Cargando…

Localization of EccA(3) at the growing pole in Mycobacterium smegmatis

BACKGROUND: Bacteria require specialized secretion systems for the export of molecules into the extracellular space to modify their environment and scavenge for nutrients. The ESX-3 secretion system is required by mycobacteria for iron homeostasis. The ESX-3 operon encodes for one cytoplasmic compon...

Descripción completa

Detalles Bibliográficos
Autores principales: Kriel, Nastassja L., Newton-Foot, Mae, Bennion, Owen T., Aldridge, Bree B., Mehaffy, Carolina, Belisle, John T., Walzl, Gerhard, Warren, Robin M., Sampson, Samantha L., Gey van Pittius, Nico C.
Formato: Online Artículo Texto
Lenguaje:English
Publicado: BioMed Central 2022
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC9118679/
https://www.ncbi.nlm.nih.gov/pubmed/35590245
http://dx.doi.org/10.1186/s12866-022-02554-6
_version_ 1784710549733376000
author Kriel, Nastassja L.
Newton-Foot, Mae
Bennion, Owen T.
Aldridge, Bree B.
Mehaffy, Carolina
Belisle, John T.
Walzl, Gerhard
Warren, Robin M.
Sampson, Samantha L.
Gey van Pittius, Nico C.
author_facet Kriel, Nastassja L.
Newton-Foot, Mae
Bennion, Owen T.
Aldridge, Bree B.
Mehaffy, Carolina
Belisle, John T.
Walzl, Gerhard
Warren, Robin M.
Sampson, Samantha L.
Gey van Pittius, Nico C.
author_sort Kriel, Nastassja L.
collection PubMed
description BACKGROUND: Bacteria require specialized secretion systems for the export of molecules into the extracellular space to modify their environment and scavenge for nutrients. The ESX-3 secretion system is required by mycobacteria for iron homeostasis. The ESX-3 operon encodes for one cytoplasmic component (EccA(3)) and five membrane components (EccB3 – EccE3 and MycP(3)). In this study we sought to identify the sub-cellular location of EccA(3) of the ESX-3 secretion system in mycobacteria. RESULTS: Fluorescently tagged EccA(3) localized to a single pole in the majority of Mycobacterium smegmatis cells and time-lapse fluorescent microscopy identified this pole as the growing pole. Deletion of ESX-3 did not prevent polar localization of fluorescently tagged EccA(3), suggesting that EccA(3) unipolar localization is independent of other ESX-3 components. Affinity purification - mass spectrometry was used to identify EccA(3) associated proteins which may contribute to the localization of EccA(3) at the growing pole. EccA(3) co-purified with fatty acid metabolism proteins (FAS, FadA3, KasA and KasB), mycolic acid synthesis proteins (UmaA, CmaA1), cell division proteins (FtsE and FtsZ), and cell shape and cell cycle proteins (MurS, CwsA and Wag31). Secretion system related proteins Ffh, SecA1, EccA1, and EspI were also identified. CONCLUSIONS: Time-lapse microscopy demonstrated that EccA3 is located at the growing pole in M. smegmatis. The co-purification of EccA(3) with proteins known to be required for polar growth, mycolic acid synthesis, the Sec secretion system (SecA1), and the signal recognition particle pathway (Ffh) also suggests that EccA(3) is located at the site of active cell growth. SUPPLEMENTARY INFORMATION: The online version contains supplementary material available at 10.1186/s12866-022-02554-6.
format Online
Article
Text
id pubmed-9118679
institution National Center for Biotechnology Information
language English
publishDate 2022
publisher BioMed Central
record_format MEDLINE/PubMed
spelling pubmed-91186792022-05-20 Localization of EccA(3) at the growing pole in Mycobacterium smegmatis Kriel, Nastassja L. Newton-Foot, Mae Bennion, Owen T. Aldridge, Bree B. Mehaffy, Carolina Belisle, John T. Walzl, Gerhard Warren, Robin M. Sampson, Samantha L. Gey van Pittius, Nico C. BMC Microbiol Research BACKGROUND: Bacteria require specialized secretion systems for the export of molecules into the extracellular space to modify their environment and scavenge for nutrients. The ESX-3 secretion system is required by mycobacteria for iron homeostasis. The ESX-3 operon encodes for one cytoplasmic component (EccA(3)) and five membrane components (EccB3 – EccE3 and MycP(3)). In this study we sought to identify the sub-cellular location of EccA(3) of the ESX-3 secretion system in mycobacteria. RESULTS: Fluorescently tagged EccA(3) localized to a single pole in the majority of Mycobacterium smegmatis cells and