Ribosomes are a crucial component of the physiological state of a cell.
Therefore, we aimed to monitor ribosome dynamics using a fast and easy
fluorescence readout. Using fluorescent-labeled ribosomal proteins, the
dynamics of ribosomes during batch cultivation and during nutritional
shift conditions was investigated. The fluorescence readout was compared
to the cellular rRNA content determined by capillary gel electrophoresis
with laser-induced fluorescence detection during exponentially
accelerating and decelerating growth. We found a linear correlation
between the observed fluorescence and the extracted rRNA content
throughout cultivation, demonstrating the applicability of this method.
Moreover, the results show that ribosome dynamics, as a result of
slowing growth, are accompanied by the passive effect of dilution of
preexisting ribosomes, de novo ribosome synthesis and ribosome
degradation. In light of the challenging task of deciphering ribosome
regulatory mechanisms, our approach of using fluorescence to follow
ribosome dynamics will allow more comprehensive studies of biological
systems.
%0 Journal Article
%1 ISI:000401757600008
%A Failmezger, Jurek
%A Ludwig, Julian
%A Niess, Alexander
%A Siemann-Herzberg, Martin
%C GREAT CLARENDON ST, OXFORD OX2 6DP, ENGLAND
%D 2017
%I OXFORD UNIV PRESS
%J FEMS MICROBIOLOGY LETTERS
%K myown proteinsynthesis
%N 6
%R 10.1093/femsle/fnx055
%T Quantifying ribosome dynamics in Escherichia coli using fluorescence
%U https://doi.org/10.1093/femsle/fnx055
%V 364
%X Ribosomes are a crucial component of the physiological state of a cell.
Therefore, we aimed to monitor ribosome dynamics using a fast and easy
fluorescence readout. Using fluorescent-labeled ribosomal proteins, the
dynamics of ribosomes during batch cultivation and during nutritional
shift conditions was investigated. The fluorescence readout was compared
to the cellular rRNA content determined by capillary gel electrophoresis
with laser-induced fluorescence detection during exponentially
accelerating and decelerating growth. We found a linear correlation
between the observed fluorescence and the extracted rRNA content
throughout cultivation, demonstrating the applicability of this method.
Moreover, the results show that ribosome dynamics, as a result of
slowing growth, are accompanied by the passive effect of dilution of
preexisting ribosomes, de novo ribosome synthesis and ribosome
degradation. In light of the challenging task of deciphering ribosome
regulatory mechanisms, our approach of using fluorescence to follow
ribosome dynamics will allow more comprehensive studies of biological
systems.
@article{ISI:000401757600008,
abstract = {{Ribosomes are a crucial component of the physiological state of a cell.
Therefore, we aimed to monitor ribosome dynamics using a fast and easy
fluorescence readout. Using fluorescent-labeled ribosomal proteins, the
dynamics of ribosomes during batch cultivation and during nutritional
shift conditions was investigated. The fluorescence readout was compared
to the cellular rRNA content determined by capillary gel electrophoresis
with laser-induced fluorescence detection during exponentially
accelerating and decelerating growth. We found a linear correlation
between the observed fluorescence and the extracted rRNA content
throughout cultivation, demonstrating the applicability of this method.
Moreover, the results show that ribosome dynamics, as a result of
slowing growth, are accompanied by the passive effect of dilution of
preexisting ribosomes, de novo ribosome synthesis and ribosome
degradation. In light of the challenging task of deciphering ribosome
regulatory mechanisms, our approach of using fluorescence to follow
ribosome dynamics will allow more comprehensive studies of biological
systems.}},
added-at = {2018-01-25T13:38:08.000+0100},
address = {{GREAT CLARENDON ST, OXFORD OX2 6DP, ENGLAND}},
affiliation = {{Siemann-Herzberg, M (Reprint Author), Inst Biochem Engn, Allmandring 31, D-70569 Stuttgart, Germany.
Failmezger, Jurek; Ludwig, Julian; Niess, Alexander; Siemann-Herzberg, Martin, Univ Stuttgart, Inst Biochem Engn, D-70569 Stuttgart, Germany.}},
article-number = {{fnx055}},
author = {Failmezger, Jurek and Ludwig, Julian and Niess, Alexander and Siemann-Herzberg, Martin},
author-email = {{siemann@ibvt.uni-stuttgart.de}},
biburl = {https://puma.ub.uni-stuttgart.de/bibtex/238b769217bf2e5f2378519f92013c078/siemannherzberg},
da = {{2018-01-25}},
doc-delivery-number = {{EV4UF}},
doi = {{10.1093/femsle/fnx055}},
eissn = {{1574-6968}},
funding-acknowledgement = {{Bundesministerium fur Bildung und Forschung, Berlin, Germany {[}BMBF]
{[}031A157D]}},
funding-text = {{This work was supported by the Bundesministerium fur Bildung und
Forschung, Berlin, Germany {[}BMBF grant number: 031A157D].}},
interhash = {3b787ec8fa4fd9ba27d4f810a6ddc7e2},
intrahash = {38b769217bf2e5f2378519f92013c078},
issn = {{0378-1097}},
journal = {{FEMS MICROBIOLOGY LETTERS}},
journal-iso = {{FEMS Microbiol. Lett.}},
keywords = {myown proteinsynthesis},
keywords-plus = {{GEL-ELECTROPHORESIS; DEGRADATION; RNA; BACTERIA; GROWTH}},
language = {{English}},
month = {{MAR}},
number = {{6}},
number-of-cited-references = {{20}},
publisher = {{OXFORD UNIV PRESS}},
research-areas = {{Microbiology}},
times-cited = {{3}},
timestamp = {2018-06-14T11:13:43.000+0200},
title = {{Quantifying ribosome dynamics in Escherichia coli using fluorescence}},
type = {{Article}},
unique-id = {{ISI:000401757600008}},
url = {https://doi.org/10.1093/femsle/fnx055},
usage-count-last-180-days = {{4}},
usage-count-since-2013 = {{5}},
volume = {{364}},
web-of-science-categories = {{Microbiology}},
year = {{2017}}
}
