Person: Rodríguez Escudero, María Isabel
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First Name
María Isabel
Last Name
Rodríguez Escudero
Affiliation
Universidad Complutense de Madrid
Faculty / Institute
Farmacia
Department
Microbiología y Parasitología
Area
Microbiología
Identifiers
11 results
Search Results
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Item Heterologous mammalian Akt disrupts plasma membrane homeostasis by taking over TORC2 signaling in Saccharomyces cerevisiae(Scientific Reports, 2018) Rodríguez Escudero, María Isabel; Fernández-Acero Bascones, Teresa; Jiménez Cid, Víctor; Molina Martín, MaríaThe Akt protein kinase is the main transducer of phosphatidylinositol-3,4,5-trisphosphate (PtdIns3,4,5P3) signaling in higher eukaryotes, controlling cell growth, motility, proliferation and survival. By co-expression of mammalian class I phosphatidylinositol 3-kinase (PI3K) and Akt in the Saccharomyces cerevisiae heterologous model, we previously described an inhibitory effect on yeast growth that relied on Akt kinase activity. Here we report that PI3K-Akt expression in yeast triggers the formation of large plasma membrane (PM) invaginations that were marked by actin patches, enriched in PtdIns4,5P2 and associated to abnormal intracellular cell wall deposits. These effects of Akt were mimicked by overproduction of the PtdIns4,5P2 effector Slm1, an adaptor of the Ypk1 and Ypk2 kinases in the TORC2 pathway. Although Slm1 was phosphorylated in vivo by Akt, TORC2-dependent Ypk1 activation did not occur. However, PI3K-activated Akt suppressed the lethality derived from inactivation of either TORC2 or Ypk protein kinases. Thus, heterologous co-expression of PI3K and Akt in yeast short-circuits PtdIns4,5P2- and TORC2-signaling at the level of the Slm-Ypk complex, overriding some of its functions. Our results underscore the importance of phosphoinositide-dependent kinases as key actors in the homeostasis and dynamics of the PM.Item Functional analysis of PTEN variants of unknown significance from PHTS patients unveils complex patterns of PTEN biological activity in disease(Eur J Hum Genet., 2023) Torices, Leire; Mingo, Janire; Rodríguez Escudero, María Isabel; Fernández-Acero Bascones, Teresa; Luna, Sandra; Nunes-Xavier, Caroline E.; López, José I.; Mercadillo, Fátima; Currás, María; Urioste, Miguel; Molina Martín, María; Jiménez Cid, Víctor; Pulido, RafaelHeterozygous germline mutations in PTEN gene predispose to hamartomas and tumors in different tissues, as well as to neurodevelopmental disorders, and define at genetic level the PTEN Hamartoma Tumor Syndrome (PHTS). The major physiologic role of PTEN protein is the dephosphorylation of phosphatidylinositol (3,4,5)-trisphosphate (PIP3), counteracting the pro-oncogenic function of phosphatidylinositol 3-kinase (PI3K), and PTEN mutations in PHTS patients frequently abrogate PTEN PIP3 catalytic activity. PTEN also displays non-canonical PIP3-independent functions, but their involvement in PHTS pathogeny is less understood. We have previously identified and described, at clinical and genetic level, novel PTEN variants of unknown functional significance in PHTS patients. Here, we have performed an extensive functional characterization of these PTEN variants (c.77 C > T, p.(Thr26Ile), T26I; c.284 C > G, p.(Pro95Arg), P95R; c.529 T > A, p.(Tyr177Asn), Y177N; c.781 C > G, p.(Gln261Glu), Q261E; c.829 A > G, p.(Thr277Ala), T277A; and c.929 A > G, p.