Departament de

Bioquímica i Biologia Molecular (Farmàcia)

Grup de Recerca

PLANT MOLECULAR BIOLOGY

Director

ALBERT FERRER PRATS

e-mail: albertferrer@ub.edu

Tel.: (34) 93.403.44.97  -  Fax: (34) 93.402.45.20

Investigación

        Los isoprenoides vegetales constituyen un grupo muy amplio y heterogéneo de compuestos que desempeñan funciones celulares básicas y tienen numerosas aplicaciones biotecnológicas. Las plantas sintetizan isoprenoides a través de dos vías metabólicas situadas en compartimentos celulares distintos. La vía del ácido mevalónico, que suministra isopentenildifosfato (IPP) para fabricar isoprenoides en el citosol/retículo endoplasmático y en las mitocondrias, y la vía del metileritritol-4-fosfato que proporciona isopentenildifosfato para sintetizar isoprenoides en los plástidos. Nuestro interés se centra en el estudio de la organización y la regulación de la vía del ácido mevalónico en la planta modelo Arabidopsis thaliana. Para ello, estamos caracterizando las enzimas HMG-CoA reductasa, IPP isomerasa, farnesildifosfato sintasa y escualeno sintasa. Estas enzimas participan en la formación de escualeno, el primer intermediario específico de la vía de síntesis de fitoesteroles, que son los productos finales mayoritarios de la vía.

También estamos investigando las bases bioquímicas y moleculares de los circuitos reguladores implicados en el mantenimiento de la homeostasia intracelular de lípidos en plantas y la función de la proteína Arvp en este proceso. El metabolismo de los lípidos es un complejo entramado de vías metabólicas que deben regularse de forma muy precisa para mantener en todo momento la homeostasis celular de las distintas especies lipídicas. Estudios previos en células animales y en levaduras han demostrado la existencia de una regulación coordinada del metabolismo de los fitoesteroles y los esfingolípidos, y hay evidencias de que en las plantas el metabolismo de estos lípidos también está coordinado. En este contexto, se ha propuesto que Arvp puede ser un regulador común del metabolismo de esteroles y esfingolípidos, aunque se desconoce cual es su función concreta. Con este proyecto pretendemos comenzar a dar respuesta a una de las preguntas básicas relacionadas con el metabolismo lipídico en plantas: ¿Cómo se mantiene la homeostasis intracelular de esteroles y esfingolípidos y cómo interaccionan las respectivas vías de síntesis a fin de coordinar sus actividades?. 

Publicaciones recientes: 

Lluch, M.A., Masferrer, A., Arró, M, Boronat, A., Ferrer, A. (2000) Molecular cloning and expression analysis of the mevalonate kinase gene from Arabidopsis thaliana. Plant Mol. Biol., 42: 365-376.

Cunillera, N., Boronat, A., Ferrer, A. (2000) Spatial and temporal patterns of expression directed by the 5' regions of the Arabidopsis thaliana farnesyl diphosphate synthase genes FPS1 and FPS2. Plant Mol. Biol., 44: 747-758.

Cunillera, N., Arró, M., Forés, O., Manzano. D., Ferrer, A. (2000) Characterization of dehydrodolichyl diphosphate synthase of Arabidopsis thaliana, a key enzyme in dolichol biosynthesis. FEBS Lett. 477: 170-174.

Pujol, G., Baskin, T., Casamayor, A., Cortadellas, N., Ferrer, A., Ariño, J. (2000) The Arabidopsis thaliana PPX/PP4 phosphatases: molecular cloning and structural organization of the genes, and immunolocalization of the proteins to plastids. Plant Mol. Biol. 44, 499-511.

Carretero-Paulet, L., Ahumada, I., Cunillera, N., Rodríguez-Concepción, M., Ferrer, A., Boronat, A., Campos, N. (2002) Expression and molecular analysis of the Arabidopsis DXR gene encoding 1-Deoxy-D-Xylulose 5-phosphate reductoisomerase, the first committed enzyme of the 2-C-methyl-D-erythritol 4-phosphate pathway. Plant Physiol. 129, 1581-1591.

