Cycloheximide can distort measurements of mRNA levels and translation efficiency.
Cycloheximide
/ pharmacology
Gene Expression Regulation, Fungal
/ drug effects
Meiosis
/ drug effects
Models, Genetic
Polyribosomes
/ metabolism
Protein Biosynthesis
/ drug effects
Protein Synthesis Inhibitors
/ pharmacology
RNA, Messenger
/ genetics
Ribosomes
/ metabolism
Saccharomyces cerevisiae
/ drug effects
Saccharomyces cerevisiae Proteins
/ genetics
Saccharomycetales
/ drug effects
Transcription Factors
/ genetics
Journal
Nucleic acids research
ISSN: 1362-4962
Titre abrégé: Nucleic Acids Res
Pays: England
ID NLM: 0411011
Informations de publication
Date de publication:
04 06 2019
04 06 2019
Historique:
accepted:
16
03
2019
revised:
01
03
2019
received:
16
01
2019
pubmed:
28
3
2019
medline:
26
11
2019
entrez:
28
3
2019
Statut:
ppublish
Résumé
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.
Identifiants
pubmed: 30916348
pii: 5420543
doi: 10.1093/nar/gkz205
pmc: PMC6547433
doi:
Substances chimiques
Protein Synthesis Inhibitors
0
RNA, Messenger
0
Saccharomyces cerevisiae Proteins
0
TORC1 protein complex, S cerevisiae
0
Transcription Factors
0
Cycloheximide
98600C0908
Types de publication
Journal Article
Research Support, N.I.H., Extramural
Research Support, Non-U.S. Gov't
Research Support, U.S. Gov't, Non-P.H.S.
Langues
eng
Sous-ensembles de citation
IM
Pagination
4974-4985Subventions
Organisme : NIGMS NIH HHS
ID : R01 GM094314
Pays : United States
Informations de copyright
© The Author(s) 2019. Published by Oxford University Press on behalf of Nucleic Acids Research.
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