(Asp310Gly), D310G), including cell expression levels and protein stability, PIP3-phosphatase activity, and subcellular localization. In addition, caspase-3 cleavage analysis in cells has been assessed using a C2-domain caspase-3 cleavage-specific anti-PTEN antibody. We have found complex patterns of functional activity on PTEN variants, ranging from loss of PIP3-phosphatase activity, diminished protein expression and stability, and altered nuclear/cytoplasmic localization, to intact functional properties, when compared with PTEN wild type. Furthermore, we have found that PTEN cleavage at the C2-domain by the pro-apoptotic protease caspase-3 is diminished in specific PTEN PHTS variants. Our findings illustrate the multifaceted molecular features of pathogenic PTEN protein variants, which could account for the complexity in the genotype/phenotype manifestations of PHTS patients.Item MicroMundo@UCM: Ciencia ciudadana y aprendizaje-servicio frente a la resistencia a antibióticos desde la comunidad educativa(2023) Jiménez Cid, Víctor; Ayllón Santiago, Tania; Amaro Torres, Francisco; Calvo De Pablo, Pilar; Domenech Lucas, Miriam; De Francisco Martínez, Patricia; Patiño Álvarez, Aurora Belén; Vázquez Estévez, María Covadonga Inmaculada; Barbero Úriz, Óscar; Blesa Esteban, Alba Mercedes; Borrajo López, Ana; Cortés Prieto, Isabel; Fernández-Acero Bascones, Teresa; Fernández-Vega Granado, Alejandro; García Pastor, Lucía; González Rubio, Gema; Lavilla García, Beatriz; López Montesino, Sara; Martínez López, Raquel María; Molina Martín, María; Mascaraque Martín, Victoria; Parra Giraldo, Claudia Marcela; Rodríguez Escudero, María Isabel; Rodríguez Fernández, Carmina; Román González, Elvira; Rubio Lozano, Alba Victoria; Del Val Oriza, Elba; Valentí Sanguino, Marta; Bezos Garrido, Javier; Borrero Del Pino, Juan; Díaz Formoso, Lara; Escudero García-Calderón, José Antonio; García Benzaquén, Nerea; González Zorn, Bruno; Hipólito Carrillo de Albornoz, Alberto; Muñoz Atienza, Estefanía; Pérez Sancho, Marta; Pulido Vadillo, Mario; Romero Martínez, Beatriz; Sánchez Méndez, Irene; Serna Bernaldo, Carlos; Suárez Rodríguez, Mónica; Vergara González, Ester; Arias López, Patricia; González de Figueras, Carolina; Prieto Orzanco, Alicia; Sempere García, Julio; Gil Serna, Jessica; Maestro García-Donas, María Beatriz; García García, Aina; Gil Serna, JessicaSegún ha establecido la OMS y la Asamblea de las Naciones Unidas, la resistencia a los antibióticos es una de las prioridades en Salud Global para el s. XXI, al mismo nivel que la preparación frente a pandemias emergentes y el desarrollo de vacunas y tratamientos frente a tuberculosis, malaria, HIV y otras enfermedades infecciosas. El abordaje de estos problemas sólo será eficaz desde la perspectiva multidisciplinar One Health (Un Mundo: Una Salud), que integra Salud Medioambiental, Animal y Humana. Desde 2016 la UCM ha sido pionera en implicar a las comunidades universitaria y preuniversitaria en la estrategia de aprendizaje activo mediante ApS “MicroMundo” (www.ucm.es/small-world-initiative), un proyecto multidisciplinar basado en ApS en el ámbito de la Biomedicina. Se trata de una adaptación al ApS de la estrategia internacional de studentsourcing Tiny Earth (www.tinyearthnet.org). MicroMundo implica a estudiantes de ESO y Bachillerato a participar como investigadores en un proyecto real para el descubrimiento de nuevos antibióticos a partir de muestras de suelos. El objetivo final del servicio es acercar la cultura científica, la perspectiva One Health y la investigación biomédica a la sociedad. Para ello, colaboramos con una red de colegios e institutos de la Comunidad de Madrid. Un objetivo adicional es fomentar la vocación por Grados STEM y por la I+D. Para lograr estos objetivos, los estudiantes universitarios, objeto del aprendizaje, se responsabilizan de facilitar a los jóvenes estudiantes el entrenamiento y material necesarios para llevar a cabo los experimentos, así como dirigir el trabajo de investigación y diversas estrategias de divulgación hacia la comunidad. El éxito del proyecto en la UCM ha inspirado su implementación en más de 30 universidades en España y Portugal y ha merecido el Premio PRAN 2021 a la comunicación y sensibilización de la población sobre la resistencia a los antibióticos. En esta publicación presentamos la memoria final del proyecto MicroMundo@UCM realizado durante el curso 2022-23 en 33 centros educativos de la Comunidad de Madrid.- Project number: 265