Köhle, A., Sommer, S., Yazaki, K., Ferrer, A., Boronat, A., Li, S-M., Heide, L. (2002) High level expression of chorismate pyruvate-lyase (UbiC) and HMG-CoA reductase in hairy root cultures of Lithospermum erythrorhyzon. Plant Cell Physiol. 43, 894-902.

Masferrer, A., Arró, M., Manzano, D., Schaller, H., Fernández-Busquets, X., Moncaleán, P., Fernández, B., Cunillera, N., Boronat, A., Ferrer. A. (2002) Overexpression of Arabidopsis thaliana farnesyldiphosphate synthase (FPS1S) in transgenic Arabidopsis induces a cell death/senescence-like response and reduced cytokinin levels. Plant J. 30, 123-132.

Rodríguez-Concepción, M., Forés, O., Martínez-García, J.F., González, V., Phillips, M.A., Ferrer, A., Boronat, A. (2004) Distinct light-mediated pathways regulate isoprenoid metabolism during seedling development in Arabidopsis thaliana. Plant Cell 16 : 144-156.

Manzano, D., Fernández-Busquets, X., Schaller, H., González, V., Boronat, A., Arró, M.; Ferrer, A. (2004) The metabolic imbalance underlying lesion formation in A. thaliana plants overexpressing farnesyl diphosphate synthase (isoform 1S) leads to oxidative stress and is triggered by the developmental decline of endogenous HMGR activity. Planta, 219: 982-992

Leivar, P., González, V.M., Castel, S., Trelease, R.N., López-Iglesias, C., Arró, M., Boronat, A., Campos, N., Ferrer, A., Fernández-Busquets, X. (2005) Subcellular localization of Arabidopsis 3-hydroxy-3-methylglutaryl-Coenzyme A Reductase. Plant Physiol. 137: 57-69

Manzano, D., Busquets, A., Closa, M., Hoyerová, K., Schaller, H., Kamínek, M., Arró, A., Ferrer, A. (2006) Overexpression of farnesyl diphosphate synthase in Arabidopsis mitochondria triggers light-dependent lesion formation and alters cytokinin homeostasis. Plant Mol. Biol. 61: 195-213

Forés, O., Arró, M., Pahissa, A., Ferrero, S., Germann, M., Stukey, J., McDonough, V., Nickels, J.T., Campos, N., Ferrer, A. (2006) Arabidopsis thaliana expresses two functional isoforms of Arvp, a protein involved in the regulation of cellular lipid homeostasis. Biochim. Biophys. Acta-Mol. Cell Biol. Lipids. 1761: 725-735

Martin, D., Piulachs, M.D., Cunillera, N., Ferrer, A., Bellés, X. (2007) Mitochondrial targeting of farnesyl diphosphate synthase is a widespread phenomenon in eukaryotes. Biochim. Biophys. Acta-Mol. Cell Res. In press (doi:10.1016/j.bbamcr.2006.11.015)

Research interest

         Isoprenoids constitute the largest family of natural products, and most of them are synthesized by plants, where they play basic roles for growth and survival. Many isoprenoids also possess biological properties that are beneficial to humans. Plants utilise two different pathways to synthesize isopentenyl diphosphate (IPP), the universal precursor of all isoprenoid compounds. The mevalonic acid (MVA) pathway, which produces IPP from acetyl-CoA in the cytosol/endoplasmic reticulum compartment, and the 2-C-methyl-D-erythritol 4-phosphate pathway, which produces IPP and dimethylallyl diphosphate in the plastids from glyceraldehyde 3-phosphate and pyruvate.  Cytosolic and mitochondrial isoprenoids are both formed from IPP produced through the MVA pathway, whereas plastidial isoprenoids are synthesised from IPP provided by the MEP pathway. Our research interest is focusing in the study of the organization and the regulation of the MVA pathway in the model plant Arabidopsis thaliana. We are currently characterizing a series of key enzymes involved in the synthesis of squalene, which is the first committed precursor for the production of sterols. These compounds are the major end-products synthesized through the MVA pathway. Particularly, we are studying the enzymes HMG-CoA reductase, IPP isomerase, farnesyldiphosphate synthase and squalene synthase.

