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Assessment of Selective mRNA Translation in Mammalian Cells by Polysome Profiling
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Assessment of Selective mRNA Translation in Mammalian Cells by Polysome Profiling

Published on: October 28, 2014

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Mutation at the Site of Hydroxylation in the Ribosomal Protein uL15 (RPL27a) Causes Specific Changes in the

Elizaveta A Zolotenkova1, Alexander V Gopanenko1, Alexey E Tupikin1

  • 1Institute of Chemical Biology and Fundamental Medicine, Siberian Branch of the Russian Academy of Sciences, Novosibirsk 630090, Russia.

International Journal of Molecular Sciences
|April 13, 2023
PubMed

Insights

Ribosomal protein uL15 hydroxylation impacts gene expression. Altering this modification changes which messenger RNAs (mRNAs) are translated, affecting the cell's translatome and transcriptome.

Area of Science:

  • Molecular Biology
  • Cell Biology
  • Genomics

Background:

  • Ribosomal protein uL15 (RPL27a) is hydroxylated at His39, near the tRNA binding site.
  • This hydroxylation modification decreases under hypoxic conditions.
  • Ribosome heterogeneity can influence cellular gene expression.

Purpose of the Study:

  • To investigate the functional impact of RPL27a hydroxylation on mammalian ribosomes.
  • To determine how RPL27a modification affects the translatome and transcriptome.
  • To understand the role of ribosome heterogeneity in mRNA selection.

Main Methods:

  • HEK293T cells were transfected with wild-type or His39Ala mutant RPL27a.
  • RNA-sequencing (RNA-seq) was performed on total cellular and polysome-associated mRNAs.
  • Differentially expressed genes (DEGs) were identified and analyzed for mRNA features.

Main Results:

  • Ribosomes with wild-type and mutant RPL27a were translationally competent.
  • Cells expressing mutant RPL27a showed an enrichment of more abundant and shorter coding sequence (CDS) mRNAs.
  • Cells expressing mutant RPL27a showed a decrease in longer and rarer mRNAs.

Conclusions:

  • Ribosome heterogeneity, influenced by RPL27a modification, alters translatome and transcriptome composition.
  • The hydroxylation status of RPL27a plays a role in selective mRNA translation.
  • This study provides insights into how ribosome properties shape gene expression under varying conditions.
Keywords:
HEK293T cellsRNA-seqRPL27adifferentially expressed genespolysome profilingtranslationuL15 hydroxylation

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