Transcriptome-wide RNA processing kinetics revealed using extremely short 4tU labeling.

ABSTRACT:

Background

RNA levels detected at steady state are the consequence of multiple dynamic processes within the cell. In addition to synthesis and decay, transcripts undergo processing. Metabolic tagging with a nucleotide analog is one way of determining the relative contributions of synthesis, decay and conversion processes globally.

Results

By improving 4-thiouracil labeling of RNA in Saccharomyces cerevisiae we were able to isolate RNA produced during as little as 1 minute, allowing the detection of nascent pervasive transcription. Nascent RNA labeled for 1.5, 2.5 or 5 minutes was isolated and analyzed by reverse transcriptase-quantitative polymerase chain reaction and RNA sequencing. High kinetic resolution enabled detection and analysis of short-lived non-coding RNAs as wel

SUBMITTER: Barrass JD 

PROVIDER: S-EPMC4699367 | biostudies-literature | 2015 Dec

REPOSITORIES: biostudies-literature