TRANSVERSA: coordinación de actividades transversales del módulo básico de los grados en Biología y Ciencias del Mar

Esta red de investigación en docencia universitaria ha centrado sus objetivos en la coordinación de las actividades transversales en el módulo básico de los Grados en Biología y en Ciencias del Mar. Además, entre los intereses principales de la red se ha fijado la evaluación y propuestas de mejora de la planificación de las actividades con carga no presencial. En las sucesivas reuniones se ha contado con la participación de los delegados y delegadas de los distintos grupos y sus contribuciones han permitido incorporar la opinión del alumnado. Así, se ha establecido un primer análisis de la problemática a abordar en los siguientes años para mejorar la coordinación transversal y vertical en ambos grados, así como también se ha buscado una solución a todos los inconvenientes que han surgido a lo largo del semestre. Se plantea a su vez la mejora en la distribución de la carga semanal para evitar que haya semanas con mucha carga de horas no presenciales. Finalmente, se ha coordinado las competencias transversales en las asignaturas que comparten estos objetivos.

INVES: Docencia en Iniciación a la Investigación en Biología

Un equipo multidisciplinar de profesores y profesoras que imparten docencia en la asignatura Iniciación a la Investigación en Biología, constituyen la Red Docente INVES con el fin de desarrollar una metodología propia de trabajo en equipo, en coordinación con el profesorado de la asignatura Estadística, con la que se comparten objetivos de aprendizaje comunes. Durante el desarrollo de la asignatura, el alumnado diseña y ejecuta un proyecto de investigación bibliométrico de temática biológica sobre un tema actual y de interés. Con ello se favorece la adquisición de competencias transversales del módulo básico del título de grado. La dinámica de trabajo en grupo culmina en la edición de unas Jornadas Científicas, donde los estudiantes exponen los trabajos realizados. Se han consensuado modificaciones en las metodologías y actividades de aprendizaje, mejorando en la eficiencia de la experiencia de enseñanza-aprendizaje.

Dual regulation of P-glycoprotein expression by Trichostatin A in cancer cell lines

Background. It has been reported that the histone deacetylase inhibitor (iHDAc) trichostatin A (TSA) induces an increase in MDR1 gene transcription (ABCB1). This result would compromise the use of iHDACs in combination with other cytotoxic agents that are substrates of P-glycoprotein (Pgp). It has also been reported the use of alternative promoters by the ABCB1 gene and the existence of a traslational control of Pgp protein. Finally, the ABCB1 gene is located in a genetic locus with the nested gene RUNDC3B in the complementary DNA strand, raising the possibility that RUNDC3B expression could interfere with ABCB1 alternative promoter regulation. Methods. A combination of RT-PCR, real time RT-PCR, Western blot and drug accumulation assays by flow cytometry have been used in this study. Results. The iHDACs-induced increase in MDR1 mRNA levels is not followed by a subsequent increase in Pgp protein levels or activity in several pancreatic and colon carcinoma cell lines, suggesting a traslational control of Pgp in these cell lines. In addition, the MDR1 mRNA produced in these cell lines is shorter in its 5' end that the Pgp mRNA produced in cell lines expressing Pgp protein. The different size of the Pgp mRNA is due to the use of alternative promoters. We also demonstrate that these promoters are differentially regulated by TSA. The translational blockade of Pgp mRNA in the pancreatic carcinoma cell lines could be related to alterations in the 5' end of the MDR1 mRNA in the Pgp protein expressing cell lines. In addition, we demonstrate that the ABCB1 nested gene RUNDC3B expression although upregulated by TSA is independent of the ABCB1 alternative promoter used. Conclusions. The results show that the increase in MDR1 mRNA expression after iHDACs treatment is clinically irrelevant since this mRNA does not render an active Pgp protein, at least in colon and pancreatic cancer cell lines. Furthermore, we have demonstrated that TSA in fact, differentially regulates both ABCB1 promoters, downregulating the upstream promoter that is responsible for active P-glycoprotein expression. These results suggest that iHDACs such as TSA may in fact potentiate the effects of antitumoral drugs that are substrates of Pgp. Finally, we have also demonstrate that TSA upregulates RUNDC3B mRNA independently of the ABCB1 promoter in use.

Cross-talk and regulatory interactions between the essential response regulator RpaB and cyanobacterial circadian clock output

The response regulator RpaB (regulator of phycobilisome associated B), part of an essential two-component system conserved in cyanobacteria that responds to multiple environmental signals, has recently been implicated in the control of cell dimensions and of circadian rhythms of gene expression in the model cyanobacterium Synechococcus elongatus PCC 7942. However, little is known of the molecular mechanisms that underlie RpaB functions. In this study we show that the regulation of phenotypes by RpaB is intimately connected with the activity of RpaA (regulator of phycobilisome associated A), the master regulator of circadian transcription patterns. RpaB affects RpaA activity both through control of gene expression, a function requiring an intact effector domain, and via altering RpaA phosphorylation, a function mediated through the N-terminal receiver domain of RpaB. Thus, both phosphorylation cross-talk and coregulation of target genes play a role in the genetic interactions between the RpaA and RpaB pathways. In addition, RpaB∼P levels appear critical for survival under light:dark cycles, conditions in which RpaB phosphorylation is environmentally driven independent of the circadian clock. We propose that the complex regulatory interactions between the essential and environmentally sensitive NblS-RpaB system and the SasA-RpaA clock output system integrate relevant extra- and intracellular signals to the circadian clock.

Detecting KaiC Phosphorylation Rhythms of the Cyanobacterial Circadian Oscillator In Vitro and In Vivo

The central oscillator of the cyanobacterial circadian clock is unique in the biochemical simplicity of its components and the robustness of the oscillation. The oscillator is composed of three cyanobacterial proteins: KaiA, KaiB, and KaiC. If very pure preparations of these three proteins are mixed in a test tube in the right proportions and with ATP and MgCl2, the phosphorylation states of KaiC will oscillate with a circadian period, and these states can be analyzed simply by SDS-PAGE. The purity of the proteins is critical for obtaining robust oscillation. Contaminating proteases will destroy oscillation by degradation of Kai proteins, and ATPases will attenuate robustness by consumption of ATP. Here, we provide a detailed protocol to obtain pure recombinant proteins from Escherichia coli to construct a robust cyanobacterial circadian oscillator in vitro. In addition, we present a protocol that facilitates analysis of phosphorylation states of KaiC and other phosphorylated proteins from in vivo samples.