Item SWI@UCM 2.0: Consolidación de Small World Initiative: descubrimiento y uso racional de antibióticos mediante aprendizaje-servicio en la Comunidad de Madrid(2018) Jiménez Cid, Víctor; Rodríguez Escudero, María Isabel; Díez Orejas, Rosalía María; Molina Martín, María; Rodríguez Fernández, Carmina; Navarro García, Federico; Arroyo Nombela, Francisco Javier; Román González, Elvira; Martín Brieva, Humberto; Fernández-Acero Bascones, Teresa; Sanz Santamaría, Ana Belén; Díaz Del Toro, Silvia; Calvo De Pablo, Pilar; Patiño Álvarez, Aurora Belén; González Zorn, Bruno; Suárez Rodríguez, Mónica; Goyache Goñi, Joaquín; Escudero García-Calderón, José Antonio; Prieto Prieto, Antonio Daniel; Ugarte Ruiz, María; Gil Serna, JessicaDurante el curso anterior un equipo de la UCM en el marco de un proyecto previo INNOVA-Docencia UCM instauró en España de manera pionera la iniciativa de aprendizaje activo Small World Initiative, de origen norteamericano. Los objetivos de esta iniciativa son (1) Crear cultura científica y acercar la investigación biomédica a niveles educativos en los que los estudiantes tienen aún capacidad de decisión sobre su futura orientación formativa con el fin de fomentar la vocación en I+D; y (2) Promover la concienciación social sobre el uso racional de los antibióticos y la amenaza de la resistencia bacteriana a estos fármacos. En el entorno español se propuso implementar esta estrategia mediante Aprendizaje-Servicio. En esta segunda edición (SWI@UCM 2.0) se ha trabajado en la consolidación, expansión y mejora del proyecto, con énfasis en la integración de los diversos niveles educativos que integran el proyecto (universitario y preuniversitario) y la divulgación científica. Item MicroMundo: Ciencia Ciudadana para el descubrimiento y concienciación sobre el uso de antibióticos mediante Aprendizaje-Servicio(2020) Jiménez Cid, Víctor; Calvo De Pablo, Pilar; Román González, Elvira; Patiño Álvarez, Aurora Belén; González Zorn, Bruno; Gómez-Luz Centellés, María Luisa; Molina Martín, María; Martín Brieva, Humberto; Escudero García-Calderón, José Antonio; Pavón Verges, Mónica; Rodríguez Fernández, Carmina; Díaz Del Toro, Silvia; Rodríguez Escudero, María Isabel; Vázquez Estévez, María Covadonga Inmaculada; Bezos Garrido, Javier; Álvarez Sánchez, Julio; Amaro Torres, Francisco; Suárez Rodríguez, Mónica; Gil Serna, Jessica; Fernández-Acero Bascones, Teresa; Sanz Santamaría, Ana Belén; González Rubio, Gema; Jiménez Gutiérrez, Elena; Coronas Serna, Julia María; Valentí Sanguino, Marta; Del Val Oriza, Elba; Sellers Moya, Ángela; Gibello Prieto, Alicia; Arias López, Patricia; Sastre Vergara, LucíaMicroMundo es un proyecto de Ciencia Ciudadana basado en aprendizaje activo, en el que estudiantes de ESO y Bachillerato, objeto del servicio, coordinados por equipos de estudiantes universitarios, objeto del aprendizaje, desarrollan un proyecto de investigación real. El objetivo de la investigación es el aislamiento, a partir de microorganismos de muestras de suelo, de nuevas actividades antibióticas. Los objetivos de servicio son acercar cultura científica a la sociedad en Salud Global, con foco en el problema de la resistencia a antibióticos, actuando sobre la población más joven,al tiempo que se crean en ellos vocaciones STEM y curiosidad por la I+D en Biomedicina. Durante el curso 2019-20, 150 nuevos alumnos se han incorporado al proyecto interfacultativo en la UCM, trabajando en 26 centros educativos de la Comunidad de Madrid. El confinamiento por la situación pandémica ha reforzado el interés del proyecto, pero ha obligado a pasar el entorno on-line algunas de las actuaciones previstas. No obstante, los objetivos del proyecto se han alcanzado.Item Heterologous mammalian Akt disrupts plasma membrane homeostasis by taking over TORC2 signaling in Saccharomyces cerevisiae.