Lipid metabolism represents a very intricate network of pathways, which has to be regulated in a very precise manner in order to maintain lipid homeostasis within a cell. Previous studies in animal and yeast cells support the existence of a coordinated regulation of the metabolism of sterols and sphingolipids. Some recent findings support the view that the metabolism of these two types of lipids is also coordinately regulated in plant cells. The recently isolated yeast Arv protein may function as a common dual regulator of both sterol and sphingolipid metabolism, although its biological function is still unknown. To elucidate how plant cells achieve and maintain sterol and sphingolipid homeostasis we are currently characterizing Arv proteins from Arabidopsis thaliana and investigating the regulatory circuits that coordinate the synthesis of sterols and sphingolipids in response to changes in the flux of each pathway. This project aims to begin to answer one of the basic questions being formulated by the scientific community in regard to the metabolism of lipids in plants: How is the cellular homeostasis of sterols and sphingolipids maintained and how do the respective synthesis pathways interact to coordinate their activity?. 

Recent publications: 

Lluch, M.A., Masferrer, A., Arró, M, Boronat, A., Ferrer, A. (2000) Molecular cloning and expression analysis of the mevalonate kinase gene from Arabidopsis thaliana. Plant Mol. Biol., 42: 365-376.

Cunillera, N., Boronat, A. and Ferrer, A. (2000) Spatial and temporal patterns of expression directed by the 5' regions of the Arabidopsis thaliana farnesyl diphosphate synthase genes FPS1 and FPS2. Plant Mol. Biol., 44: 747-758.

Cunillera, N., Arró, M., Forés, O., Manzano. D., and Ferrer, A. (2000) Characterization of dehydrodolichyl diphosphate synthase of Arabidopsis thaliana, a key enzyme in dolichol biosynthesis. FEBS Lett. 477: 170-174.

Pujol, G., Baskin, T., Casamayor, A., Cortadellas, N., Ferrer, A., Ariño, J. (2000) The Arabidopsis thaliana PPX/PP4 phosphatases: molecular cloning and structural organization of the genes, and immunolocalization of the proteins to plastids. Plant Mol. Biol. 44, 499-511.

Carretero-Paulet, L., Ahumada, I., Cunillera, N., Rodríguez-Concepción, M., Ferrer, A., Boronat, A., Campos, N. (2002) Expression and molecular analysis of the Arabidopsis DXR gene encoding 1-Deoxy-D-Xylulose 5-phosphate reductoisomerase, the first committed enzyme of the 2-C-methyl-D-erythritol 4-phosphate pathway. Plant Physiol. 129, 1581-1591.

Köhle, A., Sommer, S., Yazaki, K., Ferrer, A., Boronat, A., Li, S-M., Heide, L. (2002) High level expression of chorismate pyruvate-lyase (UbiC) and HMG-CoA reductase in hairy root cultures of Lithospermum erythrorhyzon. Plant Cell Physiol. 43, 894-902.

Masferrer, A., Arró, M., Manzano, D., Schaller, H., Fernández-Busquets, X., Moncaleán, P., Fernández, B., Cunillera, N., Boronat, A., Ferrer. A. (2002) Overexpression of Arabidopsis thaliana farnesyldiphosphate synthase (FPS1S) in transgenic Arabidopsis induces a cell death/senescence-like response and reduced cytokinin levels. Plant J. 30, 123-132.

Rodríguez-Concepción, M., Forés, O., Martínez-García, J.F, González, V., Phillips, M.A., Ferrer, A.; Boronat, A. (2004) Distinct light-mediated pathways regulate isoprenoid metabolism during seedling development in Arabidopsis thaliana. Plant Cell 16 : 144-156.

