Single-molecule investigations of the stringent response machinery in living bacterial cells
English, Brian P. ; Hauryliuk, Vasili LU
; Sanamrad, Arash
; Tankov, Stoyan
; Dekker, Nynke H.
and Elf, Johan
(2011)
In Proceedings of the National Academy of Sciences of the United States of America
108(31).
p.365-373
- Abstract
The RelA-mediated stringent response is at the heart of bacterial adaptation to starvation and stress, playing a major role in the bacterial cell cycle and virulence. RelA integrates several environmental cues and synthesizes the alarmone ppGpp, which globally reprograms transcription, translation, and replication. We have developed and implemented novel single-molecule tracking methodology to characterize the intracellular catalytic cycle of RelA. Our single-molecule experiments show that RelA is on the ribosome under nonstarved conditions and that the individual enzyme molecule stays off the ribosome for an extended period of time after activation. This suggests that the catalytically active part of the RelA cycle is performed off,... (More)
The RelA-mediated stringent response is at the heart of bacterial adaptation to starvation and stress, playing a major role in the bacterial cell cycle and virulence. RelA integrates several environmental cues and synthesizes the alarmone ppGpp, which globally reprograms transcription, translation, and replication. We have developed and implemented novel single-molecule tracking methodology to characterize the intracellular catalytic cycle of RelA. Our single-molecule experiments show that RelA is on the ribosome under nonstarved conditions and that the individual enzyme molecule stays off the ribosome for an extended period of time after activation. This suggests that the catalytically active part of the RelA cycle is performed off, rather than on, the ribosome, and that rebinding to the ribosome is not necessary to trigger each ppGpp synthesis event. Furthermore, we find fast activation of RelA in response to heat stress followed by RelA rapidly being reset to its inactive state, which makes the system sensitive to new environmental cues and hints at an underlying excitable response mechanism.
(Less)
- author
- English, Brian P.
; Hauryliuk, Vasili
LU
; Sanamrad, Arash
; Tankov, Stoyan
; Dekker, Nynke H.
and Elf, Johan
- publishing date
- 2011-08-02
- type
- Contribution to journal
- publication status
- published
- keywords
- Cytosolic diffusion, Photoactivated localization microscopy, Single particle tracking, Stroboscopic illumination
- in
- Proceedings of the National Academy of Sciences of the United States of America
- volume
- 108
- issue
- 31
- pages
- 365 - 373
- publisher
- National Academy of Sciences
- external identifiers
-
- pmid:21730169
- scopus:79961224232
- ISSN
- 0027-8424
- DOI
- 10.1073/pnas.1102255108
- language
- English
- LU publication?
- no
- additional info
- Copyright: Copyright 2012 Elsevier B.V., All rights reserved.
- id
- c8783955-1b5c-4e54-89e5-8c15eab5854c
- date added to LUP
- 2021-09-24 20:47:14
- date last changed
- 2025-07-29 19:26:13
@article{c8783955-1b5c-4e54-89e5-8c15eab5854c,
abstract = {{<p>The RelA-mediated stringent response is at the heart of bacterial adaptation to starvation and stress, playing a major role in the bacterial cell cycle and virulence. RelA integrates several environmental cues and synthesizes the alarmone ppGpp, which globally reprograms transcription, translation, and replication. We have developed and implemented novel single-molecule tracking methodology to characterize the intracellular catalytic cycle of RelA. Our single-molecule experiments show that RelA is on the ribosome under nonstarved conditions and that the individual enzyme molecule stays off the ribosome for an extended period of time after activation. This suggests that the catalytically active part of the RelA cycle is performed off, rather than on, the ribosome, and that rebinding to the ribosome is not necessary to trigger each ppGpp synthesis event. Furthermore, we find fast activation of RelA in response to heat stress followed by RelA rapidly being reset to its inactive state, which makes the system sensitive to new environmental cues and hints at an underlying excitable response mechanism.</p>}},
author = {{English, Brian P. and Hauryliuk, Vasili and Sanamrad, Arash and Tankov, Stoyan and Dekker, Nynke H. and Elf, Johan}},
issn = {{0027-8424}},
keywords = {{Cytosolic diffusion; Photoactivated localization microscopy; Single particle tracking; Stroboscopic illumination}},
language = {{eng}},
month = {{08}},
number = {{31}},
pages = {{365--373}},
publisher = {{National Academy of Sciences}},
series = {{Proceedings of the National Academy of Sciences of the United States of America}},
title = {{Single-molecule investigations of the stringent response machinery in living bacterial cells}},
url = {{http://dx.doi.org/10.1073/pnas.1102255108}},
doi = {{10.1073/pnas.1102255108}},
volume = {{108}},
year = {{2011}},
}