(Scientific Reports, 2018) Rodríguez Escudero, María Isabel; Fernández-Acero Bascones, Teresa; Jiménez Cid, Víctor; Molina, MaríaThe Akt protein kinase is the main transducer of phosphatidylinositol-3,4,5-trisphosphate (PtdIns3,4,5P) signaling in higher eukaryotes, controlling cell growth, motility, proliferation and survival. By co-expression of mammalian class I phosphatidylinositol 3-kinase (PI3K) and Akt in the Saccharomyces cerevisiae heterologous model, we previously described an inhibitory effect on yeast growth that relied on Akt kinase activity. Here we report that PI3K-Akt expression in yeast triggers the formation of large plasma membrane (PM) invaginations that were marked by actin patches, enriched in PtdIns4,5P and associated to abnormal intracellular cell wall deposits. These effects of Akt were mimicked by overproduction of the PtdIns4,5P effector Slm1, an adaptor of the Ypk1 and Ypk2 kinases in the TORC2 pathway. Although Slm1 was phosphorylated in vivo by Akt, TORC2-dependent Ypk1 activation did not occur. However, PI3K-activated Akt suppressed the lethality derived from inactivation of either TORC2 or Ypk protein kinases. Thus, heterologous co-expression of PI3K and Akt in yeast short-circuits PtdIns4,5P- and TORC2-signaling at the level of the Slm-Ypk complex, overriding some of its functions. Our results underscore the importance of phosphoinositide-dependent kinases as key actors in the homeostasis and dynamics of the PM.Item A yeast-based in vivo bioassay to screen for class I phosphatidylinositol 3-kinase specific inhibitors.(Journal of biomolecular screening, 2012) Fernández-Acero Bascones, Teresa; Rodríguez Escudero, María Isabel; Vicente, Francisca; Monteiro, Maria Cândida; Tormo, José R.; Cantizani, Juan; Molina, María; Cid, Víctor J.The phosphatidylinositol 3-kinase (PI3K) pathway couples receptor-mediated signaling to essential cellular functions by generating the lipid second messenger phosphatidylinositol-3,4,5-trisphosphate. This pathway is implicated in multiple aspects of oncogenesis. A low-cost bioassay that readily measures PI3K inhibition in vivo would serve as a valuable tool for research in this field. Using heterologous expression, we have previously reconstituted the PI3K pathway in the model organism Saccharomyces cerevisiae. On the basis of the fact that the overproduction of PI3K is toxic in yeast, we tested the ability of commercial PI3K inhibitors to rescue cell growth. All compounds tested counteracted the PI3K-induced toxicity. Among them, 15e and PI-103 were the most active. Strategies to raise the intracellular drug concentration, specifically the use of 0.003% sodium dodecyl sulfate and the elimination of the Snq2 detoxification pump, optimized the bioassay by enhancing its sensitivity. The humanized yeast-based assay was then tested on a pilot scale for high-throughput screening (HTS) purposes using a collection of natural products of microbial origin. From 9600 extracts tested, 0.6% led to a recovery of yeast growth reproducibly, selectively, and in a dose-dependent manner. Cumulatively, we show that the developed PI3K inhibition bioassay is robust and applicable to large-scale HTS.Item Expression of Human PTEN-L in a Yeast Heterologous Model Unveils Specific N-Terminal Motifs Controlling PTEN-L Subcellular Localization and Function(Cells, 2019) Fernández-Acero Bascones, Teresa; Bertalmio, Eleonora; Luna, Sandra; Mingo, Janire; Bravo Plaza, Ignacio; Rodríguez Escudero, María Isabel; Molina Martín, María; Pulido, Rafael; Jiménez Cid, VíctorThe tumour suppressor PTEN is frequently downregulated, mutated or lost in several types of tumours and congenital disorders including PHTS (PTEN Hamartoma Tumour Syndrome) and ASD (Autism Spectrum Disorder). PTEN is a lipid phosphatase whose activity over the lipid messenger PIP3 counteracts the stimulation