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2017 ; 45
(3
): e15
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Analysis of translation using polysome profiling
#MMPMID28180329
Chassé H
; Boulben S
; Costache V
; Cormier P
; Morales J
Nucleic Acids Res
2017[Feb]; 45
(3
): e15
PMID28180329
show ga
During the past decade, there has been growing interest in the role of
translational regulation of gene expression in many organisms. Polysome profiling
has been developed to infer the translational status of a specific mRNA species
or to analyze the translatome, i.e. the subset of mRNAs actively translated in a
cell. Polysome profiling is especially suitable for emergent model organisms for
which genomic data are limited. In this paper, we describe an optimized protocol
for the purification of sea urchin polysomes and highlight the critical steps
involved in polysome purification. We applied this protocol to obtain
experimental results on translational regulation of mRNAs following
fertilization. Our protocol should prove useful for integrating the study of the
role of translational regulation in gene regulatory networks in any biological
model. In addition, we demonstrate how to carry out high-throughput processing of
polysome gradient fractions, for the simultaneous screening of multiple
biological conditions and large-scale preparation of samples for next-generation
sequencing.
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