Manzano, D., Fernández-Busquets, X., Schaller, H., González, V., Boronat, A., Arró, M.; Ferrer, A. (2004) The metabolic imbalance underlying lesion formation in A. thaliana plants overexpressing farnesyl diphosphate synthase (isoform 1S) leads to oxidative stress and is triggered by the developmental decline of endogenous HMGR activity. Planta, 219: 982-992

Leivar, P., González, V.M., Castel, S., Trelease, R.N., López-Iglesias, C., Arró, M., Boronat, A., Campos, N., Ferrer, A., Fernàndez-Busquets, X. (2005) Subcellular localization of Arabidopsis 3-hydroxy-3-methylglutaryl-Coenzyme A Reductase. Plant Physiol. 137 : 57-69

Manzano, D., Busquets, A., Closa, M., Hoyerová, K., Schaller, H., Kamínek, M., Arró, A., Ferrer, A. (2006) Overexpression of farnesyl diphosphate synthase in Arabidopsis mitochondria triggers light-dependent lesion formation and alters cytokinin homeostasis. Plant Mol. Biol. 61: 195-213

Forés, O., Arró, M., Pahissa, A., Ferrero, S., Germann, M., Stukey, J., McDonough, V., Nickels, J.T., Campos, N., Ferrer, A. (2006) Arabidopsis thaliana expresses two functional isoforms of Arvp, a protein involved in the regulation of cellular lipid homeostasis. Biochim. Biophys. Acta-Mol. Cell Biol. Lipids. 1761: 725-735

Martin, D., Piulachs, M.D., Cunillera, N., Ferrer, A., Bellés, X. (2007) Mitochondrial targeting of farnesyl diphosphate synthase is a widespread phenomenon in eukaryotes. Biochim. Biophys. Acta-Mol. Cell Res. In press (doi:10.1016/j.bbamcr.2006.11.015)

Recerca
 

           Els isoprenoides vegetals són un grup molt ampli i heterogeni de compostos, que duen a terme funcions cel·lulars bàsiques i tenen nombroses aplicacions biotecnològiques. Les plantes sintetitzen isoprenoides mitjançant dues vies metabòliques situades en compartiments cel·lulars diferents. La via de l’àcid mevalònic, que subministra isopentenildifosfat (IPP) per fabricar isoprenoides en el citosol/reticle endoplasmàtic i les mitocòndries, i la via del metileritritol-4-fosfat que proporciona IPP per la síntesi d’isoprenoides en els plastidis. El nostre interès està centrat en l’estudi de l’organització i la regulació de la via de l’àcid mevalònic en la planta model Arabidopsis thaliana. Estem treballant en la caracterització d’una sèrie d’enzims que participen en la síntesi de l’esqualé, el primer intermediari específic de la síntesis de fitoesterols, que són els productes finals majoritaris de la via del mevalonat. Concretament, estem treballant en la caracterització bioquímica i molecular dels enzims HMG-CoA reductasa, IPP isomerasa, farnesildifosfat sintasa i esqualé sintasa.

            També estem investigant les bases bioquímiques i moleculars dels circuits reguladors implicats en el manteniment de la homeostasi lipídica intracel·lular en les plantes i el paper que juga en aquest procés la proteïna Arvp. El metabolisme lipídic és un entramat molt complex de vies metabòliques que han de regular-se de forma molt precisa per tal de mantenir en tot moment la homeostasi de les diferents espècies lipídiques. Estudis previs en cèl·lules animals i en llevats han demostrat l’existència d’una regulació coordinada del metabolisme dels fitoesterols i els esfingolípids. Hi ha evidències de que en les plantes el metabolisme d’aquests lípids també està coordinat. En aquest context, es proposa que la proteïna Arv pot ser un regulador comú del metabolisme dels esterols i els esfingolípids, tot i que es desconeix quina és la seva funció concreta. Aquest projecte pretén començar a donar resposta a una de les preguntes bàsiques relacionades amb el metabolisme lipídic de les plantas: Com es manté la homeostasi intracel·lular dels esterols i els esfingolípids i com interaccionen les respectives vies de síntesi per regular coordinadament les seves activitats?.

 Publicacions recents

 Lluch, M.A., Masferrer, A., Arró, M, Boronat, A., Ferrer, A. (2000) Molecular cloning and expression analysis of the  mevalonate kinase gene from Arabidopsis thaliana. Plant Mol. Biol., 42: 365-376.

Cunillera, N., Boronat, A., Ferrer, A. (2000) Spatial and temporal patterns of expression directed by the 5' regions of the Arabidopsis thaliana farnesyl diphosphate synthase genes FPS1 and FPS2. Plant Mol. Biol., 44: 747-758.

