Regulation of Mammalian Messenger RNA Stability Via the Open Reading Frame

Released on by Megan E. Forrest
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  • Regulation of Mammalian Messenger RNA Stability Via the Open Reading Frame Book Detail

  • Author : Megan E. Forrest
  • Release Date : 2020
  • Publisher :
  • Genre : Gene expression
  • Pages : 172
  • ISBN 13 :
  • File Size : 28,28 MB
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Regulation of Mammalian Messenger RNA Stability Via the Open Reading Frame by Megan E. Forrest PDF Summary

Book Description: The role of mRNA decay in dynamic control of gene expression has long been appreciated, where changes in mRNA stability in response to various cellular conditions can have widespread effects on steady-state mRNA levels. While mRNA stability is controlled in part by ribonucleoprotein composition, the primary codon sequence within the open reading frame has increasingly been recognized as an important determinant of stability in eukaryotes, particularly with regard to differential utilization and incorporation of codons by the translation machinery. The data presented in this dissertation expand upon previous studies linking differences in codon composition in the mRNA open reading frame to effects on translation and stability in budding yeast (S. cerevisiae). The principle aim of this research was to characterize the effects of open reading frame sequence on mRNA decay rates in mammalian cells. Using a combination of single-gene mRNA reporters and transcriptome-wide half-life determinations by metabolic labeling in human (HeLa) and Chinese hamster ovary cells, we determined that both synonymous codon content and encoded amino acid sequence are determinants of mRNA stability in mammalian cells, in contrast to previous studies in other model organisms. Further, the relative codon-specific stabilities observed in HeLa were found to be associated with both cognate tRNA availability and intracellular amino acid levels using empirical measurements of tRNA and amino acid levels. Accordingly, codon and amino acid content were found to contribute significantly to the observed variation in half-lives for endogenous mRNAs within functionally related gene groups. Further, both codon and amino acid-based effects on stability were found to vary among different organisms, supporting the idea that open reading frame-based effects on stability differ throughout evolution.The novel finding that amino acid content within a given mRNA transcript contributes to its stability raises the possibility of control of gene expression regulation in response to changes in metabolic status, which can be applied towards a better understanding of various disease mechanisms. Collectively, these results also have implications for improving our understanding of general mRNA decay mechanisms in mammals, highlighting important potential differences between yeast and higher eukaryotes at the level of mRNA sequence decoding that merit further exploration.

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