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

Détails

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Etat: Public
Version: Final published version
Licence: CC BY 4.0
ID Serval
serval:BIB_4DBF20DED918
Type
Article: article d'un périodique ou d'un magazine.
Collection
Publications
Institution
Titre
Implication of polymerase recycling for nascent transcript quantification by live cell imaging.
Périodique
Yeast
Auteur⸱e⸱s
Kindongo O., Lieb G., Skaggs B., Dusserre Y., Vincenzetti V., Pelet S.
ISSN
1097-0061 (Electronic)
ISSN-L
0749-503X
Statut éditorial
Publié
Date de publication
04/2024
Peer-reviewed
Oui
Volume
41
Numéro
4
Pages
279-294
Langue
anglais
Notes
Publication types: Journal Article
Publication Status: ppublish
Résumé
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.
Mots-clé
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
Oui
Création de la notice
25/03/2024 15:29
Dernière modification de la notice
23/04/2024 7:11
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