Real-Time Messenger RNA Dynamics in Bacillus subtilis

Messenger RNA molecules have been localized to different positions in cells and have been followed by time-lapse microscopy. We have used MS2-mVenus–labeled mRNA and single-particle tracking to obtain information on the dynamics of single-mRNA molecules in real time. Using single-molecule tracking,...

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Autores principales:	Laura Sattler, Peter L. Graumann
Formato:	article
Lenguaje:	EN
Publicado:	Frontiers Media S.A. 2021
Materias:	single-molecule tracking mRNA dynamics Bacillus subtilis translation ribosome dynamics Microbiology QR1-502
Acceso en línea:	https://doaj.org/article/2408fd86135b4c41b0d4bb0bf4ffd01d
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spelling	oai:doaj.org-article:2408fd86135b4c41b0d4bb0bf4ffd01d2021-11-18T08:47:04ZReal-Time Messenger RNA Dynamics in Bacillus subtilis1664-302X10.3389/fmicb.2021.760857https://doaj.org/article/2408fd86135b4c41b0d4bb0bf4ffd01d2021-11-01T00:00:00Zhttps://www.frontiersin.org/articles/10.3389/fmicb.2021.760857/fullhttps://doaj.org/toc/1664-302XMessenger RNA molecules have been localized to different positions in cells and have been followed by time-lapse microscopy. We have used MS2-mVenus–labeled mRNA and single-particle tracking to obtain information on the dynamics of single-mRNA molecules in real time. Using single-molecule tracking, we show that several mRNA molecules visualized via two MS2-binding sites and MS2-mVenus expressed in Bacillus subtilis cells show free diffusion through the entire cell and constrained motion predominantly close to the cell membrane and at the polar regions of the cells. Because constrained motion of mRNAs likely reflects molecules complexed with ribosomes, our data support the idea that translation occurs at sites surrounding the nucleoids. Squared displacement analyses show the existence of at least two distinct populations of molecules with different diffusion constants or possibly of three populations, for example, freely mobile mRNAs, mRNAs in transition complexes, or in complex with polysomes. Diffusion constants between differently sized mRNAs did not differ dramatically and were much lower than that of cytosolic proteins. These data agree with the large size of mRNA molecules and suggest that, within the viscous cytoplasm, size variations do not translate into mobility differences. However, at observed diffusion constants, mRNA molecules would be able to reach all positions within cells in a frame of seconds. We did not observe strong differences in the location of confined motion for mRNAs encoding mostly soluble or membrane proteins, indicating that there is no strong bias for localization of membrane protein-encoding transcripts for the cell membrane.Laura SattlerPeter L. GraumannFrontiers Media S.A.articlesingle-molecule trackingmRNA dynamicsBacillus subtilistranslationribosome dynamicsMicrobiologyQR1-502ENFrontiers in Microbiology, Vol 12 (2021)
institution	DOAJ
collection	DOAJ
language	EN
topic	single-molecule tracking mRNA dynamics Bacillus subtilis translation ribosome dynamics Microbiology QR1-502
spellingShingle	single-molecule tracking mRNA dynamics Bacillus subtilis translation ribosome dynamics Microbiology QR1-502 Laura Sattler Peter L. Graumann Real-Time Messenger RNA Dynamics in Bacillus subtilis
description	Messenger RNA molecules have been localized to different positions in cells and have been followed by time-lapse microscopy. We have used MS2-mVenus–labeled mRNA and single-particle tracking to obtain information on the dynamics of single-mRNA molecules in real time. Using single-molecule tracking, we show that several mRNA molecules visualized via two MS2-binding sites and MS2-mVenus expressed in Bacillus subtilis cells show free diffusion through the entire cell and constrained motion predominantly close to the cell membrane and at the polar regions of the cells. Because constrained motion of mRNAs likely reflects molecules complexed with ribosomes, our data support the idea that translation occurs at sites surrounding the nucleoids. Squared displacement analyses show the existence of at least two distinct populations of molecules with different diffusion constants or possibly of three populations, for example, freely mobile mRNAs, mRNAs in transition complexes, or in complex with polysomes. Diffusion constants between differently sized mRNAs did not differ dramatically and were much lower than that of cytosolic proteins. These data agree with the large size of mRNA molecules and suggest that, within the viscous cytoplasm, size variations do not translate into mobility differences. However, at observed diffusion constants, mRNA molecules would be able to reach all positions within cells in a frame of seconds. We did not observe strong differences in the location of confined motion for mRNAs encoding mostly soluble or membrane proteins, indicating that there is no strong bias for localization of membrane protein-encoding transcripts for the cell membrane.
format	article
author	Laura Sattler Peter L. Graumann
author_facet	Laura Sattler Peter L. Graumann
author_sort	Laura Sattler
title	Real-Time Messenger RNA Dynamics in Bacillus subtilis
title_short	Real-Time Messenger RNA Dynamics in Bacillus subtilis
title_full	Real-Time Messenger RNA Dynamics in Bacillus subtilis
title_fullStr	Real-Time Messenger RNA Dynamics in Bacillus subtilis
title_full_unstemmed	Real-Time Messenger RNA Dynamics in Bacillus subtilis
title_sort	real-time messenger rna dynamics in bacillus subtilis
publisher	Frontiers Media S.A.
publishDate	2021
url	https://doaj.org/article/2408fd86135b4c41b0d4bb0bf4ffd01d
work_keys_str_mv	AT laurasattler realtimemessengerrnadynamicsinbacillussubtilis AT peterlgraumann realtimemessengerrnadynamicsinbacillussubtilis
_version_	1718421310576525312

Real-Time Messenger RNA Dynamics in Bacillus subtilis

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