Allele-Selective Transcriptome Recruitment to Polysomes Primed for Translation: Protein-Coding and Noncoding RNAs, and RNA Isoforms

PloS One
Roshan MascarenhasWolfgang Sadée

Abstract

mRNA translation into proteins is highly regulated, but the role of mRNA isoforms, noncoding RNAs (ncRNAs), and genetic variants remains poorly understood. mRNA levels on polysomes have been shown to correlate well with expressed protein levels, pointing to polysomal loading as a critical factor. To study regulation and genetic factors of protein translation we measured levels and allelic ratios of mRNAs and ncRNAs (including microRNAs) in lymphoblast cell lines (LCL) and in polysomal fractions. We first used targeted assays to measure polysomal loading of mRNA alleles, confirming reported genetic effects on translation of OPRM1 and NAT1, and detecting no effect of rs1045642 (3435C>T) in ABCB1 (MDR1) on polysomal loading while supporting previous results showing increased mRNA turnover of the 3435T allele. Use of high-throughput sequencing of complete transcript profiles (RNA-Seq) in three LCLs revealed significant differences in polysomal loading of individual RNA classes and isoforms. Correlated polysomal distribution between protein-coding and non-coding RNAs suggests interactions between them. Allele-selective polysome recruitment revealed strong genetic influence for multiple RNAs, attributable either to differential expre...Continue Reading

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Apr 14, 2017·Viruses·Magdalena Regina Kubiak, Izabela Makałowska
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Datasets Mentioned

BETA
GM10852

Methods Mentioned

BETA
RNA-Seq
RNA seq
transfection
density gradient fractionation
transfections
PCR
electrophoresis

Software Mentioned

DESeq R
miRanalyzer
GENCODE
bowtie2
NuGen
Cufflinks
cuffdiff
samtools
GraphPad
R

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