period amounted to 4145 ± 263.0 spores/m3. Maximum values were reached on July 19th and on August 6th. Twenty-six spore types reached average levels above 10 spores/m3. Airborne fungal propagules in Payerne showed a clear seasonal pattern, increasing from low values in early spring to maxima in summer. Daily average concentrations above 5000 spores/m3 were almost constant in summer from mid-June onwards. Weather parameters showed a relevant role for determining the observed spore concentrations. Coniferous forest, dominant in the surroundings, may be a relevant source for airborne fungal propagules as their distribution and predominant wind directions are consistent with the origin. The comparison between the two methodologies used in this campaign showed remarkably consistent patterns throughout the campaign. A correlation coefficient of 0.9 (CI 0.76–0.96) was seen between the two over the time period for daily resolutions (Hirst trap and WIBS-4). This apparent co-linearity was seen to fall away once increased resolution was employed. However at higher resolutions upon removal of Cladosporium species from the total fungal concentrations (Hirst trap), an increased correlation coefficient was again noted between the two instruments (R = 0.81 with confidence intervals of 0.74 and 0.86)

Emerging Views on the CTD Code

The C-terminal domain (CTD) of RNA polymerase II (Pol II) consists of conserved heptapeptide repeats that function as a binding platform for different protein complexes involved in transcription, RNA processing, export, and chromatin remodeling. The CTD repeats are subject to sequential waves of posttranslational modifications during specific stages of the transcription cycle. These patterned modifications have led to the postulation of the “CTD code” hypothesis, where stage-specific patterns define a spatiotemporal code that is recognized by the appropriate interacting partners. Here, we highlight the role of CTD modifications in directing transcription initiation, elongation, and termination. We examine the major readers, writers, and erasers of the CTD code and examine the relevance of describing patterns of posttranslational modifications as a “code.” Finally, we discuss major questions regarding the function of the newly discovered CTD modifications and the fundamental insights into transcription regulation that will necessarily emerge upon addressing those challenges

Mecanismos inductores y vías de la apoptosis en la diarrea vírica bovina

La apoptosis es una forma de muerte celular inducida por una gran variedad de estímulos, considerada como el resultado final de una cascada en la que intervienen una serie de enzimas denominadas caspasas, inducidas tanto por estímulos externos como internos. Entre éstas destacan las denominadas caspasas iniciadoras (2, 8, 9 y 10) y las caspasas ejecutoras o efectoras (3, 6 y 7). Existen dos vías principales reguladoras de la apoptosis: la vía extrínseca, inducida y mediada por receptores, la cual se inicia cuando un ligando apropiado se une a receptores de muerte localizados en la superficie celular, de modo que estas uniones ligando-receptor hacen que los dominios de muerte celular intracelulares y su unión a ciertas proteínas del citosol activen a las caspasas iniciadoras de esta vía (pe: caspasa 8), y la vía intrínseca o mitocondrial, gobernada por proteínas de la familia del Bcl-2 que agrupa un grupo de moléculas involucradas en la permeabilidad de la mitocondria, pudiéndose distinguir entre moléculas anti-apoptóticas y pro-apoptóticas. El desequilibrio en la expresión de estas moléculas incrementa la permeabilidad de la membrana mitocondrial, favoreciendo la liberación de proteínas al citosol y la activación de las caspasas iniciadora de esta vía (pe: caspasa 9). Una vez activadas las caspasas iniciadoras, éstas ejercerán su actividad proteolítica sobre las denominadas caspasas efectoras o ejecutoras (3, 6 y 7), las cuales, una vez activadas ejecutarán la muerte de forma irreversible. Junto a aspectos generales relacionados con los mecanismos y vías de la apoptosis, en este trabajo se abordan aspectos relacionados con dichos mecanismos en el transcurso de la diarrea vírica bovina, donde la apoptosis parece ser la responsable de la depleción linfoide y la leucopenia que caracterizan a esta enfermedad

Dynamics in Fip1 regulate eukaryotic mRNA 3^′ end processing

Cleavage and polyadenylation factor (CPF/CPSF) is a multiprotein complex essential for mRNA 3′ end processing in eukaryotes. It contains an endonuclease that cleaves pre-mRNAs, and a polymerase that adds a poly(A) tail onto the cleaved 3′ end. Several CPF subunits, including Fip1, contain intrinsically disordered regions (IDRs). IDRs within multiprotein complexes can be flexible, or can become ordered upon interaction with binding partners. Here, we show that yeast Fip1 anchors the poly(A) polymerase Pap1 onto CPF via an interaction with zinc finger 4 of another CPF subunit, Yth1. We also reconstitute a fully recombinant 850-kDa CPF. By incorporating selectively labeled Fip1 into recombinant CPF, we could study the dynamics of Fip1 within the megadalton complex using nuclear magnetic resonance (NMR) spectroscopy. This reveals that a Fip1 IDR that connects the Yth1- and Pap1-binding sites remains highly dynamic within CPF. Together, our data suggest that Fip1 dynamics within the 3′ end processing machinery are required to coordinate cleavage and polyadenylation.</p

