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Yeast mRNA decay analysis after addition of 3 Âµ/mL thiolutin

ABSTRACT: Time course after the addition of the transcriptional inhibitor thiolutin at 3Âµg/mL to an exponential growing culture of S. cerevisiae in YPD. Total RNA from BQS252 yeast strain (Mat a, ura 3-52 derived from FY1679) growing in exponential phase in YPD was extracted at different times after thiolutin 3Âµg/mL addition. All the samples were hybridized on the same nylon array. In order to estimate mRNA stability the intensities where normalized using global poly(A) data from a RNA dot-blot and assuming a first order decaying. There is only 1 replicate.

ORGANISM(S): Saccharomyces cerevisiae

SUBMITTER: JosÃ©.E. PÃ©rez-OrtÃn

PROVIDER: E-GEOD-7261 | biostudies-arrayexpress |

REPOSITORIES: biostudies-arrayexpress

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The transcriptional inhibitor thiolutin blocks mRNA degradation in yeast.

Pelechano Vicent V Pérez-Ortín José E JE

Yeast (Chichester, England) 20080201 2

Thiolutin is commonly used as a general inhibitor of transcription in yeast. It has been used to calculate mRNA decay rates by stopping the transcription and then determining the relative abundance of individual mRNAs at different times after inhibition. We report here that thiolutin is also an inhibitor of mRNA degradation, and thus its use can lead to miscalculations of mRNA half-lives. The inhibition of mRNA decay seems to affect the mRNA degradation pathway without impeding poly(A) shortenin ...[more]

PMID: 17914747

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