Paper de l’agmatina i les poliamines en el fracàs renal d’animals diabètics

Projecte de recerca elaborat a partir d’una estada a la University of California, Estats Units entre febrer i maig de 2007. L’arginina és un component de les proteïnes i juga un paper important en respostes inflamatòries. S’ha demostrat que l’agmatina pot inhibir el creixement mitjançant la supressió de la biosíntesi i el transport de poliamines. Actualment s’està avaluant el mecanisme de l’aturada d’aquest creixement. S’està estudiant també l’impacte en l’apoptosi i la senescència, ja que els models apoptòtics impliquen un descens de poliamines com a factor comú i causal d’apoptosi. En la diabetis el ronyó creix i l’índex de filtració glomerular esdevé supranormal. Aquest creixement renal precoç és considerat una resposta compensatòria a l’increment de la càrrega hiperfiltrada. Nosaltres considerem que aquesta hiperfiltració glomerular diabètica es deguda a esdeveniments que ocorren en el creixement del túbul proximal, i en aquest creixement hi poden estar involucrades les poliamines.

Poliamines en estrès abiòtic i mecanismes moleculars

Les poliamines (PAs) putrescina (Put), espermidina (Spd) i espermina (Spm) són mol•lècules policatiòniques de baix pes molecular, presents en els microorganismes, animals i plantes. Les PAs han estat implicades en diversos processos cel•lulars importants, incloent la resposta de les plantes a l'estrès. No obstant això, el seu mode d'acció està es desconeix. En les plantes, es van acumulant evidències de que les PAs interactuen amb macromolècules i estructures cel•lulars, com ara proteïnes de membrana, i la seva possible participació en transducció de senyals s'ha convertit en una creixent i interessant àrea d'estudi. En aquesta tesi, la possible interacció entre les poliamines i les vies de senyalització de fosfolípids és investigada. Resultats previs, han posat de manifest que alteracions en els nivells endògens de poliamines (PAs), per sobreexpressió gènica o pèrdua de funció de gens de biosíntesi, redueixen o milloren, respectivament, la capacitat de les plantes d'Arabidopsis per tolerar agressions per estrès abiòtic, produint en alguns casos notables alteracions en el desenvolupament. En aquestes plantes amb nivells alterats de PAs s'han detectat canvis importants en l'expressió gènica i s'ha trobat una connexió entre el contingut de PAs ii la biosíntesi / senyalització d'àcid abscísic (ABA). La hipòtesi actual de treball és que aquestes alteracions en l'expressió gènica poden estar mediades, si més no en part, pel catabolisme de PAs, i l’acció directa o indirecta de les espècies reactives d'oxigen (ROS) que se’n deriven.

Modulation of plant HMG-CoA reductase by protein phosphatase 2A: Positive and negative control at a key node of metabolism

The enzyme HMG-CoA reductase (HMGR) has a key regulatory role in the mevalonate pathway for isoprenoid biosynthesis, critical not only for normal plant development, but also for the adaptation to demanding environmental conditions. Consistent with this notion, plant HMGR is modulated by many diverse endogenous signals and external stimuli. Protein phosphatase 2A (PP2A) is involved in auxin, abscisic acid, ethylene and brassinosteroid signaling and now emerges as a positive and negative multilevel regulator of plant HMGR, both during normal growth and in response to a variety of stress conditions. The interaction with HMGR is mediated by B" regulatory subunits of PP2A, which are also calcium binding proteins. The new discoveries uncover the potential of PP2A to integrate developmental and calcium-mediated environmental signals in the control of plant HMGR.

Biosynthesis of panaxynol and panaxydol in Panax ginseng

The natural formation of the bioactive C17-polyacetylenes (−)-(R)-panaxynol and panaxydol was analyzed by 13C-labeling experiments. For this purpose, plants of Panax ginseng were supplied with 13CO2 under field conditions or, alternatively, sterile root cultures of P. ginseng were supplemented with [U-13C6]glucose. The polyynes were isolated from the labeled roots or hairy root cultures, respectively, and analyzed by quantitative NMR spectroscopy. The same mixtures of eight doubly 13C-labeled isotopologues and one single labeled isotopologue were observed in the C17-polyacetylenes obtained from the two experiments. The polyketide-type labeling pattern is in line with the biosynthetic origin of the compounds via decarboxylation of fatty acids, probably of crepenynic acid. The 13C-study now provides experimental evidence for the biosynthesis of panaxynol and related polyacetylenes in P. ginseng under in planta conditions as well as in root cultures. The data also show that 13CO2 experiments under field conditions are useful to elucidate the biosynthetic pathways of metabolites, including those from roots.

Biosynthesis of panaxynol and panaxydol in Panax ginseng

The natural formation of the bioactive C17-polyacetylenes (−)-(R)-panaxynol and panaxydol was analyzed by 13C-labeling experiments. For this purpose, plants of Panax ginseng were supplied with 13CO2 under field conditions or, alternatively, sterile root cultures of P. ginseng were supplemented with [U-13C6]glucose. The polyynes were isolated from the labeled roots or hairy root cultures, respectively, and analyzed by quantitative NMR spectroscopy. The same mixtures of eight doubly 13C-labeled isotopologues and one single labeled isotopologue were observed in the C17-polyacetylenes obtained from the two experiments. The polyketide-type labeling pattern is in line with the biosynthetic origin of the compounds via decarboxylation of fatty acids, probably of crepenynic acid. The 13C-study now provides experimental evidence for the biosynthesis of panaxynol and related polyacetylenes in P. ginseng under in planta conditions as well as in root cultures. The data also show that 13CO2 experiments under field conditions are useful to elucidate the biosynthetic pathways of metabolites, including those from roots.

Multilevel control of arabidopsis 3-hydroxy-3-methylglutaryl coenzyme A reductase by protein phosphatase 2A

Plants synthesize a myriad of isoprenoid products that are required both for essential constitutive processes and for adaptive responses to the environment. The enzyme 3-hydroxy-3-methylglutaryl-CoA reductase (HMGR) catalyzes a key regulatory step of the mevalonate pathway for isoprenoid biosynthesis and is modulated by many endogenous and external stimuli. In spite of that, no protein factor interacting with and regulating plant HMGR in vivo has been described so far. Here, we report the identification of two B99 regulatory subunits of protein phosphatase 2A (PP2A), designated B99a and B99b, that interact with HMGR1S and HMGR1L, the major isoforms of Arabidopsis thaliana HMGR. B99a and B99b are Ca2+ binding proteins of the EF-hand type. We show that HMGR transcript, protein, and activity levels are modulated by PP2A in Arabidopsis. When seedlings are transferred to salt-containing medium, B99a and PP2A mediate the decrease and subsequent increase of HMGR activity, which results from a steady rise of HMGR1-encoding transcript levels and an initial sharper reduction of HMGR protein level. In unchallenged plants, PP2A is a posttranslational negative regulator of HMGR activity with the participation of B99b. Our data indicate that PP2A exerts multilevel control on HMGR through the fivemember B99 protein family during normal development and in response to a variety of stress conditions.