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Methods Mol Biol. 2014;1158:157-74. doi: 10.1007/978-1-4939-0700-7_10.

Analysis of mRNA translation states in Arabidopsis over the diurnal cycle by polysome microarray.

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1
Department of Biochemistry, Cellular and Molecular Biology, The University of Tennessee, Knoxville, TN, 37996-0840, USA, amissra@utk.edu.

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.

PMID:
24792050
DOI:
10.1007/978-1-4939-0700-7_10
[Indexed for MEDLINE]
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