Abstract
Gene regulation at the level of translation occurs in response to environmental perturbation and is increasingly recognized as a factor affecting plant development. Despite extensive knowledge of transcriptional control, very little is known about translational regulation of genes in response to the daily light/dark cycles. Here we describe the experimental layout designed to address how the translation states of genes change at various times during a diurnal cycle in Arabidopsis thaliana seedlings. We have adopted a strategy combining sucrose-gradient profiling of ribosomes and high-throughput microarray analysis of the ribosome-associated mRNA to investigate the translational landscape of the Arabidopsis genome. This is a powerful technique that can be easily extended to study translation regulation in different genetic backgrounds and under various environmental conditions.
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References
Crosthwaite SK, Loros JJ, Dunlap JC (1995) Light-induced resetting of a circadian clock is mediated by a rapid increase in frequency transcript. Cell 81:1003–1012
Dunlap JC (1999) Molecular bases for circadian clocks. Cell 96:271–290
Harmer SL et al (2000) Orchestrated transcription of key pathways in Arabidopsis by the circadian clock. Science 290:2110–2113
Schaffer R et al (2001) Microarray analysis of diurnal and circadian-regulated genes in Arabidopsis. Plant Cell 13:113–123
Edwards KD et al (2006) FLOWERING LOCUS C mediates natural variation in the high-temperature response of the Arabidopsis circadian clock. Plant Cell 18:639–650
Covington MF, Harmer SL (2007) The circadian clock regulates auxin signaling and responses in Arabidopsis. PLoS Biol 5:e222
Michael TP et al (2008) Network discovery pipeline elucidates conserved time-of-day-specific cis-regulatory modules. PLoS Genet 4:e14
Beyer A, Hollunder J, Nasheuer HP, Wilhelm T (2004) Post-transcriptional expression regulation in the yeast Saccharomyces cerevisiae on a genomic scale. Mol Cell Proteomics 3:1083–1092
Tian Q et al (2004) Integrated genomic and proteomic analyses of gene expression in mammalian cells. Mol Cell Proteomics 3:960–969
Piques M et al (2009) Ribosome and transcript copy numbers, polysome occupancy and enzyme dynamics in Arabidopsis. Mol Syst Biol 5:314
Baerenfaller K et al (2012) Systems-based analysis of Arabidopsis leaf growth reveals adaptation to water deficit. Mol Syst Biol 8:606
Kim BH, Cai X, Vaughn JN, von Arnim AG (2007) On the functions of the h subunit of eukaryotic initiation factor 3 in late stages of translation initiation. Genome Biol 8:R60
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This work was supported by NSF grant DBI-0820047.
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Missra, A., von Arnim, A.G. (2014). Analysis of mRNA Translation States in Arabidopsis Over the Diurnal Cycle by Polysome Microarray. In: Staiger, D. (eds) Plant Circadian Networks. Methods in Molecular Biology, vol 1158. Humana Press, New York, NY. https://doi.org/10.1007/978-1-4939-0700-7_10
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DOI: https://doi.org/10.1007/978-1-4939-0700-7_10
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Publisher Name: Humana Press, New York, NY
Print ISBN: 978-1-4939-0699-4
Online ISBN: 978-1-4939-0700-7
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