El virus de la lengua azul como modelo para el estudio de los orbivirus

Al género Orbivirus pertenecen tres virus transmitidos a través de vector, que afectan al ganado domestico y a animales de vida libre (Lengua Azul, Peste Equina Africana y Enfermedad Hemorrágica Epizoótica de los Ciervos), y que han causado en las dos últimas décadas, especialmente el virus de la Lengua Azul (vLA), pérdidas económicas cuantiosas en prácticamente toda Europa y en los países del sur y el este del Mediterráneo. Es importante, por tanto, profundizar en el conocimiento sobre los mecanismos de actuación de estos virus, tomando como base al vLA considerado como el virus modelo para el estudio de este género, y en los mecanismos que modulan la respuesta inmunológica del hospedador para poder mejorar el control de la enfermedad mediante vacunas más eficaces. Hasta el momento se desconocen aspectos tan importantes como la duración de la viremia en animales infectados, el papel de la inmunidad celular desencadenada en el hospedador o los mecanismos de “hibernación” que permitan al vLA “reaparecer” tras un periodo de ausencia del vector. Además, el hecho de que los tres compartan los mismos vectores, hace necesario analizar el riesgo de entrada de los virus de la Peste Equina Africana y dela Enfermedad Hemorrágica Epizoótica de los Ciervos en España, -que actualmente se detectan en los países del África Subsahariana y en el sur y este del Mediterráneo, respectivamente-. Por último, se considera también imprescindible mejorar las técnicas de diagnóstico para el virus de la Enfermedad Hemorrágica Epizoótica de los Ciervos y el virus de la Peste Equina Africana y desarrollar nuevos ensayos basados en las cepas circulantes por el área mediterránea

Geology and paleontology of Tresjuncos (Cuenca, Spain), a new diatomaceous deposit with Konservat-Lagerstätte characteristics from the European late Miocene.

Purpose A new Iberian Konservat-Lagersta¨tte discovered in Tresjuncos (Cuenca, Spain) is described and interpreted based on geological and paleontological information. Methods The samples were studied using conventional mineralogical (powder X-ray diffraction), petrographic (optical and scanning electron microscopy) and geochemical (X-ray energy-dispersive spectrometry) techniques. Results Based upon geological context, stratigraphy, lithology, sedimentology and origin of the rocks, we describe and interpret the new fossil site as originated in a lacustrine environment. The micromammals found in the Tresjuncos 1 outcrop suggest that the main site is Turolian in age (uppermost Miocene), continental chronological unit MN13. The fossils are preserved in laminates comprising rhythmic, millimeterscale alternations of diatomites and calcareous diatomites. An analysis of the lake paleobiota includes amphibians, crustaceans, insects, plants, and diatoms. The diatoms are dominated by Cyclotella iris. The whole assemblage is indicative of a planktic community developed in somewhat alkaline waters. An extremely well preserved premetamorphic Pelobates (Pelobatidae) tadpole was discovered; it is remarkable because of its gigantic size. Its digestive tract predominantly contains the rare diatom Navicula halionata var directa, likely associated with the shore vegetation in which it presumably lived and fed. The arthropods form a typical lacustrine aquatic assemblage, lacking fish. This assemblage is made up of small filtering crustaceans and of both predator and nonpredator insects of diverse sizes. Conclusions The available evidence on Tresjuncos shows the extraordinary potential for paleontological information that can be expected from this new Konservat-Lagersta¨tte, which constitutes a significant finding that improves our understanding of continental late Miocene Mediterranean environments

Lengua azul: vacunas, inmunomoduladores e inmunidad protectora

La Lengua Azul, está producida por un virus ARN del género Orbivirus (familia Reoviridae), considerado como el virus prototipo de este género, del que se conocen al menos 24 serotipos diferentes, no todos patógenos, entre los que no existe inmunidad cruzada, lo que difi culta las estrategias de vacunación. En las dos últimas décadas, y más recientemente desde el verano de 2006, esta enfermedad ha provocado importantes pérdidas económicas, no sólo en las zonas de Europa periódicamente afectadas como los países de la cuenca Mediterránea, sino prácticamente en toda Europa. Los planes de vacunación puestos en marcha por las autoridades sanitarias, han revelado la existencia de reacciones adversas, así como la falta de protección de las vacunas en un elevado porcentaje de casos. En este trabajo pretendemos poner de manifi esto la importancia de conocer los mecanismos inmunológicos que se desarrollan tanto en animales infectados como en animales vacunados. Los mecanismos de acción de cada uno de estos serotipos varían completamente dependiendo de la especie y de la raza afectada. Hasta la fecha son escasos los trabajos in vivo que hayan centrado sus esfuerzos en una caracterización pormenorizada de los mecanismos patogénicos y de la repuesta inmune de cada uno de los serotipos patógenos en las principales especies afectadas por la enfermedad. Sólo con el conocimiento de los mecanismos de acción del virus y con el estudio de los mecanismos que controlar y modulan la respuesta inmune podremos desarrollar herramientas (nuevos adyuvantes, aplicación de inmunomoduladores, etc) que nos permitan mejorar la vacunas existentes, reduciendo las reacciones adversas que producen y potenciando su protección