of the oncogenic phosphatidylinositol 3-kinase (PI3K) pathway. Recently, several extended versions of PTEN produced in the cell by alternative translation initiation have been described, among which, PTEN-L and PTEN-M represent the longest isoforms. We previously developed a humanized yeast model in which the expression of PI3K in Saccharomyces cerevisiae led to growth inhibition that could be suppressed by co-expression of PTEN. Here, we show that the expression of PTEN-L and PTEN-M in yeast results in robust counteracting of PI3K-dependent growth inhibition. N-terminally tagged GFP-PTEN-L was sharply localized at the yeast plasma membrane. Point mutations of a putative membrane-binding helix located at the PTEN-L extension or its deletion shifted localization to nuclear. Also, a shift from plasma membrane to nucleus was observed in mutants at basic amino acid clusters at the PIP2-binding motif, and at the Cα2 and CBR3 loops at the C2 domain. In contrast, C-terminally tagged PTEN-L-GFP displayed mitochondrial localization in yeast, which was shifted to plasma membrane by removing the first 22 PTEN-L residues. Our results suggest an important role of the N-terminal extension of alternative PTEN isoforms on their spatial and functional regulation.- Project number: 199
Item Propuesta de intervención educativa centrada en el alumno mediante el modelo de clase invertida en la docencia práctica de la asignatura Microbiología Clínica(2021) Román González, Elvira; Pla Alonso, Jesús; Jiménez Cid, Víctor; Fernández-Acero Bascones, Teresa; Prieto Prieto, Antonio Daniel; Rodríguez Escudero, María Isabel; Hidalgo Vico, Susana; Garcia Lavilla, Beatriz; González Rubio, Gema; Pavón Verges, Mónica; Valentí Sanguino, Marta; Román González, Marcos; Sánchez Angulo, ManuelHemos implementado la metodología de clase invertida como modelo de aprendizaje centrado en el alumno en la docencia práctica de la asignatura Microbiología Clínica con el objetivo dinamizar y optimizar el tiempo en el aula. Este tipo de docencia ha favorecido la participación activa del estudiante, su aprendizaje y fomentado un mayor pensamiento crítico que permite la toma de decisiones durante la consecución de las prácticas. Item Yeast-based methods to assess PTEN phosphoinositide phosphatase activity in vivo(Methods, 2015) Rodríguez Escudero, María Isabel; Fernández-Acero Bascones, Teresa; Bravo, Ignacio; Leslie, Nicholas R.; Pulido, Rafael; Molina, María; Jiménez Cid, VíctorThe PTEN phosphoinositide 3-phosphatase is a tumor suppressor commonly targeted by pathologic missense mutations. Subject to multiple complex layers of regulation, its capital role in cancer relies on its counteracting function of class I phosphoinositide 3-kinase (PI3K), a key feature in oncogenic signaling pathways. Precise assessment of the involvement of PTEN mutations described in the clinics in loss of catalytic activity requires either tedious in vitro phosphatase assays or in vivo experiments involving transfection into mammalian cell lines. Taking advantage of the versatility of the model organism Saccharomyces cerevisiae, we have developed different functional assays by reconstitution of the mammalian PI3K-PTEN switch in this lower eukaryote. This methodology is based on the fact that regulated PI3K expression in yeast cells causes conversion of PtdIns(4,5)P2 in PtdIns(3,4,5)P3 and co-expression of PTEN counteracts this effect. This can be traced by monitoring growth, given that PtdIns(4,5)P2 pools are essential for the yeast cell, or by using fluorescent reporters amenable for microscopy or flow cytometry. Here we describe the methodology and review its application to evaluate the functionality of PTEN mutations. We show that the technique is amenable to both directed and systematic structure-function relationship studies, and present an example of its use for the study of the recently discovered PTEN-L variant