Implication of polymerase recycling for nascent transcript quantification by live cell imaging.

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Version: Final published version
License: CC BY 4.0
Serval ID
serval:BIB_4DBF20DED918
Type
Article: article from journal or magazin.
Collection
Publications
Institution
Title
Implication of polymerase recycling for nascent transcript quantification by live cell imaging.
Journal
Yeast
Author(s)
Kindongo O., Lieb G., Skaggs B., Dusserre Y., Vincenzetti V., Pelet S.
ISSN
1097-0061 (Electronic)
ISSN-L
0749-503X
Publication state
Published
Issued date
04/2024
Peer-reviewed
Oui
Volume
41
Number
4
Pages
279-294
Language
english
Notes
Publication types: Journal Article
Publication Status: ppublish
Abstract
Transcription enables the production of RNA from a DNA template. Due to the highly dynamic nature of transcription, live-cell imaging methods play a crucial role in measuring the kinetics of this process. For instance, transcriptional bursts have been visualized using fluorescent phage-coat proteins that associate tightly with messenger RNA (mRNA) stem loops formed on nascent transcripts. To convert the signal emanating from a transcription site into meaningful estimates of transcription dynamics, the influence of various parameters on the measured signal must be evaluated. Here, the effect of gene length on the intensity of the transcription site focus was analyzed. Intuitively, a longer gene can support a larger number of transcribing polymerases, thus leading to an increase in the measured signal. However, measurements of transcription induced by hyper-osmotic stress responsive promoters display independence from gene length. A mathematical model of the stress-induced transcription process suggests that the formation of gene loops that favor the recycling of polymerase from the terminator to the promoter can explain the observed behavior. One experimentally validated prediction from this model is that the amount of mRNA produced from a short gene should be higher than for a long one as the density of active polymerase on the short gene will be increased by polymerase recycling. Our data suggest that this recycling contributes significantly to the expression output from a gene and that polymerase recycling is modulated by the promoter identity and the cellular state.
Keywords
Transcription, Genetic, Promoter Regions, Genetic, RNA, Messenger/genetics, MAPK signaling pathways, gene looping, phage‐coat proteins, stress response, transcription dynamics
Pubmed
Web of science
Open Access
Yes
Create date
25/03/2024 15:29
Last modification date
23/04/2024 7:11
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