Cargando…

The Escherichia coli S2P intramembrane protease RseP regulates ferric citrate uptake by cleaving the sigma factor regulator FecR

Escherichia coli RseP, a member of the site-2 protease family of intramembrane proteases, is involved in the activation of the σ(E) extracytoplasmic stress response and elimination of signal peptides from the cytoplasmic membrane. However, whether RseP has additional cellular functions is unclear. I...

Descripción completa

Detalles Bibliográficos
Autores principales: Yokoyama, Tatsuhiko, Niinae, Tomoya, Tsumagari, Kazuya, Imami, Koshi, Ishihama, Yasushi, Hizukuri, Yohei, Akiyama, Yoshinori
Formato: Online Artículo Texto
Lenguaje:English
Publicado: American Society for Biochemistry and Molecular Biology 2021
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC8144685/
https://www.ncbi.nlm.nih.gov/pubmed/33865858
http://dx.doi.org/10.1016/j.jbc.2021.100673
_version_ 1783697010585501696
author Yokoyama, Tatsuhiko
Niinae, Tomoya
Tsumagari, Kazuya
Imami, Koshi
Ishihama, Yasushi
Hizukuri, Yohei
Akiyama, Yoshinori
author_facet Yokoyama, Tatsuhiko
Niinae, Tomoya
Tsumagari, Kazuya
Imami, Koshi
Ishihama, Yasushi
Hizukuri, Yohei
Akiyama, Yoshinori
author_sort Yokoyama, Tatsuhiko
collection PubMed
description Escherichia coli RseP, a member of the site-2 protease family of intramembrane proteases, is involved in the activation of the σ(E) extracytoplasmic stress response and elimination of signal peptides from the cytoplasmic membrane. However, whether RseP has additional cellular functions is unclear. In this study, we used mass spectrometry–based quantitative proteomic analysis to search for new substrates that might reveal unknown physiological roles for RseP. Our data showed that the levels of several Fec system proteins encoded by the fecABCDE operon (fec operon) were significantly decreased in an RseP-deficient strain. The Fec system is responsible for the uptake of ferric citrate, and the transcription of the fec operon is controlled by FecI, an alternative sigma factor, and its regulator FecR, a single-pass transmembrane protein. Assays with a fec operon expression reporter demonstrated that the proteolytic activity of RseP is essential for the ferric citrate–dependent upregulation of the fec operon. Analysis using the FecR protein and FecR-derived model proteins showed that FecR undergoes sequential processing at the membrane and that RseP participates in the last step of this sequential processing to generate the N-terminal cytoplasmic fragment of FecR that participates in the transcription of the fec operon with FecI. A shortened FecR construct was not dependent on RseP for activation, confirming this cleavage step is the essential and sufficient role of RseP. Our study unveiled that E. coli RseP performs the intramembrane proteolysis of FecR, a novel physiological role that is essential for regulating iron uptake by the ferric citrate transport system.
format Online
Article
Text
id pubmed-8144685
institution National Center for Biotechnology Information
language English
publishDate 2021
publisher American Society for Biochemistry and Molecular Biology
record_format MEDLINE/PubMed
spelling pubmed-81446852021-05-26 The Escherichia coli S2P intramembrane protease RseP regulates ferric citrate uptake by cleaving the sigma factor regulator FecR Yokoyama, Tatsuhiko Niinae, Tomoya Tsumagari, Kazuya Imami, Koshi Ishihama, Yasushi Hizukuri, Yohei Akiyama, Yoshinori J Biol Chem Research Article Escherichia coli RseP, a member of the site-2 protease family of intramembrane proteases, is involved in the activation of the σ(E) extracytoplasmic stress response and elimination of signal peptides from the cytoplasmic membrane. However, whether RseP has additional cellular functions is unclear. In this study, we used mass spectrometry–based quantitative proteomic analysis to search for new substrates that might reveal unknown physiological roles for RseP. Our data showed that the levels of several Fec system proteins encoded by the fecABCDE operon (fec operon) were significantly decreased in an RseP-deficient strain. The Fec system is responsible for the uptake of ferric citrate, and the transcription of the fec operon is controlled by FecI, an alternative sigma factor, and its regulator FecR, a single-pass