High-throughput translational profiling with riboPLATE-seq

Protein synthesis is dysregulated in many diseases, but we lack a systems-level picture of how signaling molecules and RNA binding proteins interact with the translational machinery, largely due to technological limitations. Here we present riboPLATE-seq, a scalable method for generating paired libr...

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Bibliographic Details
Published in:Scientific reports 2022-04, Vol.12 (1), p.5718-5718, Article 5718
Main Authors: Metz, Jordan B., Hornstein, Nicholas J., Sharma, Sohani Das, Worley, Jeremy, Gonzalez, Christian, Sims, Peter A.
Format: Article
Language:English
Subjects:
631/1647/2217/2018
631/337/574
631/45/275
631/61/212/2019
Bar codes
Gene expression
Genomes
High-Throughput Nucleotide Sequencing - methods
Humanities and Social Sciences
Immunoprecipitation
Kinases
mRNA
multidisciplinary
Phosphorylation
Polyribosomes
Polyribosomes - genetics
Polyribosomes - metabolism
Protein Biosynthesis
Protein kinase
Protein synthesis
Proteins
Ribosomes - genetics
Ribosomes - metabolism
RNA-binding protein
rRNA
Science
Science (multidisciplinary)
Sequence Analysis, RNA - methods
Translation
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Summary:Protein synthesis is dysregulated in many diseases, but we lack a systems-level picture of how signaling molecules and RNA binding proteins interact with the translational machinery, largely due to technological limitations. Here we present riboPLATE-seq, a scalable method for generating paired libraries of ribosome-associated and total mRNA. As an extension of the PLATE-seq protocol, riboPLATE-seq utilizes barcoded primers for pooled library preparation, but additionally leverages anti-rRNA ribosome immunoprecipitation on whole polysomes to measure ribosome association (RA). We compare RA to its analogue in ribosome profiling and RNA sequencing, translation efficiency, and demonstrate both the performance of riboPLATE-seq and its utility in detecting translational alterations induced by specific inhibitors of protein kinases.
ISSN:2045-2322
2045-2322
DOI:10.1038/s41598-022-09638-3