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Real-Time Observation of Transcription Initiation and Elongation on an Endogenous Yeast Gene
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Real-Time Observation of Transcription Initiation and Elongation on an Endogenous Yeast Gene
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Journal Article
Real-Time Observation of Transcription Initiation and Elongation on an Endogenous Yeast Gene
2011
Overview
Cellular messenger RNA levels are achieved by the combinatorial complexity of factors controlling transcription, yet the small number of molecules involved in these pathways fluctuates stochastically. It has not yet been experimentally possible to observe the activity of single polymerases on an endogenous gene to elucidate how these events occur in vivo. Here, we describe a method of fluctuation analysis of fluorescently labeled RNA to measure dynamics of nascent RNA—including initiation, elongation, and termination—at an active yeast locus. We find no transcriptional memory between initiation events, and elongation speed can vary by threefold throughout the cell cycle. By measuring the abundance and intranuclear mobility of an upstream transcription factor, we observe that the gene firing rate is directly determined by trans-activating factor search times.
Publisher
American Association for the Advancement of Science,The American Association for the Advancement of Science
Subject
Adenosine Triphosphatases - genetics
/ ATPases Associated with Diverse Cellular Activities
/ Biological and medical sciences
/ Fundamental and applied biological sciences. Psychology
/ Genes
/ Glutamate Synthase - genetics
/ Kinetics
/ loci
/ Molecular and cellular biology
/ RNA
/ RNA Polymerase II - metabolism
/ RNA, Messenger - biosynthesis
/ Saccharomyces cerevisiae - genetics
/ Saccharomyces cerevisiae - metabolism
/ Saccharomyces cerevisiae Proteins - genetics
/ Saccharomyces cerevisiae Proteins - metabolism
/ Transcription Factors - metabolism
/ Transcription. Transcription factor. Splicing. Rna processing
/ Yeast
/ Yeasts