Cunillera, N., Arró, M., Forés, O., Manzano. D., Ferrer, A. (2000) Characterization of dehydrodolichyl diphosphate synthase of Arabidopsis thaliana, a key enzyme in dolichol biosynthesis. FEBS Lett. 477: 170-174.

Pujol, G., Baskin, T., Casamayor, A., Cortadellas, N., Ferrer, A., Ariño, J. (2000) The Arabidopsis thaliana PPX/PP4 phosphatases: molecular cloning and structural organization of the genes, and immunolocalization of the proteins to plastids. Plant Mol. Biol. 44, 499-511.

Carretero-Paulet, L., Ahumada, I., Cunillera, N., Rodríguez-Concepción, M., Ferrer, A., Boronat, A., Campos, N. (2002) Expression and molecular analysis of the Arabidopsis DXR gene encoding 1-Deoxy-D-Xylulose 5-phosphate reductoisomerase, the first committed enzyme of the 2-C-methyl-D-erythritol 4-phosphate pathway. Plant Physiol. 129, 1581-1591.

Köhle, A., Sommer, S., Yazaki, K., Ferrer, A., Boronat, A., Li, S-M., Heide, L. (2002) High level expression of chorismate pyruvate-lyase (UbiC) and HMG-CoA reductase in hairy root cultures of Lithospermum erythrorhyzon. Plant Cell Physiol. 43, 894-902.

Masferrer, A., Arró, M., Manzano, D., Schaller, H., Fernández-Busquets, X., Moncaleán, P., Fernández, B., Cunillera, N., Boronat, A., Ferrer. A. (2002) Overexpression of Arabidopsis thaliana farnesyldiphosphate synthase (FPS1S) in transgenic Arabidopsis induces a cell death/senescence-like response and reduced cytokinin levels. Plant J. 30, 123-132.

Rodríguez-Concepción, M., Forés, O., Martínez-García, J.F, González, V., Phillips, M.A., Ferrer, A.; Boronat, A. (2004) Distinct light-mediated pathways regulate isoprenoid metabolism during seedling development in Arabidopsis thaliana. Plant Cell 16 : 144-156.

Manzano, D., Fernández-Busquets, X., Schaller, H., González, V., Boronat, A., Arró, M.; Ferrer, A. (2004) The metabolic imbalance underlying lesion formation in A. thaliana plants overexpressing farnesyl diphosphate synthase (isoform 1S) leads to oxidative stress and is triggered by the developmental decline of endogenous HMGR activity. Planta, 219: 982-992

Leivar, P., González, V.M., Castel, S., Trelease, R.N., López-Iglesias, C., Arró, M., Boronat, A., Campos, N., Ferrer, A., Fernàndez-Busquets, X. (2005) Subcellular localization of Arabidopsis 3-hydroxy-3-methylglutaryl-Coenzyme A Reductase. Plant Physiol. 137 : 57-69

Manzano, D., Busquets, A., Closa, M., Hoyerová, K., Schaller, H., Kamínek, M., Arró, A., Ferrer, A. (2006) Overexpression of farnesyl diphosphate synthase in Arabidopsis mitochondria triggers light-dependent lesion formation and alters cytokinin homeostasis. Plant Mol. Biol. 61: 195-213

Forés, O., Arró, M., Pahissa, A., Ferrero, S., Germann, M., Stukey, J., McDonough, V., Nickels, J.T., Campos, N., Ferrer, A. (2006) Arabidopsis thaliana expresses two functional isoforms of Arvp, a protein involved in the regulation of cellular lipid homeostasis. Biochim. Biophys. Acta-Mol. Cell Biol. Lipids. 1761: 725-735

Martin, D., Piulachs, M.D., Cunillera, N., Ferrer, A., Bellés, X. (2007) Mitochondrial targeting of farnesyl diphosphate synthase is a widespread phenomenon in eukaryotes. Biochim. Biophys. Acta-Mol. Cell Res. In press (doi:10.1016/j.bbamcr.2006.11.015)