Large-Scale Measurement of mRNA Degradation in Escherichia coli: To Delay or Not to Delay

Methods Enzymol. 2018:612:47-66. doi: 10.1016/bs.mie.2018.07.003. Epub 2018 Aug 29.

Abstract

In this study, we compared different computational methods used for genome-wide determination of mRNA half-lives in Escherichia coli with a special focus on the impact on considering a delay before the onset of mRNA decay after transcription arrest. A wide variety of datasets were analyzed coming from different technical methods for mRNA quantification (microarrays, RNA-seq, and RT-qPCR) and different bacterial growth conditions. The exponential decay of mRNA levels was fitted using both linear and exponential models and with or without a delay. We showed that for all the models, independently of mRNA quantification methods and growth conditions, ignoring the delay resulted in only a modest overestimation of the half-life. For approximately 80% of the mRNAs, differences in mRNA half-life values were less than 34s. The correlation between half-lives estimated with and without a delay was extremely high. However, the slope of the linear regression between the half-lives with and without a delay tended to decrease with the delay. For the few mRNAs for which taking into account the delay influenced the estimated half-life, the impact was dependent on the model and the growth condition. The smallest impact was obtained for the linear model.

Keywords: Delay in the degradation kinetics; Exponential decay; Genome-wide measurement; RNA decay modeling; RNA half-life.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Escherichia coli / genetics*
  • RNA Stability / genetics
  • RNA Stability / physiology*
  • RNA, Bacterial / metabolism*
  • RNA, Messenger / metabolism*
  • Transcription, Genetic / genetics

Substances

  • RNA, Bacterial
  • RNA, Messenger