Cycloheximide can distort measurements of mRNA levels and translation efficiency

Daniel A. Santos, Lei Shi, Benjamin P Tu, Jonathan S. Weissman

Research output: Contribution to journalArticle

3 Citations (Scopus)

Abstract

Regulation of the efficiency with which an mRNA is translated into proteins represents a key mechanism for controlling gene expression. Such regulation impacts the number of actively translating ribosomes per mRNA molecule, referred to as translation efficiency (TE), which can be monitored using ribosome profiling and RNA-seq, or by evaluating the position of an mRNA in a polysome gradient. Here we show that in budding yeast, under nutrient limiting conditions, the commonly used translation inhibitor cycloheximide induces rapid transcriptional upregulation of hundreds of genes involved in ribosome biogenesis. Cycloheximide also prevents translation of these newly transcribed messages, leading to an apparent drop in TE of these genes under conditions that include key transitions during the yeast metabolic cycle, meiosis, and amino acid starvation; however, this effect is abolished when cycloheximide pretreatment is omitted. This response requires TORC1 signaling, and is modulated by the genetic background as well as the vehicle used to deliver the drug. The present work highlights an important caveat to the use of translation inhibitors when measuring TE or mRNA levels, and will hopefully aid in future experimental design as well as interpretation of prior results.

Original languageEnglish (US)
Pages (from-to)4974-4985
Number of pages12
JournalNucleic acids research
Volume47
Issue number10
DOIs
StatePublished - Jun 4 2019

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Protein Biosynthesis
Cycloheximide
Ribosomes
Messenger RNA
Saccharomycetales
Polyribosomes
Meiosis
Starvation
Genes
Research Design
Up-Regulation
Yeasts
RNA
Gene Expression
Amino Acids
Food
Pharmaceutical Preparations
Proteins

ASJC Scopus subject areas

  • Genetics

Cite this

Cycloheximide can distort measurements of mRNA levels and translation efficiency. / Santos, Daniel A.; Shi, Lei; Tu, Benjamin P; Weissman, Jonathan S.

In: Nucleic acids research, Vol. 47, No. 10, 04.06.2019, p. 4974-4985.

Research output: Contribution to journalArticle

Santos, Daniel A. ; Shi, Lei ; Tu, Benjamin P ; Weissman, Jonathan S. / Cycloheximide can distort measurements of mRNA levels and translation efficiency. In: Nucleic acids research. 2019 ; Vol. 47, No. 10. pp. 4974-4985.
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