We are interested in the study of the molecular mechanisms used by plants to modulate their processes of growth and environmental adaptation. We use multidisciplinar approaches such as genetics, biochemistry, physiology, molecular and celular biology with the model plant Arabidopsis thaliana.
Among the plant growth regulators we study the polyamines, molecules that derive from amino acid catabolism and play relevant roles in plant biology. These small aliphatic compounds behave as polications at physiological pH and perform essential functions with unknown molecular mechanisms.
RESEARCH LINES: Our current efforts aim to understand the molecular mechanisms of polyamine action. Several lines of research are carried out in the lab:
- Polyamines and translation. This action aims to the characterization of an exclusive post-translational modification, mediated by the spermidine, of the essential translation factor eIF5A (hypusination). This translation factor, activated by hypusination, is highly conserved among eukaryotes and it has been involved in cell growth, death and autophagy pathways. eIF5A performs diverse actions at the post-transcriptional level (mRNA binding, transport and translation), however its mRNA targets and specific functions remain ill-defined in plants.
Another polyamine that attracts our interest as being involved in translational regulation is the thermospermine, a plant specific polyamine present also in some prokaryotes but absent in animals and fungi. Thermospermine is required for the translation of atypical bHLH factors that present upstream open reading frames (uORFs) in their 5’-leader sequences and are involved in xylem development, although the molecular mechanism is unknown.
- Polyamines and pollen thermotolerance. With this project we want to elucidate the functions of polyamines in pollen germination and pollen tube growth at high temperature. We use the Riboseq technology to identify the translatome features (mRNA sequences translated by ribosomes) of the pollen tube grown in vitro at high temperature conditions. We also study the cross-talk between polyamines and pH on pollen thermotolerance.
- Polyamines and autophagy. This is a recently initiated research line that will be the focus of our future goals. The involvement of spermidine and eIF5A in longevity through the activation of the autophagy process is well documented in animals. However, there is no information with regard to equivalent functions in plants. Our interest is the characterization of the roles of polyamines in the autophagy process in response to nutritional starvation, with the aim to identify new biotechnological targets to improve nitrogen use efficiency and to reduce the use of plant fertilizers.
TECHNOLOGICAL PLATFORMS. To perform the above summarized research lines, we have implemented technologies for protein-protein interactions studies (BiFC) and also for high resolution studies of translation (Ribo-Seq):
- Protein interaction studies by BiFC. We have designed and developed our own binary vectors (http://www.ibmcp.upv.es/FerrandoLabVectors) that facilitate the generation of translational fusions to fluorescent proteins. This set of vectors allow the visualization of protein interactions in vivo in plant cells by means of Bi-molecular-Fluorescence Complementation (BiFC).
- Studies of the translatome by ribosome footprint profiling (Ribo-Seq). We have implemented the advanced technology of Ribo-Seq that allows the global studies of mRNA translation at the sub-codon resolution level, by massive sequencing of ribosome protected mRNA fragments.
Patrick Cottilli; Borja Belda-Palazón; Charith Raj Adkar-Purushothama; Jean-Pierre Perreault; Enrico Schleiff; Ismael Rodrigo; Alejandro Ferrando; Purificación Lisón (2019)Citrus exocortis viroid causes ribosomal stress in tomato plantsNucleic Acids Research. 47 (16): 8649 - 8661
Poidevin L; Unal D; Belda-Palazón B; Ferrando A (2019)Polyamines as Quality Control Metabolites Operating at the Post-Transcriptional Level
Sayas E, Pérez-Benavente B, Manzano C, Farràs R, Alejandro S, Del Pozo JC, Ferrando A, Serrano R. (2018)Polyamines interfere with protein ubiquitylation and cause depletion of intracellular amino acids: a possible mechanism for cell growth inhibition.
FEBS Letters 593(2):209-218
Ferrando A; Castellano MM; Lisón P; Leister D; Stepanova AN; Hanson J (2017)Editorial: Relevance of Translational Regulation on Plant Growth and Environmental ResponsesFrontiers in Plant Science. doi: 10.3389/fpls.20
Belda-Palazon, B., Ferrando, A., and Farràs,R. (2016)Quantitation of protein translation rate in vivo with bioorthogonal Click-chemistry
Methods in Molecular Biology 1449: 369-382
Belda-Palazon B, Almendáriz C, Martí E, Carbonell J, Ferrando A (2016)Relevance of the axis Spermidine/eIF5A for plant growth and developmentFrontiers in Plant Science. 7
Maruri-López I, Hernández-Sánchez IE, Ferrando A, Carbonell J, Jiménez-Bremont JF. (2015)Characterization of maize spermine synthase 1 (ZmSPMS1): evidence for dimerization and intracellular location
Plant Physiology and Biochemistry 97: 264-271
Hernández-Sánchez IE, Maruri-López I, Ferrando A, Carbonell J, Graether SP, Jiménez-Bremont JF (2015)Nuclear localization of the dehydrin OpsDHN1 is determined by histidine-rich domain
Frontiers in Plant Science. Doi: 10.3389/fpls.2015.00702
Belda-Palazón B., Nohales M.A., Rambla J.L., Aceña J.L., Delgado O., Fustero S., Martínez M.C., Granell A., Carbonell J.,Ferrando A. (2014)Biochemical quantiation of the eIF5A hypusination in Arabidopsis thaliana uncovers ABA-dependent regulation.Frontiers in Plant Science 5:202
Belda-Palazón B., Nohales M.A., Rambla J.L., Aceña J.L., Delgado O., Fustero S., Martínez M.C., Granell A., Carbonell J., and Ferrando A. (2014)Biochemical quantiation of the eIF5A hypusination in Arabidopsis thaliana uncovers ABA-dependent regulation.
Frontiers in Plant Science 5:202