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Characterization of Arabidopsis FPS Isozymes and FPS Gene Expression Analysis Provide Insight into the Biosynthesis of Isoprenoid Precursors in Seeds

Arabidopsis thaliana contains two genes encoding farnesyl diphosphate (FPP) synthase (FPS), the prenyl diphoshate synthase that catalyzes the synthesis of FPP from isopentenyl diphosphate (IPP) and dimethylallyl diphosphate (DMAPP). In this study, we provide evidence that the two Arabidopsis short F...

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Autores principales: Keim, Verónica, Manzano, David, Fernández, Francisco J., Closa, Marta, Andrade, Paola, Caudepón, Daniel, Bortolotti, Cristina, Vega, M. Cristina, Arró, Montserrat, Ferrer, Albert
Formato: Online Artículo Texto
Lenguaje:English
Publicado: Public Library of Science 2012
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC3492304/
https://www.ncbi.nlm.nih.gov/pubmed/23145086
http://dx.doi.org/10.1371/journal.pone.0049109
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author Keim, Verónica
Manzano, David
Fernández, Francisco J.
Closa, Marta
Andrade, Paola
Caudepón, Daniel
Bortolotti, Cristina
Vega, M. Cristina
Arró, Montserrat
Ferrer, Albert
author_facet Keim, Verónica
Manzano, David
Fernández, Francisco J.
Closa, Marta
Andrade, Paola
Caudepón, Daniel
Bortolotti, Cristina
Vega, M. Cristina
Arró, Montserrat
Ferrer, Albert
author_sort Keim, Verónica
collection PubMed
description Arabidopsis thaliana contains two genes encoding farnesyl diphosphate (FPP) synthase (FPS), the prenyl diphoshate synthase that catalyzes the synthesis of FPP from isopentenyl diphosphate (IPP) and dimethylallyl diphosphate (DMAPP). In this study, we provide evidence that the two Arabidopsis short FPS isozymes FPS1S and FPS2 localize to the cytosol. Both enzymes were expressed in E. coli, purified and biochemically characterized. Despite FPS1S and FPS2 share more than 90% amino acid sequence identity, FPS2 was found to be more efficient as a catalyst, more sensitive to the inhibitory effect of NaCl, and more resistant to thermal inactivation than FPS1S. Homology modelling for FPS1S and FPS2 and analysis of the amino acid differences between the two enzymes revealed an increase in surface polarity and a greater capacity to form surface salt bridges of FPS2 compared to FPS1S. These factors most likely account for the enhanced thermostability of FPS2. Expression analysis of FPS::GUS genes in seeds showed that FPS1 and FPS2 display complementary patterns of expression particularly at late stages of seed development, which suggests that Arabidopsis seeds have two spatially segregated sources of FPP. Functional complementation studies of the Arabidopsis fps2 knockout mutant seed phenotypes demonstrated that under normal conditions FPS1S and FPS2 are functionally interchangeable. A putative role for FPS2 in maintaining seed germination capacity under adverse environmental conditions is discussed.
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spelling pubmed-34923042012-11-09 Characterization of Arabidopsis FPS Isozymes and FPS Gene Expression Analysis Provide Insight into the Biosynthesis of Isoprenoid Precursors in Seeds Keim, Verónica Manzano, David Fernández, Francisco J. Closa, Marta Andrade, Paola Caudepón, Daniel Bortolotti, Cristina Vega, M. Cristina Arró, Montserrat Ferrer, Albert PLoS One Research Article Arabidopsis thaliana contains two genes encoding farnesyl diphosphate (FPP) synthase (FPS), the prenyl diphoshate synthase that catalyzes the synthesis of FPP from isopentenyl diphosphate (IPP) and dimethylallyl diphosphate (DMAPP). In this study, we provide evidence that the two Arabidopsis short FPS isozymes FPS1S and FPS2 localize to the cytosol. Both enzymes were expressed in E. coli, purified and biochemically characterized. Despite FPS1S and FPS2 share more than 90% amino acid sequence identity, FPS2 was found to be more efficient as a catalyst, more sensitive to the inhibitory effect of NaCl, and more resistant to thermal inactivation than FPS1S. Homology modelling for FPS1S and FPS2 and analysis of the amino acid differences between the two enzymes revealed an increase in surface polarity and a greater capacity to form surface salt bridges of FPS2 compared to FPS1S. These factors most likely account for the enhanced thermostability of FPS2. Expression analysis of FPS::GUS genes in seeds showed that FPS1 and FPS2 display complementary patterns of expression particularly at late stages of seed development, which suggests that Arabidopsis seeds have two spatially segregated sources of FPP. Functional complementation studies of the Arabidopsis fps2 knockout mutant seed phenotypes demonstrated that under normal conditions FPS1S and FPS2 are functionally interchangeable. A putative role for FPS2 in maintaining seed germination capacity under adverse environmental conditions is discussed. Public Library of Science 2012-11-07 /pmc/articles/PMC3492304/ /pubmed/23145086 http://dx.doi.org/10.1371/journal.pone.0049109 Text en © 2012 Keim et al http://creativecommons.org/licenses/by/4.0/ This is an open-access article distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are properly credited.
spellingShingle Research Article
Keim, Verónica
Manzano, David
Fernández, Francisco J.
Closa, Marta
Andrade, Paola
Caudepón, Daniel
Bortolotti, Cristina
Vega, M. Cristina
Arró, Montserrat
Ferrer, Albert
Characterization of Arabidopsis FPS Isozymes and FPS Gene Expression Analysis Provide Insight into the Biosynthesis of Isoprenoid Precursors in Seeds
title Characterization of Arabidopsis FPS Isozymes and FPS Gene Expression Analysis Provide Insight into the Biosynthesis of Isoprenoid Precursors in Seeds
title_full Characterization of Arabidopsis FPS Isozymes and FPS Gene Expression Analysis Provide Insight into the Biosynthesis of Isoprenoid Precursors in Seeds
title_fullStr Characterization of Arabidopsis FPS Isozymes and FPS Gene Expression Analysis Provide Insight into the Biosynthesis of Isoprenoid Precursors in Seeds
title_full_unstemmed Characterization of Arabidopsis FPS Isozymes and FPS Gene Expression Analysis Provide Insight into the Biosynthesis of Isoprenoid Precursors in Seeds
title_short Characterization of Arabidopsis FPS Isozymes and FPS Gene Expression Analysis Provide Insight into the Biosynthesis of Isoprenoid Precursors in Seeds
title_sort characterization of arabidopsis fps isozymes and fps gene expression analysis provide insight into the biosynthesis of isoprenoid precursors in seeds
topic Research Article
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC3492304/
https://www.ncbi.nlm.nih.gov/pubmed/23145086
http://dx.doi.org/10.1371/journal.pone.0049109
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