transmembrane protein. Assays with a fec operon expression reporter demonstrated that the proteolytic activity of RseP is essential for the ferric citrate–dependent upregulation of the fec operon. Analysis using the FecR protein and FecR-derived model proteins showed that FecR undergoes sequential processing at the membrane and that RseP participates in the last step of this sequential processing to generate the N-terminal cytoplasmic fragment of FecR that participates in the transcription of the fec operon with FecI. A shortened FecR construct was not dependent on RseP for activation, confirming this cleavage step is the essential and sufficient role of RseP. Our study unveiled that E. coli RseP performs the intramembrane proteolysis of FecR, a novel physiological role that is essential for regulating iron uptake by the ferric citrate transport system. American Society for Biochemistry and Molecular Biology 2021-04-16 /pmc/articles/PMC8144685/ /pubmed/33865858 http://dx.doi.org/10.1016/j.jbc.2021.100673 Text en © 2021 The Authors https://creativecommons.org/licenses/by/4.0/This is an open access article under the CC BY license (http://creativecommons.org/licenses/by/4.0/).
spellingShingle Research Article
Yokoyama, Tatsuhiko
Niinae, Tomoya
Tsumagari, Kazuya
Imami, Koshi
Ishihama, Yasushi
Hizukuri, Yohei
Akiyama, Yoshinori
The Escherichia coli S2P intramembrane protease RseP regulates ferric citrate uptake by cleaving the sigma factor regulator FecR
title The Escherichia coli S2P intramembrane protease RseP regulates ferric citrate uptake by cleaving the sigma factor regulator FecR
title_full The Escherichia coli S2P intramembrane protease RseP regulates ferric citrate uptake by cleaving the sigma factor regulator FecR
title_fullStr The Escherichia coli S2P intramembrane protease RseP regulates ferric citrate uptake by cleaving the sigma factor regulator FecR
title_full_unstemmed The Escherichia coli S2P intramembrane protease RseP regulates ferric citrate uptake by cleaving the sigma factor regulator FecR
title_short The Escherichia coli S2P intramembrane protease RseP regulates ferric citrate uptake by cleaving the sigma factor regulator FecR
title_sort escherichia coli s2p intramembrane protease rsep regulates ferric citrate uptake by cleaving the sigma factor regulator fecr
topic Research Article
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC8144685/
https://www.ncbi.nlm.nih.gov/pubmed/33865858
http://dx.doi.org/10.1016/j.jbc.2021.100673
work_keys_str_mv AT yokoyamatatsuhiko theescherichiacolis2pintramembraneproteasersepregulatesferriccitrateuptakebycleavingthesigmafactorregulatorfecr
AT niinaetomoya theescherichiacolis2pintramembraneproteasersepregulatesferriccitrateuptakebycleavingthesigmafactorregulatorfecr
AT tsumagarikazuya theescherichiacolis2pintramembraneproteasersepregulatesferriccitrateuptakebycleavingthesigmafactorregulatorfecr
AT imamikoshi theescherichiacolis2pintramembraneproteasersepregulatesferriccitrateuptakebycleavingthesigmafactorregulatorfecr
AT ishihamayasushi theescherichiacolis2pintramembraneproteasersepregulatesferriccitrateuptakebycleavingthesigmafactorregulatorfecr
AT hizukuriyohei theescherichiacolis2pintramembraneproteasersepregulatesferriccitrateuptakebycleavingthesigmafactorregulatorfecr
AT akiyamayoshinori theescherichiacolis2pintramembraneproteasersepregulatesferriccitrateuptakebycleavingthesigmafactorregulatorfecr
AT yokoyamatatsuhiko escherichiacolis2pintramembraneproteasersepregulatesferriccitrateuptakebycleavingthesigmafactorregulatorfecr
AT niinaetomoya escherichiacolis2pintramembraneproteasersepregulatesferriccitrateuptakebycleavingthesigmafactorregulatorfecr
AT tsumagarikazuya escherichiacolis2pintramembraneproteasersepregulatesferriccitrateuptakebycleavingthesigmafactorregulatorfecr
AT imamikoshi escherichiacolis2pintramembraneproteasersepregulatesferriccitrateuptakebycleavingthesigmafactorregulatorfecr
AT ishihamayasushi escherichiacolis2pintramembraneproteasersepregulatesferriccitrateuptakebycleavingthesigmafactorregulatorfecr
AT hizukuriyohei escherichiacolis2pintramembraneproteasersepregulatesferriccitrateuptakebycleavingthesigmafactorregulatorfecr
AT akiyamayoshinori escherichiacolis2pintramembraneproteasersepregulatesferriccitrateuptakebycleavingthesigmafactorregulatorfecr