time-lapse fluorescent microscopy identified this pole as the growing pole. Deletion of ESX-3 did not prevent polar localization of fluorescently tagged EccA(3), suggesting that EccA(3) unipolar localization is independent of other ESX-3 components. Affinity purification - mass spectrometry was used to identify EccA(3) associated proteins which may contribute to the localization of EccA(3) at the growing pole. EccA(3) co-purified with fatty acid metabolism proteins (FAS, FadA3, KasA and KasB), mycolic acid synthesis proteins (UmaA, CmaA1), cell division proteins (FtsE and FtsZ), and cell shape and cell cycle proteins (MurS, CwsA and Wag31). Secretion system related proteins Ffh, SecA1, EccA1, and EspI were also identified. CONCLUSIONS: Time-lapse microscopy demonstrated that EccA3 is located at the growing pole in M. smegmatis. The co-purification of EccA(3) with proteins known to be required for polar growth, mycolic acid synthesis, the Sec secretion system (SecA1), and the signal recognition particle pathway (Ffh) also suggests that EccA(3) is located at the site of active cell growth. SUPPLEMENTARY INFORMATION: The online version contains supplementary material available at 10.1186/s12866-022-02554-6. BioMed Central 2022-05-19 /pmc/articles/PMC9118679/ /pubmed/35590245 http://dx.doi.org/10.1186/s12866-022-02554-6 Text en © The Author(s) 2022 https://creativecommons.org/licenses/by/4.0/Open AccessThis article is licensed under a Creative Commons Attribution 4.0 International License, which permits use, sharing, adaptation, distribution and reproduction in any medium or format, as long as you give appropriate credit to the original author(s) and the source, provide a link to the Creative Commons licence, and indicate if changes were made. The images or other third party material in this article are included in the article's Creative Commons licence, unless indicated otherwise in a credit line to the material. If material is not included in the article's Creative Commons licence and your intended use is not permitted by statutory regulation or exceeds the permitted use, you will need to obtain permission directly from the copyright holder. To view a copy of this licence, visit http://creativecommons.org/licenses/by/4.0/ (https://creativecommons.org/licenses/by/4.0/) . The Creative Commons Public Domain Dedication waiver (http://creativecommons.org/publicdomain/zero/1.0/ (https://creativecommons.org/publicdomain/zero/1.0/) ) applies to the data made available in this article, unless otherwise stated in a credit line to the data.
spellingShingle Research
Kriel, Nastassja L.
Newton-Foot, Mae
Bennion, Owen T.
Aldridge, Bree B.
Mehaffy, Carolina
Belisle, John T.
Walzl, Gerhard
Warren, Robin M.
Sampson, Samantha L.
Gey van Pittius, Nico C.
Localization of EccA(3) at the growing pole in Mycobacterium smegmatis
title Localization of EccA(3) at the growing pole in Mycobacterium smegmatis
title_full Localization of EccA(3) at the growing pole in Mycobacterium smegmatis
title_fullStr Localization of EccA(3) at the growing pole in Mycobacterium smegmatis
title_full_unstemmed Localization of EccA(3) at the growing pole in Mycobacterium smegmatis
title_short Localization of EccA(3) at the growing pole in Mycobacterium smegmatis
title_sort localization of ecca(3) at the growing pole in mycobacterium smegmatis
topic Research
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC9118679/
https://www.ncbi.nlm.nih.gov/pubmed/35590245
http://dx.doi.org/10.1186/s12866-022-02554-6
work_keys_str_mv AT krielnastassjal localizationofecca3atthegrowingpoleinmycobacteriumsmegmatis
AT newtonfootmae localizationofecca3atthegrowingpoleinmycobacteriumsmegmatis
AT bennionowent localizationofecca3atthegrowingpoleinmycobacteriumsmegmatis
AT aldridgebreeb localizationofecca3atthegrowingpoleinmycobacteriumsmegmatis
AT mehaffycarolina localizationofecca3atthegrowingpoleinmycobacteriumsmegmatis
AT belislejohnt localizationofecca3atthegrowingpoleinmycobacteriumsmegmatis
AT walzlgerhard localizationofecca3atthegrowingpoleinmycobacteriumsmegmatis
AT warrenrobinm localizationofecca3atthegrowingpoleinmycobacteriumsmegmatis
AT sampsonsamanthal localizationofecca3atthegrowingpoleinmycobacteriumsmegmatis
AT geyvanpittiusnicoc localizationofecca3atthegrowingpoleinmycobacteriumsmegmatis